GENETIC, PHYSICO-CHEMICAL AND STRUCTURAL
PARAMETERS AFFECTING TEXTURE OF DRY
EDIBLE BEANS
By
Agbo Nili Georges
AN ABSTRACT OF A DISSERTATION
Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of
DOCTOR OF PHILOSOPHY
Department of Food Science and Human Nutrition
1982

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TRADUCTION BREVE (en FRANÇAIS) DE LA THESE DE Ph.D.
PARAMETRES GENETIQUES, PHYSICO-CHIMIQUES ET STRUCTURAUX
AFFECTANT LA TEXTURE DES HARICOTS SECS COMESTIBLES
AGBO N'ZI GEORGES

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PARAMETRES GENETIQUES, PHYSICO-CHIMIQUES
ET STRUCTUR~UX AFFECTANT LA TEXTURE DES
HARICOTS SECS COMESTIBLES
RESUME
L'objectif de la présente étude a été d'étudier les carac-
téristiques physico-chimiques des graines de haricots secs en
relation avec la qualité de cuisson. Deux isolines (variétés
qui diffèrent génétiquement l'un de l'autre par seulement la
position d'un gène particulier sur un chromosome) de haricots
secs, San-Fernando (cultivar noir d'origine tropicale) et
Nep-2 (mutant blanc du San-Fernando) differ~nts seulement par
la couleur de la peau de leurs graines ont été comparés à raide
d'analyses approximatives, pour le comportement au trempage et
à la cuisson,
p:our les caractéristiques de leur farine. Sani-
lac, haricot sec (Phaseolus vulgaris L.) commercialement bien
connu a été utilisé comme standard pour les évaluations.
Les teneurs en humidité, protéines, amidon et cendres
n'ont pas montré de differences significatives entre Nep-2 et
San-Fernando. Par contre les résultats sur trois années de ré-
colte (1978, 1979 et 1980) ont montré des differences signifi-
catives seulement dans la teneur des lipides, Nep-2 ayant les
valeurs les plus élevées.
Oans l~ cotyledon, l~s te.n~ur~ en alph~-oligosaccharid~s
sont semblables dans les trois génotypes avec leur teneur en
stachyose plus importante. Aucun oligosaccharide n'a été dé-
tecté dans la peau chez les trois cultivars étudiés.
La vitesse d'absorption de l'eau est moins élevée avec· -
le San-Fernando qu'avec le Nep-2 et le Sanilac. Après cuisson,
les graines de sanilac et de Nep-2 présentent des quantités
plus élevées de liquide drainé et une texture plus faible par
rapport au San-Fernando. Aucune difference significative n'a
été détectée entre les cultivars en ce qui concerne l'humidi-
té après le trempage et la cuisson. La viscosité des farines
de ces haricots a indiqué des températures initiales de pâte
élevées avec des courbes croissantes continuelles de pâte pour
tous les cultivars utilisés. L'enlèvement de la peau des grai-
nes a augmenté le potentiel de viscosité des farines de Sani-
lac et San-Fernando. Cependant, la farine de Nep-2 a montré
une décroissance de l'allure de la courbe.
Une variabilité entre les génotypes a été observée au
microscope électronique à balayage pour les caractéristiques
structurales.' des graines. Nep-2 avait un micropyle partielle-
ment ouvert, la partie hypocotyle légèrement élevée et une peau
type élastique avec des trous alors que le San-Fernando avait
le micropyle complètement fermé, la partie hypocotyle élevée,
la peau rigide sans trous, l'organisation des cellules du pa-
renchyme' fermement entassées. dans les. graines ,et les grains

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d'amidon partiellement libérés dans les graines trempées et
cuites.

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INTRODUCTION
Les problèmes majeurs que rencontre le monde d'aujourd 'hui
sont de deux ordres:
- L'un concerne la croissance grandissante de la population
- L'autre la quantité insuffisante de provision alimentaire.
La médecine moderne et l'amélioration de l'hygiène chez
les hommes ont permis une augmentation spectaculaire de la du-
rée moyenne de vie de l'homme pendant que le taux de croissan-
ce dans beaucoup de pays reste pratiquement le même. Le résul-
tat est une explosion de la population dont parlent les écono-
mistes et autres spécialistes.
Les efforts fournis pour suivre le taux de naissance et
l'augmentation de l'approvionnement des produits vivriers sont
1
sévèrement limités par le manque d'éducation, l'attachement
aux coutumes, et la léthargie de la malnutrition et la sous-
nutrition.
Le problème de la croissance démographique et du manque
de nourriture n'est pas nouveau et tous les pays du monde en
ont eu des expériences depuis des millions d'années que l'hom-
me vit sur terre. Les problèmes présents diffèrent seulement
en ampleur et complexité de ceux qui ont fait rage auparavant.
Aujourd'hui, huit cent millions de gens de la population mon-_~
diale ne reçoivent pas une quantité adéquate de nourriture;
plus encore, environ la moitié de cette population mondiale
arrive à peine à se nourrir. Ce manque de nourriture à cette
catégorie de la population mondiale, principalement l'Afrique,
l'Amérique latine et l'Asie, peut être associé à plusieurs
facteurs tels que la sécheresse; l'infertilité des sols; la
rareté des terres cultivables (Sahel, régions Sud Saharienne);
la production inadéquate de produits vivriers due au fait que
le paysan produit pour nourrir sa petite famille et non pour
la commercialisation; le manque de moyens mécaniques pour de
larges productions; le manque de moyens de préservation, de
transformation et de transport, les limitations politiques et
économiques; les bas salaires aboutissant au manque de moyens
suffisants pour l'achat de nourriture de bonne qualité nutri-
tionnelle; le manque d'information sur la nutrition, l'amélio-
ration de la qualité et du rendement des produits disponibles.
Tous ces facteurs contribuent à la consommation de faible ca-
lorie par le groupe ci-dessus mentionné de la population mon-
diale.
En association avec le manque de nourriture il y a l'in-
suffisance de nutriments spécifiques nécessaires au bon fonc-
tionnement de l'organisme hum~in. Il y a six ~rincipales caté-
gories de nutriments qui sont nécessaires pour fournir de l'é-
nergie, promouvoir
la croissance, réparer les tissus et ré-
gulariser les processus métaboliques du co~ps. Ce sont: l'eau
les glucides, les protéines, les lipides, les vitamines et les
minéraux. Ces nutriments sont en général présents dans une

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large variété d'aliments mais limités dans d'autres. Par man-
que d'informations sur la nutrition et une distribution ina-
déquate, on peut être exposé à une insuffisance d'un de ces
nutriments pouvant entraîner différentes maladies.
Le nutriment qui a reçu une attention considérable
jusqu'à présent est la protéine.
La déficience en protéine animale dans le régime ali-
mentaire des régions dépourvues
du monde a généralement été
associée au développement physique des adultes et beaucoup
plus souvent aux maladies de déficience en protéine chez les
enfants.
Le Kwashiorkor (déficience en protéine, calorie) et
le marasme (ématiation due à la malnutrition) sont les deux
maladies qui causent une mortalité assez élevée parmi les en-
fants au moment du sevrage jusqu'à l'âge de six ans et souvent
les enfants qui survivent sont physiquement faibles ou sont
mentalement retardés.
Différentes autorités ont fait des propositionsen ce
qui concerne la composition optimale en protéine animale des
différents régimes alimentaires.
Le Département d'Agriculture des Etats-Unis avait pro-
posé 10 gr de protéine animale par personne par jour et en ce
qui concerne la protéine provenant des légumineuses ce taux
pourrait être atteint en augmentant la culture et l'utilisation
des légumineuses. Cook (FAO', 1962) dans son article "Population
and Food Supply" avait proposé d'atteindre 16 gr de protéine
animal par personne par jour pour les pays sous-alimentés.
Pawley (1963) dans son article "Possibility of Increasing
World Food Production" visant l'an 2000, prévoyait 20 gr de
protéine animale par personne par jour avec le reste des be-
soins alimentaires provenant d'autres sources.
Vue l'importance majeure d'une protéine complète (com-
me l'oeuf) dans le régime alimentaire particulièrement pour
le développement des enfants, il n'est pas facile de réaliser
ces objectifs.
En 1973, quatre raisons de la faible consommation de
protéine ont été mentionnées par le groupe Consultatif Calorie-
Protéine aux Nations Unies comme étant :
1. Salaires et revenus faibles et sous-emploi ou chômage dans
les centres ruraux et urbains, tout cela limite l'achat
d'aliments relativement chers qui contiennent de la protéi-
ne de qualité.
2. Difficultés associées à la production d'aliments riches en
protéine d'origine animale et végétale à cause des limita-
tions écologiques et agricoles résultant de la chèreté et
de la réduction de l'approvisionnement de ces produits.

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3. Le manque de connaissance des valeurs nutritives des ali-
ments et de leur préparation pour les enfantset des préju-
dices spécifiques privant ces enfants de la consommation
de certains aliments nécessaires (surtout quant ils sont
malades).
4. Le manque de traitement ou de transformation effective, le
manque d'un système adéquat de distribution et de marketing
entraînant une grande perte des semences de produits vivri-
ers.
Plusieurs approches ont été proposées pour résoudre ce
problème de protéine mais on doit reconnaître que la situation
est très complexe. Certains ont proposé la production autosuf-
fisante de céréales qui pourrait suppléer le manque de protéine
animale puis le développement de nouveaux produits alimentaires
riches en protéine. En 1968, le Comité Consultatif des Nations
Unies :a proposé :
1. La pro~otion de sources de protéine d'origine animale ou
végétales directement consommables par l'homme.
2. L'amélioration de l'efficacité
et de l'étendue des opérations
de pêche et autres produits marins.
3. La prévention des pertes inutiles d'aliments riches en
protéine pendant la conservation, le transport et la con-
sommation à domicile.
4. L'encouragement de l'utilisation directe d'aliments de
source oléagineuse riche en protéine pour l'homme.
5. Le promotion de la production et l'utilisation de sous-pro-
duits de pêche en protéine concentrée.
6. L'augmentation de la production et l'utilisation d'acides
animés synthétiques pour améliorer la qualité de la protéi-
ne dans les céréales et autres sources végétales puis le
développement de l'utilisation d'autres nutriments synthé-
tiques.
7. La promotion du développement de la protéine unicellulaire
pour l'alimentation des hommes et des animaux.
Un scientifique (Kahn 1981) indiqua qu'en 1985 les be-
soins alimentaires pour la population mondiale pourraient être
44 % plus élevés que ceux oonstatés en 1970 ; et environ 70 %
de cette est~mation de 1985 sera nécessaire pour nourrir la
population du Tiers-Monde. Son approche pour résoudre le pro-
blème est multisectorielle.
Il propose le contrOle de naissan-
ce, souhaita. le choix d'une stratégie par chaque gouvernement
pour extirper la pauvreté qui est associée à une faible con-·
sommation de protéine.
Il fit allusion que l'augmentation seu-
lement des revenus ne peut résoudre le problème de la pauvreté.
Sa dernière suggestion rejoignit celle d'autres seientifiques
pour l'amélioration du rendement de production de vivriers
dans plusieurs régions du monde malgré les différents facteurs
qui affectent la productivité.

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D: toutes les suggestions qui ont été présentées pour
la solut~on,du pro~lème de protéine, celle qui a reçu beau-
coup plus d attent~on et de support est l'amélioration de la
production et l'utilisation de protéine de sources animales
et végétales. Les produits de sources animales conviennent
naturellement pour la satisfaction des besoins en protéine
par l'homme à travers le monde entier, mais ils sont rares
chers, et quelquefois non utilisés à cause de l'ignorance,'
des tabous et la pauvreté.
La protéine de source végétale fournit plus de 80 % de
la protéine dans l'alimentation des pays en voie de dévelop-
pement. Les céréales, les légumineuses et les tubercules
donnent la plus grande contribution. La qualité de protéine
provenant des céréales et des tubercules n'est pas aussi
bonne que celle de source animale à cause de la limitation
de certains acides aminés essentiels tels que la lysine, le
tryptophane, la thréonine, la méthionine, la cystéine et l'his-
tidine.
A, cause de ce fait, une attention particulière fut prê-
tée aux légumineuses qui sont riches en lysine bien que pauvres
en acides aminés soufrés. Ceci les place comme compléments
naturels aux mêts à base de céréales et tubercules.
Carpenter (1981) avait signalé que la production mon-
diale de graines de légumineuses était en regression par rap-
port à celles de céréales à cause des hauts rendements obtenus
avec ces dernières. Cependant, les gouvernements et les agen-
ces internationales continuent d'encourager la production des
légumineuses à cause de leur capacité de fixation de l'azote
et à cause de l'effet indispensable dans l'alimentation des
gens qui digèrent mal les protéines d'origine laitière en par-
ticulier et d'origine animale en général.
Aux Etats-Unis, on remarque que le rendement de la pro-
duction des légumineuses est en baisse depuis 20 ans suggé-
rant-ainsi qu'un optimum a été déjà atteint dans le rendement
des variétés légumineuses qui étaient alors cultivées.
Au Michigan, à travers le programme "Amélioration
des
graines et grains du michigan", l'université d'Etat du Michi-
gan a été chargée du développement, de l'évaluation et de la
relance de nouvelles variétés. En effet, Hosfield et Uebersax
(1980) ont souligné le fait qu'il y a de nombreuses raisons
qui peuvent expliquer l'existence d'un top-niveau dans la pro-
duction des produits agricoles, et que toutes ces raisons ont
pour dénominateur commun la spécificité variétale. La variété
possède,selon ces auteurs, un certain potentiel génétique (ou
programme prescrit par ses constituents génétiques) qui lui
permet de produire sous un ensemble donné de conditions. Si
ces conditions sont optimales, la variété sera produite à son
rendement maximal; si les conditions changent et que celles
qui prévalent limitent le potentiel génétique de la variété,
son rendement sera faible. Cette variété doit alors être rem-

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5
placée par une autre mieux adaptée aux nouvelles conditions ef
qui peut exprimer son potentiel génétique en terme de produc-
tivité et de qualité. Frazier (1978) a indiqué que certains
hauts rendements obtenus dans la production de légumineuses
pendant les nombreux essais au Michigan sont obtenus avec les
haricots d'origine tropicale. Smucker, ~~. (1978) précisent
que les résultats préliminaires de plusieurs expériences sur
le champ ont montré que les haricots noirs d'origine tropicale
ont des systèmes de racines plus appropriées à la compacité
partielle du sol et aux contraintes de l'eau.
Les sous-familles:Minosaceae, caesielpinaceae et papi-
lionaceae de la famille léguminaseae sont souvent utilisées
pour l'alimentation. La sous-famille papilionaceae qui pousse
dans les régions tropicales et tempérées fournit la plupart .
des haricots comestibles. Dans cette thèse, l'espèce phaseolus
vulgaris h.de la sous-famille papilionaceae, haricot bien con-
nu et consommé en grande quantité dans le monde, sert d'exem-
ple pour l'examen des problèmes associés aux légumineuses. A
cause de la haute valeur nutritionnelle des haricots, plusieurs
chercheurs ont effectué des analyses quantitatives et qualita-
tives concernant les protéines, les lipides, l'humidité, les
fibres,
les cendres, les sels minéraux, les vitamines, les.
glucides, et la texture.
Il faut souvent une longue période de cuisson pour
faire cuir les haricots secs. L'hydratation des haricots avant
la cuisson est un facteur important. Des méthodes convention-
nelles de préparation et de cuisson des haricots secs compor-
tent le trempage toute une nuit dans l'eau dans les conditions
normales de températures suivi de cuisson pendant 45 minutes
à 3 heures.
Le temps de cuisson dépend de la variété et des
conditions de conservation. Plusieurs tests effectués à cet
effet ont montré que la connaissance d'un certain nombre de
facteurs tels que la composition chimique des tissus spécifi-
ques et la localisation des constituents chimiques dans ces
tissus était un préalable nécessaire à la comphréhension des
variations physico-chimiques dans les tissus des haricots pen-
dant leurs traitements mécanique, thermique, chimique et enzy-
matique (Powrie ~ ~l. 1960).
La texture du haricot cuit est une caractéristique
importante d'autant plus que son appréciation par le consomma-
teur est un facteur important pour la promotion d'un~_produc­
tian intensive. Bien qu'il y ait eu de nombreux essais pour
mesurer la texture des haricots cuits, on sait très peu de
chose sur la texture des haricots secs et trempés.
La consommation des haricots a été rendue responsable
de la production excessive de gaz dans le tube intestinal,
condition connue sous le' nom-de flatulence. Des études ont été
-effectuées pour essayer de conna1tre les facteurs de flatulen-
ce dans les haricots. Plusieurs sucres indigestibles tels que
le stachyose et le raffinose ont été ainsi identifiés comme
étant associés à la flatulence due à la consommation de haricot.

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6
Pour étudier ce problème, les recherches ont été axées princi-
palement sur les points suivan~ :
1. Caractérisation des composés spécifiques
2. Développement des moyens physiques et chimiques d'extrac-
tion de ces composés
3. Développement de techniques pratiques de traitement et de
cuisson pouvant réduire le phénomène de flatulence
4. Essais d'identification des variants génétiques possédant
de faibles teneurs en oligosaccharides et peuvent être
utilisés dans la consommation
Bien que le rendement soit le plus important critère
dans la promotion et le développement de la culture d'une nou-
velle variété de haricots, le généticien est toujours conscient
des habitudes culinaires du consommateur quand il introduit de
nouveaux cultivars. Les consommateurs ont acquis des habitudes
spécifiques qui prennent en compte des caractéristiques phéno-
typiques des haricots
)des conditions de trempage et de cuis-
son et,de la qualité nutritionnelle des protéines constituti-
ves (Rockland et Jones, 1974).
L'utilisation de technologies nouvelles a fourni des'
haricots avec lesquels les évaluations qualitatives peuvent
être conduites avec un maximum d'objectivité et de~précision'
sur de petites quantités de graines (HuIse etat 1977). Cependant,
on a peu d' i n for mat ion s sur les fa c t e urs co nau ù:; an t à l a var i a -
bilité des caractéristiques physico-chimiques observées dans
les haricots secs comestibles (Hosfield and Uebersax, 1980).
Les variétés qui diffèrent génétiquement l'un de l'au-
tre par seulement la position d'un gène particulier sur un
chromosome sont dénommées isolines. Nep-2 est le résultat
i
d'une mutation induite dans le San-Fernando qui probablement
a affecté le gène de c~uleur dans le haricot (Moh, 1971). Les
differences observées dans les propriétées physico-chimiques
1
de San-Fernando et Nep-2 étaient en conséquence inattendues et
f
motivèrent cette étude à cause de leur impact possible sur la
qualité de la cuisson. Des essais préliminaires de trempage de
{
ces cultivars ont montré que San-Fernando et Nep-2 diffèrent
{
par la vitesse d'absorption d'eau pendant le trempage. On a
remarqué aussi qu'il fallait deux fois plus de force pour
trancher un échantillon de San-Fernando que pour un échantil-
1
lon de Nep-2.
L'objectif de cette recherche a été d'étudier les dif-
.ferences physico-chimiques observées entre les deux isolines
mentionnés ci-dessus. Le cultivar haricot sec, Sanilac, a été
utilisé comme réference (standard) pour toutes les évaluations.
A cet effet, des analyses ont été effectuées pour avoir des
renseignements précis sur,
1. Ua- composition approximative des deux isolines et sanilac
( haricot standard commerciallement connu, Phaseolus
vulgaris L.)

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2. Les sucres de flatulence en vue de promouvoir la culture
des variétés génétiques qui en sont pauvres
3. La microstructure des trois cultivars par le microscope
à balayage afin d'établir les relations possibles entre
la texture et les differences physico-chimiques
4. Les vitesses d'hydratation des trois cultivars et leurs
relations avec leurs microstructures
5. Les propriétés physiques (viscosité) de farines de hari-
cots et leurs relations avec leurs caractéristiques de
pAte
6. La qualité de traitement et la texture des trois culti-
vars
7. L'effet du trempage et de cuisson sur la microstructure
des haricots et sa relation avec la texture
METHODES
Des méthodes et techniques appropriées ont été utili-
sées pour l'accomplissement des expériences ci-dessus mention-
nées (voir thèse).
Les échantillons ont été obtenus du champ expérimental
près de Saginaw vallée, avec le concours du département des
Sciences agricoles et du sol, Université d'Etat du Michigan,
East-Lansing, Michigan, pour trois années de récoltes.
RESULTATS
Les résultats présentés dans cette thèse sur l'analyse
approximative; la microstructure des haricots secs, trempés et
cuits; la texture et autres paramètres d'évaluation, sur le
produit cuit de sanilac, Nep-2 et San-Fernando ont fourni des
informations importantes et les conclusions importantes ont pu
être tirées comme suit:
L'analyse approximative d'échantillons sur trois années
de récoltes ont revelé qu'il n'y avait pas de differences si-
gnificatives entre
les cultivars étudiés à l'exception de
la difference dans la teneur en lipides dans les haricots des
années de récoltes 1979 et 1980.
L'analyse des sucres a montré des teneurs élevées
d'hexoses et de stachyose dans tous les trois cultivars. Le
raffinose et le stachyose n'ont été détectés dans les peaux_
d'aucun des cultivars.
L'étude de l'absorption d'eau a révelé que San-Fernan-
do a la plus basse courbe de vitesse d'absorption d'eau alors
que la teneur en protéine, qui est la fraction en majeur par-

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tie responsable de l'absorption d'eau d'un haricot n'a pas ré~
velé de differences significatives d'une variété à l'autre.
Cette réduction dans l'absorption d'eau par le San-Fernando a
été attribuée aux differences structurelles de la peau des
graines.
L'étude au microscope électronique à balayage sur les
graines sèches des trois cultivars a montré des structures
1
hautement organisées, communément trouvées dans les graines de
î
légumineuses. Les differentes structures anatomiques observées
pouvaient bien être utilisées pour expliquer certaines diffe-
t
rences de comportement dans les propriétés fonctionnelles tel-
1
les que l'absorption d'eau et la texture qui existent entrê les
deux isolines, Nep-2 et San-Fernando. Ces études ont conduit
1
à la conclusion que le quotient de 11 épaisseur du péricarpe etl.du té-
9ument de la peau de San-Fernando, les surfaces internes et
.
externes de la peau et du cotyledon ont tous un effet ~ur la
vitesse d'absorption de l'eau. La partie du micropyle et de
l'hypocotyle peut-être utilisée comme indice d'identification
des trois cultivars à cause de leur spécificité pour chaque
cultivar. Les graines de San-Fernando possèdent des ouvertures
caractéristiques du micropyle pendant le trempage; ce qui con-
tribut à son efficacité d'absorption d'eau pendant la cuisson
pour la préparation de conserves. Les cellules non brisées du
parenchyme observées dans les cotyledons de San-Fernando trem~
pés sont une indication de résistance accrue au tranchage dans
ce cultivar comparé au Sanilac qui présente une libération
complète des grains d'amidon de ces cellules. En outre, la
peau encore rigide et les grains d'amidon partiellement intacts
dans les graines cuites de San-Fernando contribuent à l'ac~
croissement de la résistance au tranchage des graines par Kra-
mer Shear Press. Ces observations distinctes pourraient aider
les généticiens agronomes dans leur strategie de sélection des
haricots.
La clarté de couleur des graines (valeur L) décroît
pendant la cuisson alors que la rougeur (valeur +a L) et le jau-
nissement (valeur +b L ) s'intensifient à l'exception des graines
du San-Fernando où il y avait une fluctuation dans les valeurs
d'année en année. Ceci est un processus prévisible qui généra-
lement se passe sous l'effet de la chaleur causant un brunis-
sement.
Il nly a pas de differences significatives entre les
cultivars en ce qui concerne la teneur en humidité après trem-
page et cuisson. Le produit cuit a révelé un haut poids de
drainage et une faible texture pour le sanilac et Nep-2 mais
l'inverse pour le San-Fernando.
Les differences observées en texture après l'analyse
par le Kramer Shear Press, entre les d~ux isolines pourraient
être attribuées aux differences structurelles obtenues qui'
existent au niveau de l'arrangement des cellules dans leur peau
'1'
et le parenchyme. Il est demandé beaucoup plus de temps pour
cuir les graines de San-Fernando que Nep-2 selon l'étude de la
texture sur les graines prises individuellement.

---

9
L'expérience sur la viscosité a montré une températu~
re initiale élevée de la pâte avec des courbes de type C pour
tous les cultivars. L'enlèvement de la peau de la graine
accroit le potentiel de viscosité du sanilac et San-Fernando
contrairement au Nep-2 qui a montré un viscosité décroissante.
La contribution des graines à la viscosité varie donc avec le
cultivar et montre une difference entre les deux isolines.
CONCLUSION .
Ce travail a fourni des résultats fondamentaux sur
les aspects génétiques et les caractéristiques physico-chimi-
t
ques et structurelles des haricots secs utilisés. Ces résul-
!
tats peuvent avoir une importance significative dans les ten-
t
tatives d'élaboration des changements génétiques (biologiques)
f
et technologiques (physiques) dans le but d'améliorer la quali-
1
té des haricots secs comestibles. Les efforts de recherche en
t
collaboration entre les généticiens agronomes et les spécia-
listes en Science de l'alimentation pourraient contribuer à la
réduction de l'énergie demandée pour préparer les haricots secs
pour la consommation et à l'amélioration de la cuisson et de la
qualité nutritionnelle des haricots cuits.
En général, la solution au problème de protéine est
une production suffisante d'aliments riches en protéines dans
les pays du Tiers monde. En Afrique où 150 millions de person-.
nes sont en danger de mort et où 500 000 tonnes d'aliments
font actuellement défaut pour nourrir les pays se trouvant dans
le besoin, nous avons besoin de prêter une attention particu~
lière à la production et 'à la quali té de nos denrées pour résou-
dre ce problème de faim qui affecte non seulement les personnes
mais aussi l'économie d'un pays. Nos denrées déjà utilisées
doivent être améliorées en rendement et qualité en effectuant
des études sur toutes ces denrées par l'application de certai-
nes techniques adaptables pour la résolution de ce problème de
notre temps. Plusieurs plantes indigènes aussi chez nous res-
tent encore inconnues- des pays industrialisés et il est néces-
saire de les étudier pour évaluer leurs aspects nutritionnels,
,--
toxicologiques et technologiques en vue de leur incorporation
-
dans le système agricole déjà en exploitation.
i
Ma thèse est un exemple d'étude qui peut très bien
s'adapter dans les priorités de programmes de développement
sur les légûmineuses ou autres denrées en Côte d'Ivoireet en
Afrique.

---

~·<·:/1
1
ABSTRACT
GENETIC, PHYSICO-CHEMICAL AND STRUCTURAL
PARAMETERS AFFECTING TEXTURE OF DRY
EDIBLE BEANS
By
Agbo N'li Georges
The purpo~e of the present study was to develop funda-
mental information concerning physical and chemical
characteristics of dry bean (Phaseolus vulgaris L.) seeds
and relate them to cooking quality.
Two dry bean isolines
San-Fernando (a black seeded cultivar of tropical origin)
and Nep-2 (a white seeded mutant from San-Fernando) presuma-
bly differing only in seed coat color were compared for
proximate analysis, soaking and cooking, and flour chàrac:
teristics.
Sanilac, a navy bean cultivar used widely in
dry bean studies was employed as a control.
Proximate composition of Nep-2 and San-Fernando showed
no significant differences for moisture, protein, fat,
starch and ash contents.
Cumulative proximate composition
over the three crop years (1978, 1979 and 1980) showed
sfgnif1cant d1fferences in fat contents only and Nep-2 had
higher fat values than San-Fernando.
In the cotyledon, a-oligosaccharide contents were
similar in all "three genotypes with stachyose being in the
greatest quanttty.
The oligosaccharides were not detected
in the seed coat 'of any of the three cultivars used,

---

To
my Mother
For her patience and moral support
,•
i i

---

ACKNOWLEDGEMENTS
1 wish to express my gratitude to my major professor,
Dr. M. Uebersax, for his guidance and patience during this
study and preparation of this dissertation.
1 address my
sincere thanks to Dr. G. Hosfield of the Department of Crops
and Soil Science, and his wife Marie, for their particular
concern and moral support in the accomplishment of this
work.
1 extend my appreciation to Dr. P.Markakis, Dr. L.
Dawson, and Dr. 1. Gray of the Department of Food Science
and Human Nutrition, and Dr. R. Herner of the Department of
Horticulture, for serving on my graduate committee with
their suggestions and reviewing of this manuscript.
1 express my deep sense of appreciation and love to my
. dear mother and late father Mrs. and Mr. Agbo, for their
wise advice and encouragement in many ways.
1 also feel grateful to the Ministry of Research,
Republic of Ivory Coast for the opportunity and financial
assistance granted to me and USDA program associated with
the Department of Crop and Soil Sciences at Michigan State
University for supplying the materials and equipment used.
1 further thank my cousins, Mr. N'li Bruno and Mr. Nili
Remy, and my friends, Mr. Iloki Ignace and Mr. Kraidi
Nicolas for their help when 1 needed it.
i i ;

---

TABLE OF CONTENTS
Page
LIST OF TABLES.
vii
LIST OF FIGURES .
x
INTRODUCTION . .
1
The Wor1d Food Prob1em.
. . . . . • • . • . . • .
3
"Protein Prob1em" . . .
. . .
. . . . • •
Possible Solutions to the "Protein Prob1em" . . .
4
Needs of Vegetab1e Proteins . . . . • . . . .
7
Yie1d Prob1em.
. . . . . • . . • . . • • . .
8
Bean Production and Uti1ization . . . . . . .
9
Objectives. .
. . . .
. • . . • .
13
LITERATURE REVIEW . .
1 5
Nutrition Contribution of Food Legume Seeds to
Wor1d Hunger. . . . .
. . . , . .
15
Cooking and Processing. . . . . . . . • .
17
Moisture Factor. . .
. . •
• . .
20
Hydration . . . : . .
. . . . • • . . . •
21
Texture Measurement Means . • • • • . . . . .
25
Re1ationship of Texture Hard-to-cook Beans..
28
Scanning Electron Microscopy (SEM).
. . . . . . .
31
Legume Seed Anatomy . . . . . . . . • . • . . . .
39
Starch and Viscosity. . . . . . . . . . . . . . .
43
Indigestib1e Bean Sugars Prob1em.
. . . . . .
50
Qua1ity Evaluation of Processed Products.
. . . .
57
MATERIALS AND METHODS . . .
61
Samp1e Source
.
. . . .
61
Seed Coat Separation.
61
Grinding of Samp1e . . . . .
. .
62
Moisture.
62
Crude Fat . .
. . .
62
As h . . . . .
62
Protein . . .
62
Sugars.
. . . . .
. . . . . . . . .
63
Extraction and Injection • . • .
63
Methods of Quantitation . .
. .
65
i v

---

Page
Starch
.
66
Sample Preparation . . . . . • . . . . . . . .
66
Buffer and Enzyme Solution
.
68
0.2 M acetate buffer stock solution
.
68
0.02 M acetate buffer working solution . .
68
Enzyme solutions. . .
. . . .
. . .
69
Enzyme Characteristics. . . . . . . .
• .
69
Hydrolysis. . . . . . .
. . .
. . . . .
69
Injection. . . . . . .
. . .
70
Methods of Quantitation .
. . .
70
Calculations.
. . . . . . . . . . .
. . .
70
Water Absorption in Beans .
. • . . . . .
71
Canning
.
71
Texture Measurement Based on Single Bean.
76
Gelatinization Characteristics • . . • . . • • . .
77
The peak viscosity or pasting peak . . . • . .
77
The viscosity at the end of the heating cycle
as sample reaches 95 0 C. . . . • . . . . . .
79
The viscosity at the end of the 95 0 C holding
cyc l e .
. .
.
.
. .
.
.
.
.
. ...
. .
.
.
.
79
The viscosity at end of the cooling cycle
when the paste ~eaches again 50 0 C • . . • .
79
The viscosity at the end of the 50 0 C holding
cyc l e .
.
.
.
.
. .
.
.
.
.
.
.
80
Scanning Electron Microscopy . .
81
Dehydration . . . .
81
Coating and viewing
81
Statistical Analysis.
82
RESULTS AND DISCUSSION . .
83
Proximate Composition
.
83
Sugars.
.
.
.
.
. .
. .
.
.
.
.
.
95
Scanning Electron Microscopy Examination of Dry
Beans
.
. . .
.
.
.
.
. .
. .
.
98
Water Absorption on Soaking . . . • . . •
116
Pasting Properties. . . .
. • .
120
SEM of Soaked Beans . . . .
. ••
129
SEM of Processed Beans. . .
. . . • • . .
134
Bean Co l or.
.
.
.
.
.
. .
.
.
.
.
.
. .
. .
138
Processed Bean Evaluation.
.
.
141
Moisture content. .
.
.
141
Drained weight . .
145
Clump and split. . . . . . . . • . .
••
146
Texture .
.
146
CONCLUSIONS . . .
. . . .
153
RECOMMENDATIONS FOR FUTURE RESEARCH • . . . . . .
157
v

---

Page
LITERATURE CITED . .
159
•
t
•
•
APPENDIX • • • • • .
. . 178
v;

---

LIST OF TABLES
Table
Page
1
Comparison between microscopes.
36
2
Legume testa feature characteristics . .
42
3
Bean processing evaluation and calculations .
74
4
Functional and molecular properties associated
with pasting characteristics of starches . . . . .
78
5
Percent moisture content (dry basis) of bean
cultivars (Sanilac, Nep-2 and San-Fernando) by
crop years (1978, 1979 and 1980) and seed por-
tions (whole bean, cotyledon and seed coat)
.
84
6
Percent moisture content (dry basis) of bean
cultivars (Sanilac, Nep-2 and San-Fernando)
for three crop years (1978, 1979 and 1980) by
seed portions (whole bean, cotyledon and seed
coat)
.
85
7
Percent protein content (dry basis) of.bean
cultivars (Sanilac, Nep-2 and San-Fernandl) by
crop years (1978, 1979 and 1980) and seed
portions (whole bean, cotyledon and seed coat) . .
86
8
Percent protein content (dry basis) of bean
cultivars (Sanilac, Nep-2 and San-Fernando) for
three crop years (1978, 1979 and 1980) by seed
portions (whole bean, cotyledon and seed coat) . .
87
9
Percent fat content (dry basis) of bean cultivars
(Sanilac, Nep-2 and San-Fernando) by crop years
(1978, 1979 and 1980) and seed portions (whole
bean, cotyledon and seed coat) . . . . • . . • . .
88
10
Percent fat content (dry basis) of bean cultivars
(Sanilac, Nep-2 and San-Fernando) for three crop
years (1978, 1979 and 1980) by seed portions
(whole bean, cotyledon and seed coat) . • • • • •
89
vii

---

Table
Page
11
Percent ash content (dry basis) of bean cultivars
(Sanilac, Nep-2 and San-Fernando) by crop years
(1978, 1979 and 1980) and seed portions (whole
bean, cotyledon and seed coat). . . . . . . . . .
90
12
Percent ash content (dry basis) of bean cultivars
(Sanilac, Nep-2 and San-Fernando) for three crop
years (1978, 1979 and 1980) by seed portions
(whole bean, cotyledon and seed coat)
. . . . . .
91
13
Percent starch content (dry ba~is) of bean culti-
vars (Sanilac, Nep-2 and San-Fernando) by crop
years (1978, 1979 and 1980) and seed portions
(whole bean, cotyledon and seed coat) . . . . . .
92
14
Percent starch content (dry basis) of bean culti-
vars (Sanilac, Nep-2 and San-Fernando) for three
crop years (1978, 1979 and 1980) by seed portions
(whole bean, cotyledon and seed coat) . . . . . .
93
15
Pe r ce nt s uga r (h ex0 se, suc r 0 se, i nos i t 0 l, ra f -
finose and stachyose) content (dry basis) of
bean ;cultivars (Sanilac, Nep-2 and San-Fernando)
and seed portions (whole bean, cotyledon and seed
coat)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
. .
96
16
Percent water absorption of dry beans soaked in
ambient temperature water (1:1, tap water:
distilled water) for up to 90 minutes by crop
years (1978, 1979 and 1980) . . . . . • . . . . .
117
17
Pasting characteristics of dry bean portions
(whole bean and cotyledon) flours by crop years
(1978, 1979 and 1980) and cultivars (Sanilac,
Nep-2 and San-Fernando) in a 400 ml phosphate
buffer solution at pH 5.30. . . . . . . . . . . .
121
18
Hunter lab color and color difference values (l,
al, bl) of dry and processed bean cultivars
(Sanilac, Nep-2 and San-Fernando) by crop years
(1978, 1979 and 1980) . . . . . . . . . . . . . .
140
19
Quality parameter Evaluation for processed bean.
cultivars (Sanilac, Nep-2 and San-Fernando) by
crop years (1978, 1979 and 1980)
143
20
Texture based on single bean cooking time
(minutes) for bean cultivars (Sanilac, Nep-2 and
San-Fernando) . . . . . . . . . . . . . . . . . .
152
vii i

---

Table
Page
lA
Percent sugars (hexose, sucrose, inositol, raf-
finose and stachyose) content (dry basis) of
bean cultivars (Sanilac, Nep-2 and San-Fernando)
for three crop years (1978, 1979 and 1980) by
seed portions (whole bean, cotyledon and seed
coat).
.
.
.
.
.
.
.
.
.
.
.
.
. .
.
.
.
.
.
. .
, 78
lB
Pasting characteristics of dry bean portions
(whole bean and cotyledon) flours for three crop
years (1978, 1979 and 1980) by cultivars (Sani-
lac, Nep-2 and San-Fernando) in a 400 ml
phosphate buffer solution at pH 5.30 . . • .
182
le
Hunter Lab color and color difference values of
dry and processed bean cultivars (Sanilac,
Nep-2 and San-Fernando) over three crop years
(1978, 1979 and 1980). . . . • . . • . • • •
186
lD
Processed bean evaluation - three crop years -
(1978, 1979 and 1980) bean samples . • . . • • .
190
i x

---

LIST OF FIGURES
Figure
Page
1
Phylogenetic relationship of several dry edible
legumes...
.
10
2
The principle of scanning electron microscopy..
32
3
Essential features of SEM . .
34
4
Visual observation of legume seed structure.
41
5
Structure relationships of the raffinose family
oligosaccharides . . . . . . .
. . .
53
6
Structure of galactopinitol . . .
. . . . . .
54
7
Flow diagram of sugar analysis .
64
8
Flow diagram of starch dispersion, hydrolysis
and analysis . . . . . . . . . . . . . . . • . .
67
9
Schematic diagram illustrating unit operations
for dry bean processing procedures
. . .
73
10
Flow diagram of sample preparation for SEM. . .
82
11
Mean sugar (hexose, sucrase, inositol, raffinose
and stachyose) content values over three crop
years (1978, 1979 and 1980) for whole bean
cultivars (Sanilac, Nep-2 and San-Fernando). . .
99
12
Mean sugar (hexose, sucrase, inositol, raffinose
and stachyose) content values over three crop
years (1978, 1979 and 1980) for cultivars
(Sanilac, Nep-2 and San-Fernando) cotyledons..
100
13
Mean sugar (hexose, sucrase, inositol, raffinose
and stachyose) content values over three crop
years (1978, 1979 and 1980) for bean cultivars
(Sanilac, Nep-2 and San-Fernando) seed coats . •
101
14
Scanning electron micrographs of dry bean seed
coat outer (lOOOx) and inner (lOOOx) surface
structures: Al = Sanilac coat outer surface;
x

---

Figure
Page
A2 = Sanilac inner surface; Bl = Nep-2 coat outer
surface; B2 - Nep-2 coat inner surface; Cl =
San-Fernando coat outer surface; C2 = San-Fer-
nando coat inner surface. • . . . . . . . • . • .
102
15
Scanning electron micrographs of dry bean seed
cotyledon outer surface structures (30x and
700x); Al and A2 = Sanilac cotyledon outer sur-
face; Bl and B2 = Nep-2 cotyledon outer surface;
Cl and C2 = San-Fernando outer surface . . • . . .
104
16
Scanning electron micrographs of dry bean seed
hilum area structures Al = Sanilac hilum area
(50x); A2 = Sanilac micropyle (lOOOx); Bl =
Nep - 2 hil um a rea (5 0x ); B2 = Nep - 2 mi c r 0 py l e
(lOOOx); Cl = San-Fernando hilum area (50x);
C2 = San-Fernando micropyle (lOOOx); HC = hypo-
cotyl area (50x); H = hilum; M = micropyle • . . •
106
17
Scanning electron micrographs of dry ·bean seed
hypocotyl area (lOOOx) and hilum transverse
cross-section (200x) structures Al = Sanilac
hypocotyl area; A2 = Sanilac hilum transverse
section; Bl = Nep-2 hypocotyl area; B2 = Nep-2
hilum transverse section; Cl = San-Fernando
hypocotyl area; C2 = San-Fernando hilum trans-
verse sec t ion . . • . . . . . . • . • • . . • • .
107
18
Scanning electron micrographs of dry bean seed
coat cross-section structures (1600x)
Al = Sanilac longitudinal cross-section;
A2 = Sanilac transverse cross-section;
Bl = Nep-2 longitudinal cross-section;
B2 = Nep-2 transverse cross-section
Cl = San-Fernando longitudinal cross-section
C2 = San-Fernando transverse section • . • • •
108
19
Scanning electron micrographs of dry bean seed
cotyledon inner side surface structures (30x and
400x).
Al and A2 = Sanilac cotyledon inner side
surface; Bl and 62 = Nep-2 cotyledon inner side
surface; Cl and C2 = San-Fernando cotyledon inner
si de surface. • • • . . • . • • . • • . • • . • •
110
20
Scanning electron micrographs of 1980 dry Nep-2
bean cotyledon outer and inner side surface
structures (30x); A = cotyledon outer side
surface; Band C = cotyledon inner side surface
112
x i

---

Figure
Page
21
Scanning e1ectron micrographs of dry bean seed
cotyledon cross-section structures (1600x).
Al = Sani1ac longitudinal cross-section; A2 =
Sani1ac transverse cross-section; B1 = Nep-2
longitudinal cross-section; B2 = Nep-2 trans-
verse cross-section; Cl = San-Fernando cross-
section; C2 = San-Fernando transverse cross-
section
.
113
22
Scanning e1ectron micrographs of dry bean seed
cotyledon cross-section structures at lower
magnification (400x).
Al = Sanilac longitudinal
section; A2 = Sani1ac transverse section; B1 =
Nep-2 longltudina1 section; B2 = Nep-2 trans-
verse section; Cl = San-Fernando longitudinal
section; C
115
2 = San-Fernando transverse section ••
23
Mean percent water absorption pattern of dry
bean cultivars (Sani1ac, Nep-2 and San-Fernando)
soaked in arnbient ternperature water (1:1, tap
water:disti11ed water) for up to 90 minutes over
three crop years (1978, 1979 and 1980) . . . . •
118
24
Mean pasting curves of dry bean f10urs (who1e
bean and cotyledon) by cultivars (Sani1ac,
Nep-2 and San-Fennando) over three crop years
(1978,1979 and 1980) . . . . . • . . . . . . . .
125
25
Mean pasting curves of dry Sani1ac bean f10urs
(who1e bean and cotyledon) over three crop years
(1978,1979 and 1980) . . . . . . . • . . . •
126
26
Mean pasting curves of dry Nep-2 bean f10urs
(who1e bean and cotyledon) over three crop years
(1978,1979 and 1980) . . . • . . . . . . . . . .
127
27
Mean pasting curves of dry San-Fernando f10urs
(who1e bean and cotyledon) over three crop years
(1978,1979 and 1980) . • • . . . . . • . . •
128
28
Scanning e1ectron micrographs of soaked bean
seed coat outer surface (1000x) and cross~sec~
tion (1000x) structures
Al = Sani1ac seed coat
outer surface structure; A2 = Sani1ac seed coat
cross-section structure; B1 = Nep-2 seed coat
outer surface structure; B2 = Nep-2 seed coat
cross~section structure; Cl = San-Fernando
xii

---

Figure
Page
seed coat outer surface structure; C2 = San-
Fernando seed coat cross-section structure • . . . 130
29
scanning electron micrographs of soaked bean
seed hilum area structure; Al = Sanilac hilum
area (50x); A2 = Sanilac micropyle (lOOOx); Bl
• Nep-2 hilum area (50x); B2 = Nep-2 micropyle
(lOOOX); Cl = San-Fernando hilum area; C2 =
San-Fernando micropyle area (lOOOx) . . . • • . .
132
30
Scanning electron micrographs of soaked bean
cotyledon structures (400x and 1000x) Al and A2
• Sanilac cotyledon; Bl and B2 = Nep-2 cotyle-
don; Cl and C2 = San-Fernando cotyledon • . . • .
133
31
Scanning electron micrographs of processed
Sanilac seed coat outer surface structure; A and
B = cuticle and intermembrane outer surface
(lOOX and 400x); C = cuticle cell outer surface
(3000x); D = protein intermembrane cells outer
surface (3000x); PC = protein intermembrane
cells; CU = cuticle • . . . . . . . • . . • . • .
135
32
Scanning electron micrographs of processed Nep-2
seed coat outer surface structures; A and B =
cuticle and intermembrane surface (lOOx and
400x); C = cuticle cell outer surface (3000x);
D = protein intermembrane cells outer surface
(3000x); PC = protein intermembrane cells; CU =
cuti cl e .
. .
.
.
.
. .
. .
.
.
. .
.
.
.
.
. .
.
136
33
Scanning electron micrographs of processed San-
Fernando seed coat outer surface structures; A
and B (lOOx and 400x) and C and D (3000x) =
cuticle cell outer surface; CU = cuticle • • . . •
137
34
Scanning electron micrographs of processed bean
structures; Al and A2 = Sanilac cotyledon (400x
and 2000x); Bl and B2 = Nep-2 cotyledon (400x
and 2000x); Cl and C2 = SF cotyledon (400x and
2000x); RS = expanded rupture starch granules • .
139
35
Mean Hunter color and color difference values of
dry and processed bean cultivars (Sanilac,
Nep-2 and San-Fernando) over three crop years
(19~~, 1979 and 1980) . . . . . . . • . . • .
142
xii i

---

Figure
Page
36
Percent water content for bean cultivars
(Sani1ac, Nep-2 and San-Fernando) in qua1ity
parameter eva1uation over three crop years
(1978,1979 and 1980) . . . . . . . . • • • .
144
37
Drained weight for bean cultivars (Sani1ac,
Nep-2 and San-Fernando) in canned qua1ity para-
meter eva1uation over three crop years (1978,
1979 an d 1980) . . . . . . • . ',' . . . . . . .
147
38
Mean Kramer Shear Press curves for bean culti-
vars (Sani1ac, Nep-2 and San-Fernando) over
three crop years (1978, 1979 and 1980) • . . . .
148
39
Texture for bean cultivars (Sani1ac, Nep-2 and
San-Fernando) in canned bean qua1ity parameter
eva1uation over three crop years (1978, 1979
149
and 1980)
.
Percent water absorption pattern of dry bean
cultivars (Sani1ac, Nep-2 and San-Fernando)
soaked in ambient temperature water (1:1, 'tap
water:disti11ed water) for up to 90 minutes for
crop year 1978 . . . . • . . . . . . . . • . . .
179
Percent water absorption pattern for dry bean
cultivars (Sani1ac, Nep-2 and San-Fernando)
soaked in ambient temperature water (1:1, tap
water:disti11ed water) for up to 90 minutes for
crop year 1979 . . . . . . . . . . . . • . . . .
180
'Percent water absorption pattern for dry bean
cultivars (Sani1ac, Nep-2 and San-Fernando)
soaked in ambient temperature water (1:1, tap
water:disti11ed water) for up to 90 minutes for
crop year 1980 . . . . . • . . . . . . . . . . .
181
B1 Past1ng curves of dry bean f10urs (who1e bean
and cotyledon) by cultivars (Sani1ac, Nep-2 and
San-Fernando) in a 400 ml phosphate buffer
solution at pH 5.30 for crop year 1978 . . • .
183
~
Pasting curves of dry bean f10urs (who1e bean
and cotyledon) by cultivars (Sani1ac, Nep-2 and
San-Fernando) in a 400 ml phosphate buffer
solution at pH 5.30 for crop year 1979 . . • • .
184
xiv

---

Figure
Page
Pasting curves of dry bean flours (whole bean and
cotyledon) by cultivars (Sanilac, Nep-2 and
San-Fernando) in a 400 ml phosphate buffer solu-
tion at pH 5.30 for crop year 1980
.
185
Cl
Hunter color and color difference values of dry
and processed bean cultivars (Sanilac, Nep-2 and
San-Fernando) for crop year
187
1978
.
Hunter color and color difference values of dry
and processed bean cultivars (Sanilac, Nep-2 and
San-Fernando) for cr op year
188
1979 . • . . . . . . .
Hunter color and color difference values of dry
and processed bean cultivars (Sanilac, Nep-2 and
San-Fernando) for crop year
189
1980
.
Dl
Percent water content of bean cultivars (Sanilac,
Nep-2 and San-Fernando) in quality parameter
evaluation for crop year 1978
.
191
Percent water content for bean cultivars (Sani-
lac, Nep-2 and San-Fernando) in dry, soaked and
canned beans quality parameters evaluation for
crop year 1979 . . . . . . . . . . • . . . . . . .
192
Percent water content for bean cultivars (Sani-
lac, Nep-2 and San-Fernando) in quality parameter
evaluation for crop year 1980 . . . • . . • . • •
193
E
. Drained weight for bean cultivars (Sanilac, Nep~2
and San-Fernando) in canned bean quality para-
meter evaluation by crop years (1978, 1979 and
194
1980)
.
,
.
.
.
.
.
.
.
.
.
.
•
.
.
.
.
.
..
.
.
•
Texture for bean cultivars (Sanilac, Nep-2 and
San-Fernando) in canned quality parameter
evaluation for crop year 1978
.
195
Texture for bean cultivars (Sanilac, Nep-2 and
San-Fernando) in canned quality parameter.
evaluation for crop year 1979 . . . • . . . . . .
196
Texture for bean cultivars (Sanilac, Nep-2 and·
San-Fernando) in canned bean quality parameter
evaluation for crop year 1980 . . . . . • . . • .
197
xv

---

INTRODUCTION
The World Food Problem
The major problems facing the world today are two-fold:
one of population; the other of not enough food.
Modern
medic;ne and improved sanitation have brought about a
spectacular increase ;n the average life span while the
birth rate in many countries remains practically unchanged.
The result is the much-discussed population explosion. 'In
developed countries, it is recognized that the difference
between birth rate and death rate is small due to birth
control and high medical practices while in developing and
lesser-developed countries that difference is larger due to
the fact that birth rate is highly by lack of birth control.
and also the death rate reduced by improvement of sanitation
and better medication availability.
Efforts to check the birth rate and to increase the food
supply are severely handicapped by lack of education.
adherence to customs. and lethargy from malnutrition and
undernutrition.
The problem of population and food is not new; it has
confronted man for nearly all the million years that human
beings have been on the earth.
The present problems differ
1

---

2
only in magnitude and intensity from problems that have been
faced many times before.
TodaYt one-half billion people out
.
of four billion (12.5% of the world population) do not
receive close to an adequate quantity of food; moreover t
possibly one-half the world is marginally fed.
This
non-availability of food to that portion of the world
population, mainly Latin America, Africa and Asia, could be
associated to many factors such as drought t infertility of
the so11s, inadequate farming lands (Sahel, Africa subsa-
har1an regions), inadequate food production due to the fact
that the farmer produces food only to feed his family and
not for commercial purposes; lack of mechanical means for
large production; lack of preservation, transformation and
transportation means of the food; political and economical
limitations; low salary leading to inability to buy enough
food and food of adequate aesthetic quality; and lack of
nutritional information and improvement of quality and yield
of the food available.
All those factors.contribute to the
low calorie intake of the one-half billion of the world
population.
Coupled with the general food shortage is the shortage
of specific nutrients.
There are six main categories of
nutrients that are needed to supply energy, promote growth
and repair body tissues and to regulate body processes,
These are water, carbohydrates t protein, lipid, vitamins
and minerals.
Those nutrients are quite present in a wide

---

3
variety of food but may be limited in sorne others.
8y lack
of nutritional information and inadequate distribution, one
may be exposed to inadequacy of one of those nutrients
leading to different sickness.
One such nutrient that has
received considerable attention is proteine
IIProtein Problem ll
The deficiency in animal protein in the diet deficit
regions of the world has been generally associated to poor
physical development of adults and even more markedly to
the protein deficiency diseases of children.
Kwashiorkor
and marasmus account for a very high child mortality between
weaning and 6 years of age and often leave the children that
survive with stunted physiques and retarded mentality.
Differentauthorities have set different goals for
animal protein in the diet.
The U.S. Department of Agricul-
ture would provide 10 grams of animal protein per person per
day, and 10 grams of pulse protein, the latter from
increased cultivation and use of leguminous crops and from
increased human consumption of the press cake from oil
seeds.
Cook (1962) set a goal of 16 grams of animal protein
per person per day for the under-nourished countries.
Pawley (1963), looking to the year 2000, set a goal of 20
grams of animal protein per person per day with the rest
of the requirement from other food sources.

---

4
In view of the paramount importance of a complete pro-
tein in the diet, particularly for child development, it is
not quite easy to accomplish the goals.
Four causes of
this t1 pro tein problem tl have been reported by the Protein-
Calorie Advisory Group (PAG) of the United Nations (PAG
Statement No. 20, 1973) as:
1)
Low wages and income and under-employment or un-
employment in rural or urban centers, all of which
limit the pure hase of the relatively costly foods
that contain protein of good quality.
2)
Difficulties associated with the production of
protein-rich food of animal or plant origin because
of ecological and agricultural limitations with
the result that they are usually expensive and
relatively short supply.
3)
The lack of effective food processing, distribution
and marketing system resulting in large loss of
food crops.
4)
The lack of knowledge of food values and food
preparation for children and specifie prejudices
against giving sorne protein foods to young children,
especially when they have infectious disease.
Possible Solutions to the t1protein Problem tl
Several approaches have been proposed to solve the pro-
tein gap but one should recognize that the situation is
very complex.
Altschul (1969) mentioned that the
conventional approach to increasing food supply is
to increase grain production,.and that to increasin~
protein quality of a diet is to increase animal produc-
tion.
He proposed two ways beyond conventional means:

---

5
1) Raise the protein impact of the major contributor to
protein in the diet; 2) develop new nutritious foods to
supplement those now available from conventional sources.
His views are directed toward synthetic amine acids,
vegetable protein mixtures, irnproved cereal products,
protein beverages, texture foods and others.
He suggested
that the major increase by far in protein supply for the
near future will come by increase in total supply of cereal
grains and the next will come by increase in conventional
protein sources - legumes and animal proteine
The United Nations Advisory Committee on the Application
of Science and Technology to development (1968) suggested
seven policy directions.
1.
Promotion of increased quantity and quality of
conventional plant and animal protein sources
suitable for direct human consumption.
2.
Improvement in the efficiency and scope of both
marine and fresh-water fisheries operations.
3.
Prevention of unnecessary losses of proteinaceous
foods in field storage, transport and home.
4.
Increase in the direct food use of oilseeds and
oi1seed-protein concentrates by human population.
5.
Promotion of production and use of fish-protein
concentrate.
6.
Increase in the production and use of synthetic
amine acids to improve the quality of protein in
cereals and other vegetable sources, and the develop~
ment of the use of other synthetic nutrients.
7.
Promotion of the development of single-cell protein
for both animal feeding and direct utilization by
man.

---

6
Kahn (1981) reported that the world food needs
for 1985 are estimated to be 44% greater than that
required in 1970, and about 70% of that 1985 estimate will
be needed to feed the Third World~
His approach to solve
the problem is a multisectoral coordination.
First, slowing
of population growth should be a paramount objective of any
developing country's program to alleviate hunger.
Second,
the choice of strategy that should be undertaken for
eradicating poverty associated with low protein intake
should be left to each government.
But it must be recog-
nized by planners, economists and policy makers that
increasing national income alone will not solve the poverty-
hunger problem.
Last, he stated that food production yield
per acre could be improved in many areas of the world, even
though factors such as climate, water, soil, energy, etc.,
influence productivity.
Of all the suggestions that have been made for solving
the protein problem, the one that has received the most
attention is the improved production and utilization of
conventional animal and plant protein sources.
Animal
products are ideally suited for meeting the protein needs
of people through out the world but they are costly, scarce
or not utilized fully because of ignorance, taboos or
poverty.

---

7
Needs of Vegetable Protein
Vegetable protein which are the basic plant protein
sources, typically supply over 80% of dietary protein in
developing countries.
Cereals and legumes give the largest
contributions, and they are utilized in various forms in
different regions.
Protein content among cereals ranges
from 6 to 14%; rice is low and wheat is high among the
predominant cereals eaten by humans.
Within each variety
there is a range in protein content; in wheat alone the
range can be 8 to 14%.
The quality of the cereal protein
is not as good as that from animals, primarily because the
amine acid pattern is deficient in one or more amine acids,
principally lysine; others are tryptophan, threonine and
methionine.
Because of that, attention has been paid to legume
proteine
Legumes grown in different parts of the world
enjoy widespread us~, have a high degree of acceptability,
relatively low cost and possess good nutritional proper-
ties.
They are known to be rich sources of lysine but
deficient in sulfur amine acids.
This places them as
natural complements to cereal based diets.
In general, the aim to the solution of the protein
problem is the production of protein rich foods with1n the
developing countries or regions.
Several indigenous crops
which are still unknown to the western world need to be

---

8
investigated in order to eva1uate their nutritiona1 status
and incorporate that into local agricu1tura1 systems.
Grain
1egumes which fa11
in that category have become important
crops.
Yie1d Prob1em
Carperter (1981) reported that wor1d production of grain
1egumes relative to that of cerea1 grains is fa11ing, main1y
because of considerab1y greater yie1ds obtained with cerea1.
Yet, governments and international agencies continue to
encourage production of the se 1egumes because they "fix"
nitrogen, and because of a be1ief that they make a special
contribution to the diet of people unab1e to afford high
1eve1s of mi1k and other animal products.
In the U.S., it has been shown that average bean yie1d
has a downward trend in the past 20 years suggesting that a
barrier to higher yie1ds has been reached in beans.
In
Michigan, through Michigan Improved Sean and Seed Program.
Michigan State University has been made responsib1e for the
deve1opment, eva1uation and re1ease of new varieties,
Hosfie1d and Uebersax (1980) stated that there
is a number
of reasons for yie1d barri ers in crops, but the 1east
common denominator is the variety because variety possesses
a certain genetic potentia1 or program prescribed by its
,genetic complements to produce under a given set of circum-
stances.
They exp1ained that when circumstances are

---

9
~.
optimum, the variety will produce at a maximum, and if
c1rcumstances change and prevailing conditions limit the
genetic potential of a variety, the variety then must be
replaced with one that can maximize its genetic potential
in terms of productivity.
This replacement of variety could
be important for the seed quality.
Frazier (1978) indicated
that sorne of the highest bean yields being achieved in
Michigan-wide trials are for beans with tropical germplasm
in their pedigrees.
Smucker et !l. (1978) precised that
preliminary results from several field experiments suggest
that black beans derived from tropical accessions have root
systems more tolerant to partial soil compaction and water
stress.
Bean Production and Utilization
Figure l shows the family and subfamily of beans.
The
papilionaceae sub-family grown in tropical and subtropical
regions provide the most edible beans.
The subject of this dissertation is Phaseolus vulgaris L.,
the co~mon dry bean which is the grain legume consumed in
greatest quantity in the world as a whole, and it exempli-
fies all the problems associated with these materials.
Because of the high nutritional value of beans, various
workers have conducted compositional analyses: investiga-
tions generally include protein, fat, moisture, fiber, ash,
mineral, vitamin, carbohydrate and texture.

---

Leguminaseae Family
+
i l
.
Sub-family
•
Minosaceae
Caesielpinaceae
Papilionaceae
\\I---------..v
1
Grow in tropical regions
Grow in tropical and
only.
Only a few provide
temperate regions.
human foods.
Provide human goods.
!
Tribe
+,..........-----.
..
Aeschynomeneae
Phaseoleae
. Vicieae
Groundnut
Pigeon pea
Chickpea
Soybean
......
Lima bean
a
Mung bean
Common bean
Cowpea
Figure 1.
Phylogenetic relationship of seyeral dry edible legumes.

---

11
Dry beans often require long cooking periods to become
eating soft.
Hydration of the beans prior to cooking is an
important factor in the legume industries.
Conventional
methods of preparing and cooking dry beans involve soaking
overnight in water at room temperature (16-24 hrs) followed
by cooking in boiling water for 45 min to 3 hrs.
Time of
cooking depends on bean variety and storage history.
Many
investigations have been carried out to solve that problem
and it has been suggested that information on chemical
composition of specific tissues and localization of chemical
constituents in those tissues is a prerequisite for an
explanation of physical and chemical changes in bean tissues
during mechanical. thermal. chemical and enzymatical treat-
ment (Powrie ~!l.. 1960).
Texture is an important quality
characteristic of cooked beans.
One should also bear in
mind that texture of adequate acceptability by consumer is
of quite importance in bean production and now to plant
breeders.
Although there have been numerous attempts to
measure objectively the texture of cooked beans. there is
very little information on the texture of raw and soaked
beans.
Beans have also been associated with excessive intesti-
nal gas production known as flatus.
Studies have been
directed toward elucidating t~e cause of flatulence in
beans.
Several indigestible su gars namely stachyose and
raffinose have been implicated in the flatus associated

---

12
with eating cooked dry beans.
To study this problem,
research has been directed toward:
1.
characterizing the specific component.
2.
developing physical and chemical means of extrac-
ting these components.
3.
developing practical cooking or processing tech-
niques which may be used to reduce the problem.
4.
attempting to identify genetic variants possessing
low levels of oligosaccharides which may be used
to biologically remove the flatulence ,factors of
beans.
Although yield is the most important solution criteria
in bean breeding, the breeder is cognizant of consumer
preferences when introducing new cultivars.
Consumers have
acquired specific preferences for various combinations of
visual characteristics of seeds and, beans are generally
soaked and must be cooked to make them tender and palatable
and to render seed protein nutritionally available before
eating (Rockland and Jones, 1974).
The use of modern technology has provided the beans by
which food quality evaluations may be conducted with a high
degree of objectivity and precision on small amounts of seed
,
(Hulses ~~., 1977).
However, little information is
available as to what characteristics lead to the variability
for physical and chemical characters noted in dry edible
beans (Hosfield and Uebersax, 1980).

---

13
Objectives
Strains that differ from each other genetically at one
locus only are referred to as isolines.
Since IINep-2 11 was
the result of an induced mutation in IIS an -Fernando ll that
presumably only affected the color locus in common bean
(Moh, 1971), the differences in physico-chemical properties
were surprising and prompted this investigation because of
the possible impact on cooking quality.
Earlier processing
evaluations of these strains showed that IIS an -Fernando ll and
IINep-2 11 differed in the rate of water absorption during
soaking and IIS an -Fernando ll required almost 2 times the force
as IINep-2 11 to shear a sample of beans to the point of
deformation using a Kramer Shear Press Instrument,
The objective of this research was to investigate
physico-chemical differences noted between the above iso~
lines.
The dry bean cultivar, Sanilac, was ~sed as a point
of reference (standard) for all evaluations,
For this
purpose experiments were designed to ascertain the:
1.
Proximate composition of the two iso11nes and
Sanilac (a commercial standard Phaseolus vulgaris L.
bean).
2.
Possible flatulence sugars for potentiality of
reduction of the levels through breeding.
3.
Microstructure of the three cultivars of beans by
scanning electron microscope (SEM) and relate
differences to texture.
4.
Hydration rate of the three varieties and relate
that to their microstructures.

---

14
5.
Physical properties (viscosity) of the flour from
the beans and relate that to pasting characteris-
tics.
6.
Processing quality and texture of the three cultivars.
7.
Effect of soaking and cooking on the microstructure
of the beans and relate that to texture.

---

LITERATURE REVIEW
Nutritional Contribution of Food Legume Seeds
to World Hunger Problem
Legume seeds are grown and used for food in nearly all
the temperate and tropical areas of the world.
Rockland
and Nishi (1979) mentioned that 12 varieties have commercial
importance and legumes require less energy per unit protein
production than cereal grains and particularly animal pro-
tein.
Tobin and Carpenter (1978) in their critical review
of the literature concerning protein quality evaluations and
other properties of the common beans, Phaseolus vulgaris L.,
reported PER (Protein Efficiency Ratio) values for varieties
and cultivars ranged from 1 •.0 to 2.0.
Rockland and Nishi
(1979) reported values of 1.2 - 1.4 for authentic strains of
cooked standard and 1.2 - 1.6 for analogous quick-cooking
products prepared from various f. vulgaris L. cultivars.
Rockland (1978) reported that a maximum PER for quick-cooking
beans was observed after cooking for 5 min in boiling water.
After cooking for 15-30 min, the PER values were slightly
lower.
Standard-cooked beans, cooked for 5 min, had about
the same PER as the quick-cooking beans.
There, the time
and energy factors are involved in the efficiency of the
l 5

---

16
bean protein.
Tobin and Carpenter (1978) suggested that
extended cooking, especially at higher pressure and tempera-
ture, tends to 10wer nutritional quality.
Eliasetal.
(1979) and Rockland and Radke (1981) obtained 10wer PER
values for beans and cook water combined than for drained
beans.
They suggested that seed-coat tannins in colored
beans may prevent complete utilization of legume proteins.
Beans, generally, are fairly good sources of riboflavin
and vitamin E and others such as thiamin, niacin. vitamin
B6 ,and folacin (Bunnel ~~.• 1965; Harris et ~•• 1950;
and Patwardhan, 1962).
Guerrant et~. (1946), Lamb ~~.
(1946), Lantz (l938) and Shroeder (l971) reported
riboflavin retention of 72 - 105% in processed bean.
Bunnel
~~. (1965) and Harris et ~. (1950) reported vitamin E
10sses of 70 - 90% in canned beans.
Normal bean processing
methods may not prevent the formation of hydroperoxides
which allow the tocopherol side-chain to be oxidized and
further degraded to peroxides which in turn further decom-
pose to cleavage to aldehydes and ketones.
Riboflavin was
higher in steam blanched beans but vitamin E was not
affected since vitamin E is not water soluble (Connie ~ !l.,
1979) using navy beans, Pinks, Pinto and snap beans.
As
storage time increased, the vitamin retenti on decreased.
Nordstrom and Sistrunk (1977) reported vitamin E 10ss in
canned bean during storage.
Bunnel ~~. (1965) and others
( Am es, 197 2; Ha r ris ~ al., 1950 ) r e p0rte d ri bof 1a vin los ses

---

17
of 90% in canned bean during storage.
Beans are also source of minerals such as iron, calcium,
phosphorus, magnesium, sodium, potassium, manganese and
copper.
Augustin ~~. (1981) reported retention of those
minerals ranging from 38.5% to 103.2% with sodium and
calcium being the lowest and highest, respectively.
All the above factors are of great interest to plant
breeders in the development of a new variety.
Cooking and'Processing
Legume seeds often require long cooking periods to
become soft.
Cooking is necessary not only to tenderize
the seed coat and cotyledon and develop acceptable flavor
and texture, but also to make the bean protein nutritionally
available.
Factors affecting cooking characteristics have
been associated with seed coat (Synder, 1936; Gloyer, 1932)
and cotyledon (Mattson, 1946).
Adams (1975) by relating
soaking time and cookability mentioned that the hilum and
micropylar areas usually admit water readily, but seed coats
differ strikingly in this regard.
What he did not specify
was the structural characteristics of the micropyle and
seed coat of the beans he used.
He stated that both the
age of the seed and the genotype of the maternal parent are
important regulatory factors.
Powrie et al. (1960) stated
- . - . -
that information of chemical composition of specific tissues
and localization, of chemical constituents in those tissues

---

18
is a prerequisite for an exp1anation of physica1 and chemi-
cal changes in bean tissues during mechanica1, thermal,
chemica1 and enzymatic treatments.
The mechanica1 factors are main1y seed coat breakage and
sp1itting of cotyledons resu1ting from hand1ing in harvesting
and c1eaning.
Because of temperature, rainfa11 and other
weather conditions during growth, dry beans are subject to
cracks, hard seed coats and other prob1ems that can affect
processing procedures (Connie ~~., 1979).
Sp1its were
affected by bean type, initial moisture content and storage
time.
Adams and Bedford (1975) stated that, as a genera1
ru1e, the 1arger and more irregu1ar shaped seeds are the
more sensitive to mechanica1 abuse than are the sma11er,
more near1y rounded seeds.
Bressani and Elias (1974) reported a minimum of two
hours for cooking soaked dry beans (Phaseo1us vu1garis L.)
at atmospheric pressure.
There have been numerous attempts
(Esse1en and Davis, 1942; Feldberg ~ ~., 1956; Dorsey
et ~., 1961) to find a way of lowering the cooking time
for 1egumes.
An important deve10pment in the uti1ization
of who1e 1egumes has been the preparation of quick-cooking
1egume products.
Steinkraus et ~. (1964) reported on a new process for
preparation of quick-cooking dehydrated beans by hydrating
the dry beans through soaking in water for 15 min, fo11owed
by a precooking in steam and coating by dipping in a 20%

---

19
o
sucrose solution at 160 F, then dehydrating.
Rockland and
Metzler (1967) reported a process for quick-cooking large
dry lima beans using an intermittent vacuum treatment for
30 to 60 min in a solution of inorganic salts (sodium
chloride, tripolyphosphate, bicarbonate and carbonate),
soaking for 6 hours in the same salt solution, rinsing and
drying.
They indicated that their process facilitated
infusion of the salt solution through the hilum and
fissures in the hydrophobic outer layer of the seed coat.
Wetted by the solution, the inner membrane hydrates rapidly,
plasticizing the seed coat and causing itto expand to its
maximum dimensions within a few minutes.
As a result,
cotyledons imbibe the solution rapidly.
This causes about
80% reduction in the cooking time.
Another method of
processing legume seeds involves deh~lling of the beans
prior to cooking.
Practices of dehulling differ from legume
to legume and with the same legume in different parts of the
world.
Although the processing of legumes for home consump-
tion with or without dehulling differs to a considerable
extent in different countries and regions, the similarities
outweigh the differences (Aykroyd and Doughty, 1964).
Sorne
other methods are fermentation, roasting, parching, agglo-
meration and germination, but the iÏ!mportant one is the hard-
to-cook phenomena.

---

20
Moisture Factor
For processing, the generally desired moisture content
in dried beans is in the range of 14 - 18%.
USDA (1975)
permits moisture content up to 18% without being designated
as IIhigh moisture beans ll •
Morris and Wood (1956) reported
that beans with a moisture content above 13% deteriorate
significantly in texture and flavor after 6 months at 25 0 C.
Beans stored at less th an 10% moisture maintained good
quality and stored well even after 24 months.
Too low
moisture content can also crea te other processing problems
such as not imbibing water normally; bean seed coats becoming
brittle, thus being subjected to cracking.
But Connie et al.
(1979) reported that low original moisture level before
soaking resulted in higher hydration ratios in all beans
used (Pinks, Red kidney, common navy, hite, Pinto, snap
beans) except Pinks and Avenger.
Bean samples containing
16% initial moisture were firmer in texture after thermal
processing.
Burr and Morris (1968), Kon (1968), Morris and
Wood (1956), Muneta (1964) and Ruiloba (1973) reported that
the use of a low storage temperature (4 0 C) or the practice
of storing beans with a low moisture content (8-10%) at a
relatively low humidity environment has been shown to
minimize the development of a hard-shell condition in legume
seeds, including black beans.
Adams and Bedford (1975)
stated that when various types of beans at about 12.5%
moisture level are dropped 10 meters ante a slanting steel

---

21
surface, they usually will incur sufficient damage to permit
detection of genetic differences.
Morris (1964) studied the
effect of moisture content and temperature during storage on
the cooking time of Pinto, Sanilac and lima beans.
He found
that cooking time increased with storage time, especially at
moisture contents above 10%.
At 13% moisture content the
cooking time after 12 months was three times the initial
cooking time.
At 10% the cooking time after 12 months was
only slightly longer than the initial time.
Cooking time
also increased with storage temperature, especially at high
moisture contents.
Barriga-Solorio (1961) reported that at
9.7% moisture 27.8% of Sanilac seed were damaged but at
15.5% moisture damage was reduced to only 5.3%.
Muneta
(1964) found a correlation of +0.80 between moisture content
of stored dry beans of four cultivars and cooking time.
He
thought moisture content per se was not the primary factor
in determining cooking time and postulated that factor or
factors connected with high moisture content resulted in a
longer time to breakdown (TTB) phenomena.
From all the
above views, the speculation of Muneta (1964) gives space
for identification of other factors either chemically or
structurally in the understanding of the hard to cook beans
phenomenon.
Hydration
It is well known that when a sample of beans is soaked
in water at room temperature, sorne of the beans do not

---

22
imbibe water, presumably because the seed coats are imper-
meable (Burr, 1973).
This condition is referred to as
hard-shell or seed-hardness.
Hydration of the beans prior to cooking is an important
factor in the legume industries.
Conventional method of
preparing and cooking dry beans involves soaking overnight
in water at room temperature (16-24 hours) followed by
cooking in boiling water for 45 min to 3 hrs (Oguntunde,
1981).
Time of cooking depends on bean variety
and storage history.
Inadequate soaking lengthens cooking
time and affects the cooked bean texture.
Longer cooking
times lead to the use of more energy th an necessary in the
processing of beans.
Adams (1975) mentioned that hydrating seed to 53 to 57%
will insure uniform expansion in the can during the thermal
process and insure product tenderness.
He specified that a
long, soft-water soak will leave the bean more tender or
even mushy.
Hard water tends to toughen the skins and firm
the texture of the cotyledon.
He suggested that water
containing 25 - 50 ppm calcium is considered optimum.
He
added that if too-hard water cannot be softened chemically
or by ionic exchange resins, the firm effect can be counter-
acted by addition of O. l to 0.2% of sodium polyphosphate to
the soak.
Gloyer (1932), Morris ~~. (1950), and Steinkraus
~~. (1964) reported a favorable effect of a heat
\\

---

23
treatment on the water absorption of beans which minimized
the hard-she11 deve10pment in the grain.
However. other
authors (Burr and Morris, 1968) reported that beans that
rehydrate as quick1y as normal beans usua11y need a pro-
longed cooking time, thus indicating no correlation between
water absorption capacity and cooking time.
These findings
revea1 that, at 1east in sorne bean varieties, a higher
water absorption capacity (lower hard she11) is not neces-
sari1y corre1ated with a shorter cooking time.
Molina et al.
(1976) indicated that heat treatment did not affect the
physica1 appearance of black bean grains (Phaseo1us vu1garis
L.) but significant1y (P$0.05) decreased the deve10pment of
the hard-to-cook phenomenon in foods.
Powrie et ~. (1960)
reported that the seed coats from soaked navy beans pos-
sessed an average moisture 1eve1 of 76.6%.
This high
capacity of the seed coat for water uptake suggested the
possibi1ity of water migration through the seed coat for
the hydration of other bean tissues during the soaking
periode
Snyder (1936) reported that for sorne bean seeds,
the entrance of water at ordinary temperature was 1arge1y
through the micropy1e and germinal area.
Ham1y (1932) sug-
gested that for sorne seeds. the micropy1e is probab1y no
more than a ho1e for ex chance of gases.
The importance of the hi1um in relation to the ripening
of the seed and the permeabi1ity of the testa in sorne
Papi1ionaceae was reported by Hyde (1954).
He found that

---

24
the hilum in sorne seeds performs a function essential for
the hardshell condition.
But it is not still very clear
what anatomical structures in the legume seed are respon-
sible for water absorption or the development of the hard-
shell condition.
According to Powrie ~.!l. (1960) most of
the water, both bound and free, presumably resides
in the
proteinaceous and cellulosic portions of.the cells.
In
general, one should recognize that the seed coat, hilum and
micropyle together may form an integrated specialized water
absorption/removal system.
To understand the slow hydration rate and the hard-to-
cook phenomenon, various investigations have been carried
out.
Most of the research and reports have been concentra-
ted on the relation of the external structures like hilum,
seed coat and micropyle to those characteristics.
Little
is done on the proteinaceous matrix and the cellulosic
portions of the cells.
Powrie ~.!l. (1960) raised the
question of a possible relation between these structures
and seed hardness.
There is no report on the effect of
proteinaceous matrix on the hardness of legume seeds but work
done on wheat indicates that there may be such an effect.
Scanning electron microscopes has commonly been used in such
studies.
In general, the primary determinant of wheat hard-
ness was shown to be genetically controlled and appeared to
relate to factors influencing the degree of compactness of
endos perm ce 11 components (Mos s ~ .!l., 1973; Greenaway,

---

25
1969; Stenvert and Kingswood, 1977).
Much more work needs to be do ne on the structural dif-
ferences of different varieties of legume seeds in order to
be able to show different water absorption and hardness
properties.
Texture Measurement Means
Texture measuring instruments can be divided into two
classes (Voisey, 1971a).
a)
Special purpose to perform a particular type of test
such as tension, compression, shear etc. on either several
products, or a specific type of product.
b)
General purpose to test a wide range of products
using a diverse range of methods.
To name a few:
- Brine flotation test - USDA method (Anon. 1945)
- Size of peas (Boggs ~ ~., 1942, 1943)
- Tenderometer (Mart"in, 1937)
- Texturometer (Lee, 1941 )
- Penetrometer (Anon. , 1938; Boggs et ~., 1942)
- Specific gravit y and density (Lee, 1941)
- Alcohol-insoluble solids (Kertesz, 1934)
- Total solids (dry matter) (Strasburger, 1933)
- Starch (Nielson et al., 1947)
- Sugars (Lee, 1941)
- Refractive index (Walls, 1936)

---

26
Most of those methods used were not reliable for measur-
ing tenderness and maturity of seeds (Makower, 1950).
He
considered the tenderometer to be probably the best means
of determining the maturity of peas.
For research, the general purpose type i s the logical
choice because of operational flexibility.
- Kramer Shear Press (Kramer et ~., 1951)
- Instron Universal Testing Machine (Bourne et ~.,
1966)
- General Food Texturometer (Friedman ~ tl., 1963)
- Ottawa Texture Measuring System (Voisey and deMan,
1976)
These have basic components common to all such devices:
a)
a mechanism for deforming the sample
b)
a system for recording force, deformation and time
c)
a test cell to hold the sample (Voisey, 1971b).
The test cell should subject the sample to appropriate
forces to determine the textural characteristics of interest.
All these instruments measure texture based on the response
of multi-bean sample.
The shear press was first used by Kramer and Asmlid
(1953) to test peas.
Other scientists have used the same
instrument to measure texture of vegetables (Ang ~ tl.,
1963) and beans (Rockland, 1964; Sanchez and Woskow, 1964;
Hoff and Nelson, 1964; Rockland, 1966; Luh et tl., 1975;
and Quast and de Silva, 1977a, 1977b).
According to Voisey

---

27
and Nonnecke (1973) the Kramer Shear Press is not entirely
suitable for use in a production environment because safety
features and fully automatic operation are not incorporated.
The advantages associated with the instrument is that the
force gauge, cell and shearing blades come as separate
attachments which can be easily transported for calibra-
tion and it is suitable for testing a wide range of foods.
The Instron Universal Testing Machine measures compres-
sive and tensile properties of materials and it has become
one of the most widely used texture measuring instrument
because the working parts of most texture measuring devices
used in the food industry can be used in that machine
(Sefa-Dedeh, 1979).
Several scientists have used also the
instrument on different types of food products (Bourne
~~., 1966; LaBelle ~ ~., 1969; Lee, 1970; Voisey and
Larmond, 1971).
The Ottawa Texture Measuring System (O.T.M.S.) was
• designed for testing foods to combine the advantages of
Kramer Shear Press and Instron machines and to eliminate
sorne disadvantages (Voisey and deMan, 1976).
A wide range
of test cells and attachments can be used in this machine
depending on the food being tested.
Other investigators have measured texture based on the
response of single beans (Malcom et ~., 1956; Ismael,
1976; Steinkraus et ~., 1964; Burr and Morris, 1968;
Molina ~~., 1976; Bourne, 1972; Ige, 1977).
The methods

---

28
used inc1ude a penetrometer, a1coho1-inso1ub1e solids, brine
f1otation and starch measurement.
Recent1y, Hudson (1982),
at the USDA Regional Plant Introduction Station at Washington
State University, has deve10ped a new method for measuring
cooking based on the response of single beans.
His apparatus
consists of 100 test points in a la x la array.
Each point
has a cup holding a single bean seed.
The method is as
described in the Materia1s and Methods of this dissertation
and it is based on time to breakdown (TTB) system with each
individua1 best looking bean seeds.
Sensory ana1ysis and objective measurement are a1so
used to conduct research into textura1 characteristics and
for production qua1ity control but using taste panels is
cumbersome, time consuming, expensive and requires suffi-
cient quantity of samp1e.
Re1ationship of Texture with Hard-to-cook Beans
Texture expresses three parameters (Adams, 1975):
a)
Firmness: measured by the force required to pene-
trate a substance.
Perceived on first bite.
b)
Gumminess: measured by the force required to
disintegrate a substance.
Perceived during chewing.
c)
Adhesiveness: measured by force required to remove
the materia1 from the mouth.
Perceived during
chewing.

---

29
These parameters could well be evaluated with reasonable
accuracy by a sensory panel and also measured quantitatively
by quite a variety of methods as discussed above.
Those
methods have been used by various investigators ta study
sorne basic quality characteristics of beans.
Powrie ~~. (1960) raised the question of possible
relation between the seed structure and hardness.
Rockland
and Jones (1974) suggested that the separation of bean cells
during cooking may be related ta the transportation or
removal of divalent cations, particularly calcium and
magnesium, from bridge positions within the pectinaceous
matrix of the middle lamella.
They also showed that there
is no breakdown of the cell wall of cooked bean.
Rather
they found that cooked or partially cooked bean cells
separated readily along the surface of individual intact
cell walls.
Short heat treatment loosened the intercel-
lular matrix of the middle lamella sufficiently ta allow
separation of individual cells without rupture of cell
walls.
Other scientists (Linehan ~ ~., 1969; Huges ~ ~.,
1975) attributed the hard-ta-cook phenomena ta the solubili-
zation and diffusion of starch from cells while plant
tissues are being cooked.
According ta Hahn ~~.
(1977),
starch granules in lima beans maintained slight birefringence
after cooking.
Varriano-Martson and de Omona (1979) indi-
cated that many of the starch granules of black beans

---

30
(Phaseolus vulgaris L.) exhibited sorne birefringence ev en
after long cooking periods.
Voisey (1971) working on the
measurement of baked bean texture found that large differen-
ces in texture are evident from year to year and between
recipe used.
It was also suggested that development of hard
shells can be the product of a chemical or enzymatic process
in the seed (Burr ~~., 1968; Kon, 1968; Morris and Wood,
1956; Muneta, 1964; Ruiloba, 1973).
Mattson (1946) reported that the cookability of different
dry pea varieties is related to their contents of phytic
acid content and calcium.
He suggested that when the phytic
acid content is low, the pectin in the middle lamella formed
insoluble calcium and magnesium pectates, causing the poor
cooking quality.
In general there is no clear understanding at the present
time of the mechanîsm that governs the water uptake in beans
and the factors that affect the tenderness or softness of
the canned product.
From that point, knowledge of the chemical composition
and characteristics of sorne major constituents in beans, and
also of the physical and structural properties of the seeds
and flours is necessary to help in the understanding of the
hard-to-cook phenomena.
That could also be effectively used
in breeding to improve yield of food and feed associated
with quality of the grain.

---

31
Scanning Electron Microscopy (SEM)
The concept of a SEM is credited to Knoll
(1935) who
suggested that characteristics of a sample surface could be
observed by focusing a scanning electron beam on the surface
and recording the emitted current as a function of beam
position.
Unlike the TEM, which uses ultra-thin sections,
the SEM samples would not be sectioned at all (Rasmussen and
Hooper, 1974).
The first functional SEM was constructed by
von Ardenne (1938) based on Knoll 's concept but the first
commercial instrument became available in 1965.
Today's
models provide images with resolution limits of 6 - 10 nm.
Buono (1982) stated the principle of SEM as:
1.
A concentrated beam of electrons is focused on the
surface of a_sample (Figure 2A).
Electron beam size
vary from 15 to 10,000 ~ and energy from l to 40 Kv.
2.
The electron beam penetrates the sample to depths of
0.2 to 2 um, depending on the energy of the incident
electrons and the atomic number (Z) or density of
the sample.
The incident beam electrons are called
the primary electron beam.
Multiple scatterings of
the primary electrons by the beam interaction with
the atoms of the sample surface causes the beam to
widen in diameter as it penetrates into the sample
(Figure 2B).
The width of this scattering is on
the order of 0.2 to l um.
The combination of lateral
beam spreading with depth produces an interaction
volume within the sample.
3.
The interaction of the primary electrons with the
atoms of the sample produces a variety of signals
including secondary electrons, backscattering
electrons, Auger electrons, X-rays and light (Figure
2C) •
4.
The signals generated within the interaction volume
have different escape depths which can be used to
tailor the analysis to specific regions of the

---

32
• 2A ,CO!,!CENTRATED BEAM
28
ELECTRON BEAM PENETRATING
'f;
,t~~.t. OF ELECTRONS -
, THE SAMPlE
li'
. ,,1
,..~"'l'.
~ ';
,2C
SIGNALS
PRODUCED
2D
SIGNALS GENERATED
:~t,i Secondary Electron.
f
AuOIf Electron
Characteri.t~c I-ray.
, f r . Continuum lI-ray.
\\.t;~
,,;
~ i.' ': Photo"'
,~lX !:' ,,1~:
'''-;~:'{
'f
~., t-
,
..'
...
~
"
Escape
Deplh
l "
Electran Ileam
Induced curr_nt
Specimen CIIrrent
Tran.mltted
Electrons
Figure 2.
The Princip1e of Scanning Electron Microscopy.
Source: Buono (1982)

---

33
sample by judiciously choosing the output signal
and by varying the energy of the primary beam
(Figure 2D).
The escape depths for secondary
electron is l to la nm, the depth of backscattered
electron is 0.1 to l vm and the escape depth for
X-rays is 0.5 to 2 ~m.
Based on those principles, the SEM is rapidly becoming
necessary to plant studies along with light microscopy (LM)
and Transmission Electron Microscopy (TEM).
A basic differ-
ence between SEM and other microscopes is the use of scanning
coils to drive the beam in Y direction while being deflected
in the X direction and the detection and display of low
energy secondary electrons (Rasmussen and Hooper, 1974).
SEM technology is progressing toward greater system
automation to the point of using stepper motor stages that
provides images and analyses of selected areas on a large
sample without the presence of the operator (Buono, 1982).
The essential features of the SEM are shown in Figure 3.
The instrument contains: (1) an electron gun which provides
an electron beam capable of being accelerated from l to 40
Kv; (2) a system of magnetic lenses to provide a means of
focusing a tiny spot of electrons from the source on the
specimen; (3) a specimen stage for holding the sample; (4) a
scan generator to provide a means of scanning the spot of
electrons across the specimen; (5) an electron collector
coupled with a photomultiplier and amplifier to provide.a
.means of detecting the response from the specimen; and (6)
a cathode ray tube (CRT) display system capable of being

---

34
electron
t---~ high voltage
source
Anode
::L
-Aperture
Scan
Generator
Condenser 2
Visual
Deflection
Coils
-
perture
~ Condenser3
1 CRT
Record
1~1Z(d Aperture
Grid
Specimen
Video
Collector
1o--oI-~
mp
250V
Light
Scintillator Guide
Photomultiplier
and Head Amplifier
12.5 kV
Figure 3.
Essential Features of SEM.
Source: Mee, cited by Kassel and Shih, 1976

---

35
scanned in register with the incident scan.
SEMs are practical tools that have moved out of the
laboratory into the production and quality control areas,
They have become easier to opera te and more cost effective
in providing information about surface and subsurface
properties of microelectronic devises.
In early 1970, Hearle (1972) provided a comparison
information between optical, direct electron and Scanning
Electron Microscopy (Table 1).
The advantages of SEM given
are:
a)
great depth of focus
b)
the possibility of direct observation of the
external form of real objects, such as complex
fracture surfaces, at high magnification thus
avoiding the necessity to make thin replicas for use
in direct transmission electron rnicroscopy
t)
the ability to switch over a wide range of magni-
fication, so as to zoom down to fine detail on sorne
part identified in position on the whole object
d)
the ease of operation, and the large space available
for dynamic experiments on the specimen
e)
the use, besides the secondary electrons, of scat-
tered primary electrons, light emission, X-ray
emission, current in th~ emission, and many other
responses to generate an image and obtain useful
information about the specimen.

---

Table 1.
Comparison information between optical, direct electron and scanning electron
microscopy (SEM).
Optical
Scanning electron
Direct electron
Resolution - easy
5 llm
0.211m
100 ~ (la nm)*
- skilled
0.2 )Jm
100 A (la nm)
la A (1 nm)*
- special
O. l iJm
5 A (0.5 nm)
2 A (0.2 nm)*
Depth of focus
poor
high*
moderate
Mode - transmission
yes
yes
yes
- reflection
yes
yes
not satisfact0rY
- diffraction
yes
yes
yes
- other
sorne
many*
no
Specimen - preparation usually easy
easy*
skilled, liable to
w
0)
artefacts
- range and
versatile
versatile
only thin,
type
real or repl ica
real or repl ica
or replica
- maximum
thick*
medium
very thin
thickness
for trans-
mission
- environment versatile*
usually vacuum but
vacuum
can be modified
- available
sma 11
la rge*
small
space
Field of view
large enough
large enough
limited
Signal
only as image
available for
only as image
processing*
Cost
low*
high
high
*Advantages over others; disadvantages are underlined.
Source: Hearle, J.W.
1972.

---

37
f)
the information can be processed in various ways
to present images in different forms.
Sorne of the disadvantages are:
a)
htgh cost
b)
1ack of high reso1ution
c)
the vacuum environment of the specimen
d)
inabi1ity to show up interna1 detai1 visible in
optica1 microscopy
e)
1ack of co10r response
Because of the advantages enumerated above, scientists
have used SEM in many areas (bio10gy, medicine, horticul-
ture, food science, microbio10gy ..• etc.) to understand
basic princip1es involved in many materia1s and products.
Many cerea1 scientists abandoned 1ight microscopy techniques
because SEM samp1es cou1d be prepared easi1y.
In the food area, a1though microscopy has been used for
many years, research articles on food microstructure attemp-
ting to relate structural and functiona1 characteristics of
food have become common on1y within the past 10 - 15 years.
As mentioned by Hansen and F1ink (1976), the structure of
1ndivid~a1 food components, as we11 as of the finished
product, p1ays an important ro1e in determining appearance,
f1avor, rheo10gica1 properties and keeping qua1ities~
To
study the se interre1ationships, the microscope in its
various modes is being used to a110w visua1ization of the
different heterogeneities in the structure of food systems.

---

38
On the use of cereals and oilseeds for food, SEM has
been used extensively (Pomeranz and Sachs, 1972; Stanley
tl~., 1976; Sullins and Rooney, 1974; Holf, 1970).
Hall
and Sayre (1971a) used SEM to show the surface characteris-
tics of starch granules from tender white, pinto and lima
beans.
They also worked on root and tuber starches, cereal
starches, legumes starches and other foods and reported
much information on the sizes, shapes and surface details of
these starches(Hall and Sayre, 1969~ 1970a, 1970b).
Wolf
and Baker (1972) used to investigate the cotyledon interior
of water-soaked soybeans and revealed that protein bodies of
l - 10jLin diameter exhibited a covering spongy network.
McEwen tl!l. (1974) used SEM to examine the cotyledon and
seed coat of faba bean seed.
They revealed that there was
no discontinuity in the thick seed coat.
Cross section of
the seed coat showed characteristic palisade, parenchyma,
tracheid, and hour-glass cells, similar to those of other
legumes.
Hahn et ~. (1977) used SEM to characterize
intracellular configurational changes of starch granules
during gelatinization of standard and quick-cooking lima
bean cotyledon.
Rockland and Jones (1974) studied the
effect of cooking on water-soaked and salt water-soaked
beans.
Other studies on foods using SEM include those of
Vix et ~. (1971) on cottonseed, Van Hofsten (1972), Gill
and Tung (1976), Stanley tl~. (1976) on rapeseed.
On food
starches, reports were published from Hood ~~.
(1974),

---

39
Shetty ~~. (1974), da Silva and Luh (1978), Dronzek ~!l.
(1972), Robutti ~~. (1973), Hill and Dronzek (1973),
Crozet (1977), Agbo et !l. (1979), Saio ~!l. (1977),
Bernardin and Kasarda (1973), Orth ~~. (1973), Watson and
Dikeman (1977), Badi et !l. (1976), and Chabot ~!l.
(1976).
The food items involved were cereals, roots or
tubers, legumes and banana.
Other food systems such as
meat and related products and dairy products have also been
studied with SEM (Schaller and Powrie, 1971, 1972; Stanley
and Geissinger, 1972; Jones et ~., 1977; Buma and Henstra,
1971; Eino, 1974; Kalab and Emmons, 1974; .Stanley and
Emmons, 1977).
The above reports are few among the numerous reports on
the use of SEM in food research.
That technique which is
progressing toward greater system automation is presently
used extensively
and it will continue to be so
because of the advantages it has over other microscopy
techniques.
But in general, the key to obtaining useful
information from the microscopes is the scientist's ability
to recognize which microscope, if any, to use (Varriano-
Marston, 1981).
Legume Seed Anatomy
Corner (1951) in his classification of leguminous seeds
specif1ed that legume seeds are generally of medium or
large size, more or less compressed and exalbuminous, with

---

40
large embryo and a hard, dry, generally smooth testa
(Figure 4).
He indicated that two microscopic characteris-
tics distinguished the leguminous testa: 1) the external
palisade, developed from the outer epidermis of the outer
integument, and 2) the hour-glass cells, developed from the
outer hypodermis of the outer integument and, in sorne cases,
from its ;nner epidermis.
He stated that any microscopic
particle with these features is apparently identifiable as
leguminous.
The different 'features of the testa found in various
legume genera and species are: cuticle, palisade, hypodermal
hour-glass cells, mesophyll, inner hypodermal hour-glass
cells, vascular bundles and subhilar tissue (Corner, 1951;
Chowdhury and Buth, 1970).
The'position and characteristics
of the se features are reported in Table 2.
The seed coat
surrounds two cotyledons which are the most important compo-
nent of the bean seed.
They are composed of parenchyma
cells containing starch granules embedded in protein matrix
(Bagley et ~., 1963; Harris ~ ~., 1975; Opik, 1966, 1968;
Powrie ~~., 1960; Yatsu, 1965) and vascular bundles (Opik,
1966) and protein bodies forming the protein matrix.
Many
scientists report that textural characteristics and nutri-
tive value of processed bean presumably are influenced to a
large extent by the size and shape of cells, dimensions of
the cell walls and the localization of chemical constituents
in the cotyledon.
These factors and a few definitive

---

41
.-----EPICOTYL
~-----HYPOCOTYL
~.....,..:.;~a-- NICROPYLE
X"J--- RADICLE
HILUN-----~-:-:-_+
COTYLEDON
~~~~~~-----SEED COAT
Figure 4.
Visua1 observation of 1egume seed structure.

---

table 2.
Legume testa feature characteristics.
Feature
Position
Characteristic
Reference
•
Cuticle
First outer layer
-Thin and smooth
Corner (1951)
-Smooth and rough
Chowdhury and Buth
(1970)
Palisade Cells
Second outer layer
-Irregular shape and
Chowdhury and Buth
height
(1970)
Hour-glass Cells
Third outer layer
-Shape varies from
Chowdhury and Buth
next to palisade
species to species
(1970)
and within species
some legumes have
single hour-glass
~
N
but few have two
Mesophyll Cell s
The remainder of the
Chowdhury and Buth
seed tests
(1970)

---

43
answers have been obtained to relate the ultra-structure
of legume seeds to the textural characteristics of the
processed product.
Further research in this area is
warranted.
Starch and Viscosity
Starch is one of the most abundant naturally occurring
organic compounds.
It is found in almost all plant tis-
sues, where it functions as a source of reserve energy,
which can be utilized gradually through enzyme actions.
Starch accumulates to high concentration (20-70%) in the
roots, tubers, fruits and seeds of many plants.
Chemically,
starches from all sources are carbohydrates, polymers of
a-D-glucopyranose units linked primarily by 1,4- and
1,6-g1ucosidic bonds.
Each glucose unit contains one
primary and two secondary hydroxyl groups which are respon-
sible for the hydrophilic properties of the starch.
Starch
is made up of two types of polymer chains: amylose, the
linear fraction (1,4 bonds) and amylopectin, the branched
chain fraction (1,4 bonds and 1,6 bonds).
The ratio of
amylose and amylopectin in starches also provides sorne
indication of their origine
The exact structural rela~
tionship between the two components in the starch grain
is not known, but the molecules are linked together to
form starch granule by hydrogen bonds (Hahn, 1969; Wurzburg,
1968) .

---

44
Starch granules are insoluble in cold water.
In a
humid environment, starches will take up moisture, but the
swelling which occurs is reversible.
When starch granules
in water are heated past a critical temperature, in the
range 60 - lOoC (gelatinization temperature, characteristic
of a specific starch), the hydrogen bonds which hold the
granules together begin to weaken, allowing the granules to
swell.
Since the gelatinization begins at the hilum, the
first indication of swelling is the loss of birefringence.
Then, as the amylose fraction is dissolved and leached out,
the granules begin to take in water and clarity and the vis-
cosity of the slurry begins to increase.
Eventually, the
granules, having become completely hydrated, may collapse
and break down.
The resulting starch " pas te" is composed
of granule fragments and molecules in solution.
The visco-
sity of the paste decreases as a result of this granule
breakdown.
On cooling the paste usually increases in
viscosity and decreases in clarity.
This results from
retrogradation of the linear (amylose) molecules, a process
whereby the amylose molecules tend to form rigid gels by
hy~rogen bonding.
The degree of retrogradation is dependent
on amylose content and the degree to which the amylose has
been solubilized during the heating cycle.
Cross-bonded
starches have their granule structures reinforced by
covalent bonds and the granule shows less tendency to swell
and retrograde (Wurzburg, 1968; Leach, 1965; Smith, 1964;
•

---

45
Collison, 1968; Mazurs ~~., 1957; Hoseney ~ ~., 1978).
Effective control and utilization of retrogradation is
necessary to obtain food products of good quality after
transport and storage.
Since each raw starch has its own
characteristic viscosity/temperature profile as a result
of its particular granular composition and structure, sorne
degree of control can be achieved simple by selection of
the correct starch (Sanderson, 1981).
The suitability of starch, natural or modified, for a
specific use depends on the functional properties.
The
functional properties would include, for example, ease of
cooking (temperature, and stirring energy to cook through
the swelling region), thickening power (final viscosity of
the paste after cooking and cooling, as a function of
concentration), and stability (resistance to thinning
resulting from stirring, pH or temperature change),
Because of the variety of changes which may occur in
starch paste during processing, many different methods have
been developed for following these changes in the paste and
estimating the functional properties of the starch from
them.
These methods fall into two categories, those invol-
ving direct microscopic observation su ch as monitoring
granule swelling, loss of birefringence or staining .
reactions (Collison, 1968; McMaster, 1964), and those
involving measurement of physical properties such as
swelling power, solubilization, sedimentation rate or

---

46
viscosity (Collison, 1968; Schoch, 1964; Smith, 1964),
The
most useful methods measure the viscosity of the paste
continuously during the standardized-cooking and cooling
cycle which stimulates a wide variety of processing condi-
tions (Smith, 1964),
The most common instrument used for
this purpose is the Brabender Amylograph,
This machine
continuously measures the viscosity of starch pastes and
flours while they are stirred and heated at a constant
rate, held at 95 0 C for 30 minutes and held at 30 0 C for 30
to 60 minutes (these methods have also been developed for
relating the viscosity data to the functional properties of
the starch) (Mazurs et ~., 1957),
These kinds of data are
available for most food starches such as cereal, roots,
tubers, banana and legumes (Mazurs et~"
1957; Kite
~~., 1957; Hahn,~~"
1977; Carson, 1972; Berry et ~.,
1979; Rodriguez-Sosa and Gonzales, 1975; Agbo et ~"
1979).
Legume starches which are of interest in this disser-
tation have been studied by several scientists for their
incorporation into bread, cookies and other product formu-
lations.
Hall and Sayre (1971a), Kawamura ~~, (1955),
Lineback and Ke (1975), McEwen ~~, (1974), Rockland and
Jones (1974) found good agreement among various legume
starch granules of the same species.
They reported that
legume starch granules were ellipsoid, kidney-shaped, or
irregularly swollen, with an elongated hilum and smooth

---

47
surface with no evidence of fissures.
The sizes were deter-
mined as diameter width and 1ength ranged from 12 to 36 ~m
and 12 to 40 ~m for navy bean and from 16 to 18 ~m and
16 to 48 ~m for pinto bean respective1y,
Brabender hot-paste viscosity pattern of various
starches appears to be determined not on1y by the extent of
swe11ing of the starch granules and the resistance of the
swo11en granules to dissolution by heat and fragmentation
by shear (Lineback and Ke, 1975), but a1so by the presence
of soluble starch, which is 1eached from the granule struc-
ture (Allen ~ ~., 1977; Miller et ~., 1973) and the
interaction or cohesiveness between the swo11en granules
(Leach, 1965).
Schoch and Maywa1d (1968) c1assified the
viscosity patterns of "thick-boil ing" starches into four
types.
Type A:
High-swe11ing starches, e.g., potato, tapioca,
the waxy cerea1s, and ionic starch derivatives.
The granules of these starches swe11 enor-
mous1y when cooked in water, and the interna1
bonding forces become tenuous and fragile
toward shear.
Hence the Brabender shows a
high pasting peak fo11owed by rapid and major
thinning during cooking.
Type B:
Moderate-swe11ing starches, e.g., normal
cerea1 starches.
Because the granules do not
swe11 excessive1y to become fragi1e~ the se

---

48
starches show a lower pasting peak and much
less thinning during cooking.
Type C:
Restricted-swelling starches, especially
chemically cross-bonded products.
Cross-
linkages within the granule markedly reduce
swelling and solubilization, and stabilize the
swollen granule against mechanical fragmenta-
tion.
Hence the Brabender curve shows no
pasting peak, but rather a very high viscosity
which remains constant or else increases during
cooking.
Type D:
Starches with highly restricted swelling,
especially "high-amylose" corn starches con-
taining 55 - 70% linear fraction.
Because of
the internal rigidity imparted by the high
content of associated linear molecules, the
granules of the starches do not swell suffi-
ciently to give a viscous paste when cooked in
water at normal concentrations.
Hence, the
amount of starch must be increased two- or
threefold to give a significant hot-paste
viscosity of Type C.
However, such high-
amylose starches give a Type A or B viscosity
pattern when cooked in media which cause
greater granule swelling, e.g., 0.1 N sodium
hydroxide.

---

49
The above two scientists found that navy bean, lentil,
yellow pea and garbanzo gave Type C Brabender curves.
Mung
bean starch showed a mixed viscosity pattern - Type C at
low concentration and Type B at high concentration.
Navikul
and D'Appolonia (1979) studying navy beans, pinto beans,
faba beans and mung beans reported that the amylogram
curves of the legume starches showed higher initial pasting
temperature and higher viscosity than did wheat starch,
which would indicate a higher resistance to swelling and
rupture.
No peak viscosity during the hold period at 95 0 C
occurred with any of the legume starches, indicating that
the paste was relatively stable and that the granules did
not rupture during stirring, which is not the case with
wheat starch.
Lineback and Ke (1975) reported that the
Brabender hot-paste viscosity patterns for chick pea starch
at concentrations of 5, 6, 7, 8 and 9% are similar to those
reported by Schoch and Maywald (1968).
They also indicated
that the Brabender hot-paste viscosity patterns for horse
bean starch are virtually identical to those obtained for
chick pea starch at the same concentration.
The gelatini-
zation temperature ranges were 63.5 - 65 - 690C for chick
o
pea and 61 - 63.5 - 70 C for horse bean starches.
Vose
(1977) working on smooth-seeded field peas reported that
pasting curves of smooth pea starch showed restricted-
swelling characteristics similar to those shown by chemi-
cally cross-linked starches.
Retrogradation of the cooked

---

50
pastes resulted in rigid, opaque, friable gel with a firmer
texture than corn gels.
The gels occurred in either acidic
or basic solutions, demonstrating characteristic differences
when compared with corn or wheat pastes.
Because of the importance of legume seed in food
supplementation and new product development, the pasting
characteristics and functional properties of several other
varieties and species of beans still need to be investi-
gated.
Indigestible Bean Sugars Problem
Although legume seeds have high nutritional value among
vegetable proteins, they contain a considerable amount of
oligosaccharides which have been implicated as factors
responsible for flatulence (Steggerda, 1968; Steggerda and
Oimmick, 1968; Hellendoorn, 1969).
Flatulence is a common
complaint even among healthy individuals and is one of the
most common causes of abdominal discomfort.
Flatus
generation has been associated with the absence of a-
galactosidase activity in the upper intestinal tract of
humans and animals and the fermentation process of the
oligosaccharides by microorganisms with production of gâs.
It has been attributed also to swallowed air, inhibition of
intestinal anhydrase, production of carbon dioxide from
pancreatic bicarbonate (Rockland, 1969).
Rackis ~!l.
(1970) reported that the flatus produced by fermentation of

---

51
dietary carbohydrate contained in the 10wer intestine may
cause nausea, dyspepsia, constipation, cramps, diarrhea and
discomfort.
He specified that the f1atus-forming factor
is main1y found in the 10w mo1ecu1ar weight carbohydrate
fractions of 1egumes which contain primary sucrase, raf-
finose and stachyose.
Takana et ~. (1973) and Cerning
~~. (1975) reported that 1egumes contain appreciab1e
amounts of a-ga1actosides of sucrase, particu1ar1y raffi-
nase, stachyose and verbascose.
Akpapunam and Markakis
(1979), Cerning-Beroard and Fi1iatre (1976, Naviku1 and
D' Appo10nia (1978), 01son ~~. (1975), Rackis (1975) found
appreciab1e amounts of those oligosaccharides in mature
seed such as beans - Ca1ifornia sma11 white, Great Northern,
navy, pinto, kidney, say, faba, lima, field, and mung; peas,
cowpeas; chick peas; pigeon peas; horse gram; 1enti1s; and
1upines.
Tanusi (1972) reported that mung bean, broad bean
and smooth pea have sma11 amounts of ajucose; reducing
sugar ranges from 0.06 ta 0.10% and nonreducing sugar from
5 ta 7.0% in broad bean (~. faba), mung bean (Phaseo1us
aureus) and kidney bean (t. vu1garis),
Naviku1 and
D'Appo10nia (1978) showed that the total sugar content is
higher in a11 1egume f10urs than in wheat f10ur.
The 1egume
f10urs contain high 1eve1s of sucrase, stachyose and
verbascose, but navy and pinto bean f10urs contain sma11
amounts of verbascose.
Reddy and Sa1unkhe (1980) presented
the chemica1 structure of those sugars containing

---

52
a-ga1actosido-g1ucose and a-ga1actosido-ga1actose bonds as
nonreducing sugars shown in Figure 5.
When those sugars
are ingested by humans, two enzymes (invertase and
a-ga1actosidase) are required for complete hydro1ysis.
Gitze1mann ·and Auricchio (1965) exp1ained that because the
human gastrointestina1 tract does not possess an a-gal ac-
tosidase enzyme and because mamma1ian invertase is an
a-g1ucosidase (Reddy and Sa1unkhe, 1980), the metabo1ic
fate of raffinose fami1y sugars is uncertain.
Schweizer ~!l. (1978), working with twe1ve mature
1eguminous seed crops, reported in their study using 70%
ethano1 extraction procedure and gas chromatography tech-
niques, a new compound pinito1 (3-0-methy1-D-chiro-inosito1)
and three isomers of the new disaccharide, a-D-ga1acto-
pyranosy1-pinito1, for which the name ga1actopinito1 was
proposed.
The chemica1 structures are presented in
Figure 6.
These sugars iso1ated from soya beans (Glycine
max) were a1so identified in chick peas (Cicer aristinum L.),
1enti1s (Lens escu1enta) and beans (Phaseo1us vu1garis L.).
Their amounts ranged from 0.2 to 0.9% and 0.03 to 0.8% of
dry weight respective1y.
Schweizer ~!l. (1978) stated
that being a-ga1actosides, the new disaccharides must be
taken into account when considering flatulence prob1ems.
Wursch (1977) proved the ga1actopinito1s being indigestib1.e
to human intestinal enzymes.
Schweizer et al. (1978)
specified that it is probable that gas chromatography peaks

---

~
I.prl
ÇHZ
OH
•
O~
OH
RAFFINOSE
i
-
1
01- D - GAL-(l_6)-OC-D-GLU-(I"'Z)-Il-D-FRU
.Dt-D-UY
~
j
~1l2
~CH2
HOCII,
CHZOH
H
OH
0
H
H
OH
OH
Il
OH
STACHYOSE
U1
W
1
1
()( -D - OAL- (1-6)- 0( -D -GAL-(I_6)_ or _D- GLU_(I_2)-#, - D_FRU
+~D-GAL/
C~
~H2~H2
/ '
1/2
~
CU~' ~
OH~ HOC~CH2011
f
~~OI'
~
H
VERBASCOSE
1
1
0( -D -
GAL- (1.... 6)- OC'- D -GAL- (1 ....,)- ()( -D-OAL- (1-6)- OC -D- OLU- (1-'2) - Il - D - F RU
Figure 5.
Structure re1ationships of the raffinose fami1y oligosaccharides.
Source:
Reddy, N.R. and Sa1unkhe, O.F.
1980.

---

54
-- )
GALACTOPINITOL
ISOMER
OF GALACTOPI NITOL
0( -
D- 8ALACTOPYR ANOSYL - PINITOL
Figure 6.
St~ucture of ga1actopinito1.
Source: Schweizer et ~., 1978.

---

55
from non-fermented soya bean identified as melibiose in
the literature were in fact a galactopinitol.
Although the incidence of flatulence in humans is
unpredictable - depending on the psychological and physical
state of the subjects and the type of diet. oligosaccharides
are generally considered undesirable and many attempts have
been made by food scientists to treat beans or their
products to remove or degrade them.
Mital and Steinkraus
(1975) attempted to degrade oligosaccharides with lactic
acid bacteria.
Sugimoto and Van Buren (1970) used commer-
cial 8-galactosidase to hydrolyze oligosaccharides to their
component sugars.
Thanamunkul et ~. (1976) and Smiley
~~. (1976) used 8-galactosidase in a hollow fiber
reactor to hydrolyze oligosaccharides with varying degree
of success.
Rackis ~~. (1970) and Steggerda (1967),
working on experiment ~ vivo and in vitro, suggested that
antibodies and certain phenolic acids can inhibit flatus
activity.
Many workers have investigated the removal of oligo-
saccharides during soaking and germination (East et ~.,
1972; Hand, 1967; Iyengar and Kulkarni, 1977; Kawamura,
1966; Kim ~ ~., 1973; Ku ~~., 1976; Rao and Balavady,
1978; Reddy ~~., 1980).
Applications of ultrafiltra-
tion techniques in the removal of oligosaccharides had also
been used (Omosaiyre et !l., 1978).

---

56
Bean breeders are also attempting to identify genetic
variants possessing low levels of oligasaccharides which
may be used to biologically remove the flatulence factors
of beans.
Hymowitz et ~. (1972) have indicated that the
removal of oligosaccharides by plant breeding does not look
promising.
But because removal of oligosaccharides by
1) soaking results in loss of water soluble vitamin,
minerals and digestible sugars, 2) germination shows altera-
tion of the carbohydrate content of the seeds (Bond and
Glass, 1963; Linko ~~., 1960; Dubois ~ 2..!.., 1956),
3) chemicals need economic feasibility and approval for
human use, the general potential for eliminating flatulence
factors still lies in both biological and technological
areas.
Bean breeders can therefore still include that type
of research in their programs.
The quantitative determination of oligosaccharides has
been the subject of many investigations using either paper
chromatography, thin layer chromatography, column chro-
matography or gel filtration (Kawamura, 1967a, 1967b;
Hardinge ~ ~., 1965; De Stafanis and Gonte, 1968).
Delente and Ladenburg (1972) used gas-liquid chromatography
to quantitate oligosaccharides in defatted soybean meal.
Kim ~.!l..
(1973) employed liquid chromatography for. the
rapid determination of monosaccharides, disaccharides,
stachyose and raffinose in soybeans, but absolute values
for the quantity of the different sugars were not obtained

---

57
by this method.
Takana ~~. (1975) used an analytical
procedure based on thiobarbituric acid reaction (Percheron,
1962) to determine sucrose, raffinose and stachyose in
whole legume seeds.
All of the se methods are time consuming
and require sorne skill and experience to produce reliable
results.
Newly developed column packings for HPLC in the
recent years have greatly simplified sugar analysis in
foods and other plant materials.
Quality Evaluat~on of Processed Products
In the early history of breeding, the plant breeder who
was the cereal breeder and particularly the wheat-breeder
himself made his selections on the basis of agronomic
performance and disease resistance.
The main quality test
that he used was the II c hewing test ll , usually applied out in
the plots after hand threshing the grain of two or three
heads.
To an experienced person, the chewing test gave all
the quality information needed for early generation selec-
tion.
The pressure applied by the jaws to crack the wheat
gave a good, albeit subjective, measure of grain hardness.
As the .crushed grain was masticated in the mouth into a
dough, the starch and other solubles, due to salivary
amylase, were gradually dissolved and swallowed, or spit
out, depending on the number of samples tested.
Experience
showed that the size of the remaining gluten ball was indi-
cative of both protein content and gluten quality (Bushuk,

---

58
1982).
Gradually, as the cereal chemist's knowledge of the
fundamentals of milling and baking quality expanded,
screening of varieties for quality became much more sophis-
ticated.
Today as many as 26 different quality tests are
applied to material in the final stages of development
(Bushuk, 1982).
Those tests are applied now not only to
cereals but to many other food materials such as tubers,
roots and legumes.
Legume quality evaluation has been of
interest in the recent years by bean breeders because of
its role as a high protein source and supplement to cereal
products.
Adams and Bedford (1975) stated that the bases
for selection of improved breeding lines require additional
expenditures to conduct the various quality tests on the
canned product besides the quality tests on the dry seeds.
They mentioned the more subjective evaluations of quality:
1)
Wholeness - the tendency of the legume seed to
remain whole throughout the processing operations
not to break àpart, burst, or disintegrate.
2)
Consistency - the fluid is slightly viscous and
clear, not cloudy or grainy; it separates or drains
readily from the beans.
3)
Freedom from defects - no extraneous material,
loose skins, or mashed beans.
4)
Flavor ~ must be scored by a taste panel.

---

59
5)
Color - pigments in the seed coat that escape
detection in the dry seed may impart an off-color
to the cooked product.
6)
Texture - the measure of firmness, gumminess and
adhesiveness evaluated with reasonable accuracy by
a sensory panel and by machines such as Kramer Shear
Press, Instron and Ottawa Texture Measuring System.
Adams and Bedford (1975) stated also that there are no
comprehensive studies on the source and nature of variation
in the quality factors referred to above.
From limited
studies of special factors such as t~xture, and on the basis
of experience, the assumption has emerged that at least a
portion of the variability observed among lines depends on
genetic differences although it is clear that the length of
processing, the temperature of soak, the hardness of the
water, and the character of the added fluid all play an
important role.
The above two authors recognize that, in
practice, in order to assure acceptance of a new selection
by growers, the breeder generally will find it necessary to
compromise between the best level of quality possible in a
particular program and the levels of disease resistance and
agronomic performance that must be maintained.
They are
right to say that there is no escape from that because
consumer choice for a food product is primarily good
aesthetic appearance and qualities of the prepared food
except that now he or she becomes more aware of nutritional

---

60
evaluation of the food.
Hence quality evaluation of
processed foods becomes progressively important to plant
breeders in 0rder to meet the consumer requests and provide
nutritious foods with great acceptability.

---

MATERIALS AND METHODS
Sampl e' Source
Dry beans (Phaseolus vulgaris L.) used in this study
were: Sanilac, a standard commercial bean; Nep-2 (Nuclear
Experimental Project-2), a mutant; and San-Fernando, a
tropical genotype (Phaseolus vulgaris L.).
They were
obtained from Saginaw Valley Bean and Sugar Beet Research
Farm near Saginaw, Michigan, through the Department of Crop
and Soil Science, Michigan State University, East Lansing.
by the courtesy of Dr. G.L. Hosfield.
They were delivered
in paper bags to the Food Science Building and stored at
room temperature in the laboratory where the experiments
were carried out.
Seed Coat Separation
A certain amount of the seed from each type of bean was
immersed in warm tap water for about 5 minutes.
Coats were
then carefully removed because they were still attached to
the cotyledon outer sides.
The soaking period was short
in order to avoid diffusion of cell constituents such as
sugars into the water.
Each separated fraction was left at
room temperature for three days to dry.
61

---

62
Grinding of Samp1e
Cyclone Samp1e Mill, U.D. Corporation, was used to grind
into f1our, who1e seed, cotyledon (coat1ess) and coat from
each type of bean.
Moisture
Moisture was determined by oyen drying the samp1e at
80 + 20C unti1 weight remained constant in order to avoid
browning or carame1ization reactions taking place when
drying at 130 0C for 1 hr (Hosfie1d and Uebersax, 19 80 ).
Crude Fat
Crude fat was determined using A.D.A.C. method (1975)
based on extracting the fat from the bean f10ur with petro-
1eum ether in a Go1dfisch extractor.
Ash
The A.O.A.C. method (1975) was used for ash determina-
tion.
The method invo1ved oxidizing a11 organic matter in
a weighed samp1e by incineration at 550 0C unti1 the ash was
gray-white and determining the weight of the ash remaining.
Protein
The Kje1dah1 semi-micro-method based upon the determi-
nation of the amount of reduced nitrogen present was used.

---

63
The method consisted of: first, the wet oxidation of the
samp1e and the conversion of protein nitrogen into ammonium
sulfate; second, the decomposition df the ammonium sulfate
with 10 N sodium hydroxide and the distillation of the
ammonia evo1ved into saturated borie acid solution, and
fina11y, the determination of the nitrogen content by titra-
ting the ammonia with standard hydroch1oric acid, 0.1 N.
Sugars
Extraction and Injection
The simple and oligosaccharide sugars (glucose, sucrose,
inosito1, raffinose and stachyose) common1y found in beans
(Phaseo1us vu1garis L.) weredetermined using High Pressure
Liquid Chromatography (HPLC).
The method consisted of: first, an extraction of the
sugars in a 80% ethano1 :water (v/v) using a technique
deve10ped in our 1aboratory (Figure 7); second, precipi-
tation and centrifugation of protein materia1s by a solution
of 1ead acetate (10%, w/v); third, precipitation of excess
1ead in the extract using an oxa1ic acid solution (10%,
w/v), and remova1 of the 1ead oxalate by centrifugation
fourth, bringing-up of extract to volume in a 25 ml volu-
metrie f1ask; fifth, filtration of the prepared samp1e
extract through Waters Associates Sep-Pak C18 cartridges
having the same separation properties as the separation
co1umn C18 where the polar compounds e1ute before the

---

1
le
Add 10 ml 80~ ethano1
0
Shake in water bath at 80 C
for 15 min.
(1)
Sample f.11xture
l Centrifuge at 2000 RPM for 3 min.
Resi~ue Il
super'iatant Il
i
~
add 5 ml
''-'"'S'''u-p-e-r-n-a"T"t-a-n"T"t'''''
80% ethanol
and proceed as in (l)
~ 1a + b + C
Residue b
b~ ~I
1 Add 2 ml
Supernatant
~ 1
lead acetate
i
0'\\
.j::>o
(2)
l.fixture
Add 10 ml
80% ethanol
and proceed as in (1)
Centrifuge 3 min
at 2000 RPM
Resiilue c
Super~natant c
Ruperr.atant 2a
Residue 2
,
Stored for
add 2 ml
Star ch analysis
discard
oxalic acid
Supo~atant. 2b
itesidue
j
bring to
discard
volurne witt. H 0
Inject
Prefilter vith
1in 25 ml volu~etric
to HPLC ~(---- C
Sep-Pak
<
flask
18
Figure 7.
Flow diagram of sugar analysis.

---

65
non-polar ones.
Finally, the clear extract sample was
injected ante the C18 carbohydrate column (30 cm x 3.0 mm
1.0. plates/column N/A 3000, Waters Associates, Milford,
MA) for quantitation.
The elution solvent was filtered
acetonitrile:water (70:30, v/v, pH = 4.0) with a pump rate
of 2.0 ml/min.
Thirty microliter samples of the extracts
were injected using a 100 ~l pressure-lock microsyringe
into a model U6K injector which allows the sample to be put
on a bypassed injection part (no pressure exists in the
bypassed state).
After injection of the sample into the
U6K injector loop, the bypass valve on the Waters Associ-
ates pump system Model 6000A was then switched to introduce
the sample to the pressurized column held at room tempera-
ture.
The refractive index detector Waters Associates
Model R401 was used.
Methods of Quantitation
External standard and repetitive injection techniques
(Waters Associates 1980) were used.
They consisted of:
first, preparing a mixed standard solution containing a
known amount (1 mg/ml) of sugars to be quantitated; second,
injecting a sample of the standard solution to find out the
retention times of each individual known sugar appearing on
the peaks of a chromatogram through a Data Module computer-
ized machine (Waters Associates, Milford, MA); third, the
retention times of the sugars were put in calibration into

---

66
th~ computer Data Module according to the techniques of
Waters Associates (1980); fourth, the unknown samples
injected ante the HPLC column were then quantitated by
comparison with the standard sugars put in calibration
previously.
The percentage of the individual sugar extracted from
the beans used (Sanilac, Nep-2 and San-Fernando) were cal-
culated according to the following formula.
Percentage sugar =
Amount shown on data
x original sample
module (mg/ml)
value (ml)
x 100
sample weight (mg)
Starch
A new technique using HPLC was developed (Figure 8) and
used in this determination.
The technique consisted of
a) solubilization of the sample; b) hydrolysis of the
sample with amyloglucosidase enzyme to obtain glucose mole-
cules; c) injection of the hydrolysis product to the HPLC.
The amount of glucose obtained is relative to the amount of
starch hydrolyzed.
Sample Preparation
Reagents
sodium hydroxide (NaOH)
0.50 N
acetic acid
0.50 N

---

67
Starch Pellet
(~l.OOO g)
~
Disperse with 0.50 N NaOH (10 ml)
!
Neutralize with 0.50 N acetic acid (10 ml)
!
Neutral colloidal suspensions
Let stand few min. (not necessary)
L
Aliquot A
(1
ml)
Starch-Glucose assay
!
Add 3 ml amyloglucosidase (5 mg/ml)
~
Incubate 30 min/55°
20 C
continuous shaking
t
Remove tubes
!
Let cool (room temperature)
Filter throtgh amberlite
!
Injection to HPLC
Figure 8.
Flow diagram of starch dispersion, hydrolysis
and analysis.

---

68
The samples were the dry residue pellets obtained from
extraction techniques developed in this same laboratory.
10 ml of 0.50 N sodium hydroxide was added to the centri-
fuge tube containing the pellet (1.000 g).
With a rod, the
pellets were colloidally dispersed into the 10 ml NaOH
solutions (0.50 N) by continuous crushing and stirring.
After all the pellet was completely dispersed, 10 ml of
acetic acid solution (0.50 N) were added into the tube to
neutralize the sodium hydroxide used for dispersion.
The neutral colloidal suspensions were allowed to stand
for a few minutes in order to allow unsolubilized particles,
mainly cellulose, and pectic substances, to settle before
using the sample for injection (this step is not obligatory).
Buffer and Enzyme'Solutions
0.2 M Acetate Buffer Stock Solution
0.2 M acid:
12 ml glacial acetic acid was trans-
ferred to 1000 ml volumetric flask and diluted to
volume with distilled water.
0.2 M Sodium Acetate:
16.408 9 of ahhydrous
sodium acetate were dissolved in 800 ml of distilled
water, transferred to a 1000 ml volumetric flask
and diluted to volume.
Stock Solution:
Three volumes of 0.2 M sodium
acetate with two volumes of 0.2 M acetic acid were
mixed.
Volumetric flasks were used.
The pH was

---

69
4.9 (theoretica11y 4.92).
The solution was stored
in a we11-stoppered co1ored bott1e un der refrigera-
tion; it cou1d be stable for at 1east a month.
0.02 M Acetate Buffer Working Solution
To one volume of the 0.2 M acetate buffer stock nine
volumes of disti11ed water were added and shaken to
mix.
This dilution was carried out using pipette and
vo1umetric f1ask and prepared each time immediate1y
before each ana1ysis.
Enzyme Solutions
5 mg/ml amy1og1ucosidase (500 mg/100 ml, w/v) solutions
were prepared in 0.02 M acetate buffer working solution.
Enzyme Characteristics
Amy1og1ucosidase (glucoamy1ase 1,4-a-D-g1ucan gluco-
hydrolase; EC 3.2.1.3 from rhizopus genus mo1d 10,000 units/
9 solids).
The enzyme was purchased from Sigma Chemica1 Company,
St. Louis, MO.
Hydro1ysis
1 ml of samp1e was mixed with 3 ml of the enzyme amy1o-
glucosidase (5 mg/ml) solution in a test tube.
The samp1es
were p1aced in a water bath at 55 ± 20 C (optimum temperature
for amy1og1ucosidase) and continuous1y shaken for 30 min.
At the end of the time period, the test tubes were removed
from the water bath and fi1tered through a Sep-pak to

---

70
rem ove the acetate (Lester, 1980) prior to the injection to
the HPLC co1umn.
Injection
Injections proceeded as described above for the intro-
duction of sugars ante the C
carbohydrate co1umn.
18
Methods of Quantitation
Same as in the above quantitation of sugars with excep-
tion that on1y glucose was used as standard solution.
Ca1cu1ation
The percentage of starch was ca1cu1ated according to
the fo11owing formula:
Percent starch =
Amount shown on data
Original
module (mg/ml)
x 10 x vo 1ume (ml) x 100 x 0.9
samp1e weight (mg)
where:
0.9 (factor to account for the water gained during
hyd ro 1ys i s)
10 (equation correction factor)
Trip1icate determinations were performed on samp1es for
a11 above methods.

---

71
,
Water Absorption in Beans
The hydration properties of beans were determined by
soaking a 10 9 sample of dry beans (Sanilac, Nep-2 and
San-Fernando) contained in cylindrical grids in tap:dis-
tilled water (111) at room temperature for 0, 15, 10, 45,
60, 75, and 90 minutes.
After soaking, the cylindrical
grids were removed from the water and allowed to stand on
laboratory paper towel in order to remove surface water.
The beans were then weighed and the increase in weight
taken as the amount of water absorbed.
Triplicate deter-
minations were performed.
Water content was calculated using the following formula
(Hosfield and Uebersax, 1980).
100 _ (wt. of solids (g) in dry beans
wt.
(g) of soaked beans
) x 100
Canning
Beans used for canning evaluations were adjusted rapidly
to 16.0 ± 0.02% moisture content in a controlled air circu-
lating humidity chamber prior to soaking and processing.
This was done in order to eliminate any effect differential
seed moi sture might have on cotyledonary tenderization
during soaking and cooking and to insure that each sample
lot of beans contained a constant level of total solids
(TS).

---

72
The soaking and processing procedures developed by
Hosfield and Uebersax (1980) were applied in this process
(Figure 9).
The parameters considered in the evaluation of
the beans used in this experiment were: color of dry and
canned beans by Hunter colorimeter; drained weight by
decanting contents of cans on a number 8 mesh sieve,
rinsing in 21 0 C tap water to remove adhering brine and
0
draining for 2 minutes on the sieve positioned at a 15
angle; texture using a Kramer Shear Press (KSP) fitted with
a standard multiblade shear compression cell No. C338 (Food
Technology Corp., Reston, VA).
One hundred grams of washed
processed beans were placed in the compression cell and
force was applied until blades passed through the bean
sample while the instrument was set at range 10 (300 lb of
force full scale).
Water content of canned beans (final
moisture) was determined from 100 gm texture samples.
These were oyen dried at 80 0 ± 20 C to a constant weight
(Table 3).
Subjective bean quality evaluation was made on contents
of all processed cans while beans were drained on the mesh
screens.
The degree of packing (clumping) was rated on a
3 point scale.
Overall bean appearance was evaluated to
measure the suitability of beans to commercial processing.
Criteria included examining beans for loose or free coats
(llfree skins ll ), individual bean integrity, and fluid con-
sistency (Table 3).

---

5TORED DRY CEANS
~Q~KlliL'ti.uœ
30~ li 405
.BRINE FORMULA
-.....J
50 PPfA CA'"
CA"'
20 L1l WATER
W
CONDIllQli3
5 oz SUGAR
75 !lF/30 ""N~
4 O! SALT
190°F/30r.t1l4.
WATER 15 NIN.
240·F/4SMIN.
ISO-'=-
CANNED BEANS
Figure 9.
Schematic diagram unit operations for dry bean processing procedures.

---

74
Table 3.
Bean processing evaluation and calculations.
Character
Description
Soakinq
Hydration Coefficient
Ratio of wt of soaked beans (g)
wt of dry beans (g)
Water content (%)
100-
(wt of solids (g) in dry beans 100)
wt (g) of soaked beans
x
Canning
Objective
Drained weight (g)
Weight of rinsed beans drained
for 2 minutes on a number a mesh
screen (0.239 cm) positioned at
a 15 0 angle.
One determination
per can was made.
Texture (Kg force/100 g) Determined by placing 100 9 of
washed processed beans into a
standard shear-compression cell
of a Kramer Shear Press and
applying force with a dynamic
hydraulic system.
Values repor-
ted indicate the Kg force
required to sheat 100 9 of beans.
Three determinations per can were
made.
Water content (%)
Determined by oven drying each
texture evaluation at ao± 20 C
until weight remained constant.
%= initial wt - dry wt x 100
in i t i al wt
Subjective
Degree of packing (1-3)
Extent of packing (clumping) of
beans in cano
l = no clumping;
2 = bean clumping but easily
decanted from can; 3 = beans
clumped or packed solidly in
bottom of cano
One determination
per can was made.

---

75
Table 3.
(cont'd.).
Overall Appearance (1-5) Evaluation for general suita-
bility of commercial processing
made on each cano
Criteria
included examination for loose
(free) seed coats (split),
bean integrity, and brine con-
sistency.
Low values indicate
paor appearance; high values
indicate excellent appearance.
Source: Hosfield, G.L. and Uebersax, M,A.
1980.

---

76
A11 data were subjected to an ana1ysis of variance
appropriate to a comp1ete1y randomized design.
Dup1icate
readings were taken on a11 cans for texture and final
moisture content.
Texture Measurement Based on Single Bean
To accomp1ish this, sorne samp1es of our bean varieties
under investigation were sent to Dr. L.W. Hudson at the
Regional Plant Introduction Station, Washington State Uni-
versity, Pullman, Washington.
The tests were performed
using the apparatus deve10ped at that station.
The apparatus consists of 100 tests points in a la x la
array.
Each point has a cup holding a single bean seed.
Resting on each bean is a 1.47 mm diameter pin attached to
the end of a 1ength of tubing.
The tube contains enough
No. 8 shot so each of the 100 pins bears 90.6 grams on each
bean.
The cooking vesse1 contains about 180 1iters of
water heated by steam-coi1s to 93.3 0C.
A Taylor contro11er
0
keeps the water at a very constant temperature (± 0.25 C).
When the water is at the prescribed temperature, the testing
device is put in the water and the time recorded as "Beans
in".
Simu1taneous1y a timer is started and in due course,
as each bean reaches the proper state of "cook" the pin
~
penetrates and the time is noted in the square corresponding
to the test point.

---

77
The conditions of the test were: a) the moisture content
of the seeds was adjusted by humidifying the seeds for 6
hrs; b) the beans were scarified by clipping away a small
portion of the pericarp on the edge of the seed opposite
the hilum and then c) soaked for 12 hours using distilled
water kept at room temperature.
Tests have shown that with the apparatus, significant
differences can be shown with ten seeds per lot.
In this
study, the test was performed four times using best looking
bean seeds (twice with seed coat intact and twice with seed
coat partially removed).
The results were reported as time
to breakdown (TTB) in minutes with standard deviation and
coefficient of variation.
Gelatinization Characteristics
A sample of 50 9 of whole bean and cotyledon flour
(adjusted to 14% moisture) fram each type of bean was used
with 400 ml of phosphate buffer, pH 5.30 to make the slurry
ready for heating.
The functional
properties of the flours
were evaluated according to the AACC methods using Braben-
der Amylograph (1969).
The method of graphic analysis for
functional properties are as follows (Table 4) (Mazurs
~ ~., 1957; Kite ~~., 1957):
The Peak Viscosity or Pasting Peak
This is the highest viscosity which is reached during
the gelatinization of the starch.
The temperature where

---

Table 4.
Functional and molecular properties associated with pasting characteristics of
starches.
Paste Properties
Functional Properties
Molecular Properties
(Experimentally Determined)
Rate of increase in viscosity
Ease of cooking
Rate of granules swelling
when heated to 9S oC
(Region prior to Point A)
Viscosity peak (Point A)
Maximum thickness on
Extent of granule swelling
cooking
Viscosity changes (after
Stability during cooking
Granule fragility and degree
reaching maximum viscosity)
of solubilization
during heating and 9S oC
""'-J
00
holding cycles (region of
Points A to C)
Increase in viscosity during
Set-back on cooling
Regragradation of linear
cooling (region of Points C
molecules
to 0)
Changes in viscosity during
Resistance to shear
Granule rigidity
holding at saoc (region of
Points 0 to E)
Final viscosity after holding
Thickening power or
Granule rigidity extent of
at saoc (Point E)
thickening efficiency
maintained swelling
Source: Mazurs ~~., 19S7.

---

79
the viscosity begins to increase, and the rate of increase
are also considered.
Together these three factors indicate
the ease of cooking and the pasting peak provides an esti-
mation of the power requirements for stirring the starch
paste during gelatinization.
Sorne starches do not have a distinct peak.
The visco-
sity simply increases during heating and tends to remain
relatively constant during the holding cycle at 95 0 C.
The Viscosity at the End of the Heating Cycle as Sample
Reaches 95 0 C
This gives an indication of stability during cooking
when related to peak viscosity.
A sharp drop in viscosity
from the viscosity peak indicates granule fragility and
solubil ization.
The Viscosity at the End of the 95 0 C Holding Cycle
This indicates the degree of fragility or stability of
the hot paste.
A drop suggests additional breakdown of
granules or solubilization due to stirring.
The Viscosity at the End of the Cooling Cycle, When the
Paste Reaches Again 50 0 C
This is a measure of the thickening or " se t-back" of
the paste when cooling.
It arises from retrogradation of
the linear molecules and is a serious obstacle during pro-
cessing.

---

80
The Viscosity at the End of the 50 0 C Holding Cycle
This indicates the stability of the paste to stirring
in the form in which it will most likely be used by the
industry.
It is a good indication of granule rigidity and
resistance to shear.
The actual viscosity at this point may
also be considered as a measure of thickening power or
thickening efficiency of starch.
Table 4 summarizes the relationships between the Bra-
bender viscosity curves and the functional
properties.
These functional
properties are the basis for determining
the usefulness of a good starch.
Table 4 also indicates
the molecular events which are believed to be responsible
for the observed changes (Mazurs ~ ~., 1957; Carson, 1972).
Medcalf and Gilles (1966) and Schoch and Maywald (1968)
also described the terminology used to express the amylogram
results.
The temperature of initial pasting for this work
is defined as the temperature at which the viscosity curve
starts rising during the heating period.
Perpendicular
rising temperature is the temperature at which the C-shape
curve starts rising and a peak reading is taken just at the
top of that first curving point.
The normal peak height
is taken at the second curving point of the C-shape curve.
In the cases where a definite sharp peak is not obtained,
no value is given because height values at 95 0 C are repor-
ted.
The peak after 15 min is the viscosity of the sample
after 15 min holding period at 95 0 C.
The peak drop is the

---

81
height obtained at the drop point of the curve during the
cooling period.
The height is the viscosity of the sample
0
after it has cooled to 25 C.
Peak on cooling is the height
of the curve obtained during the 15 min cooling period.
0
After 15 min at 25 C is the height reached after the 15 min
cooling period.
Scanning Electron Microscopy
Dehydration
Sean samples (dry, soaked, canned) were freeze-dried
in a freeze dryer Unitrap II (Virtis, Gardiner, NY, 12525),
4 to 8 hrs respectively.
Coating and Viewing
Dried beans were dry-fractured by hand or using blade
to 1I 0pen ll tissues and cells, mounted on stubs with colloidal
finger polish and coated with an approximately 20 nm layer
of gold using a sputter coater.
The coated samples were
viewed and photographed in a Philips Super III Scanning
Electron Microscope (SEM) at an acceleration voltage of 15
Kv with a Polaroid PIN film type 665.

---

82
Freeze-Orying
~
Ory-fracturing
~
Coating
~
Viewing
Figure 10.
Flow diagram of sample preparation for SEM.
Statistical Analysis
The IIStatistical Package for Social Science ll computer
programs described by Nie et al. (1975) for use on the COC
6500 computer operated by Michigan State University Computer
Laboratory was used ta assist statistical analyses.
Multivariate analyses of variance and covariance were
determined using subprogram ANOVA.
Mean values were repor-
ted after roundings.
Single classification analyses of
variance, Tukey mean separations were determined using
subprogram ONEWAY.
Tukey separations were presented such that treatments
which were not significantly different Cp?-0.05) were indi-
cated with like letters.
Sean quality parameters and quality evaluation were
subjected ta an analysis of variance appropriate ta a
completely randomized design.
Mean squares with significant
F ratios were used ta determine significant probability
level of p 0.05.

---

RESULTS AND DISCUSSION
Proximate Composition
The proximate composition of the three dry bean cultivar,
Sanilac, Nep-2 and San-Fernando over three crop years is
shown in Tables 5, 6, 7, 8, 9, 10, 11, 12,13 and 14.
Moisture content was higher in the 1978 beans than in
the other two crop years (Table 5).
Few significant dif-
ferences were obtained within the crop years among the
three cultivars of whole bean.
The combined result indi-
cated no significant differences among the cultivars for
the three portions of the seed.
The seeds have similar high crude protein contents as
obtained in other Phaseolus vulgaris L. beans (Ruth ~ ~.,
1979; Pedro and Ladermiro, 1979; Shalini et ~., 1968;
Valdemiro ~ ~., 1979).
There were no significant dif-
ferences in the whole bean and seed coat protein for the
three cultivars over the three crop years except San-Fer-
nando which showed significant differences for
the 1979
and 1980 beans (Table 7).
In cotyledon alone no singificant
differences were observed among the varieties except Nep-2
and Sanilac cotyledons which possessed significant differ-
ences in the 1978 and 1980 beans, respectively.
The
83

---

Table 5.
Percent moisture content (dry basis) of bean cultivars (Sanilac, Nep-2 and
San-Fernando) by crop years (1978, 1979 and 1980) and seed portions (whole bean,
cotyledon and seed coats.
Crop Years
-
1978
1979
1980
Cultivars
Seed Portions
Seed Portions
Seed Portions
Whole
Cotyledon Seed
Whole
Cotyledon
Seed
Whole
Cotyledon
Seed
Coat
Coat
Coat
a
Sanilac
8.7
7.2 b
8.3 e
6.8 a
6.3 b
6.5 c
6.3 a
5.9 b
5.4 d
c
Nep-2
8.7 a
9.2
8.3 e
6.7 a
6.2 b
6.7 c
6.3 a
6.0 b
5.2 d
d
SF
8.6 a
9.9
9.0 f
6.6 a
6.3 b
7.3 d
6.5 a
5.3 c
5.6 d
00
.j::>o
n = 3 (3 replicates/seed portion/cultivar.
Like letters in column denote non-significant differences (p~0.05) .among cultivars and
within crop years and seed portions.

---

85
Table 6.
Percent moisture content (dry basis) of bean
cultivars (Sanilac, Nep-2, San-Fernando) for
three crop years (1978, 1979 and 1980) by seed
portions (whole bean, cotyledon and seed coat).
Seed Portions
Cultivars
Whole
Cotyledon
Seed Coat
a
b
c
Sanilac
7.3
6.5
6.8
b
c
Nep-2
7.2 a
7.2
6.7
a
b
c
SF
7.2
7.2
7.3
n = 9 (3 repl icates/seed portion/cultivar x 3 years).
Like letters in column denote non-significant differences
(p'0.05) among cultivars and within seed portions.

---

Table 7.
Percent protein content (dry basis) of bean cultivars (Sanilac, Nep-2 and
San-Fernando) by crop years (1978, 1979 and 1980) and seed portions (whole bean,
cotyledon and seed coat).
n = 9 (3 replicates/seed portion/cultivar x 3 injections/replicate).
Like letters in column denote nonsignificant differences (p~0.05) among cultivars and,
within crop years and seed portions.

---

87
Table 8.
Percent protein content (dry basis) of bean
cultivars (Sanilac
Nep-2 and San-Fernando) for
t
three crop years (1978
1979 and 1980) by seed
t
portions (whole bean
cotyledon and seed coat).
t
Seed Portions
Cultivars
Whole
Cotyledon
Seed Coat
b
Sanilac
24.0 a
25.8
7 . 7d
a
d
Nep-2
23.8
23.3 c
7.5
d
SF
23.8 a
24.7 bc
6.7
n = 9 (3 replicates/seed portion/cultivar x 3 years).
Like letters denote nonsignificant differences (p~0.05
among cultivars and within seed portions.

---

Table 9.
Percent fat content (dry basis) of bean cultivars (Sanilac
Nep-2 and
t
San-Fernando) by crop years (1978
1979 and 1980) and seed portions (whole bean
t
t
cotyledon and seed coat)
Crop Years
1978
1979
1980
Cultivars
Seed Portions
Seed Portions
Seed Portions
Whole
Cotyledon
Seed
Whole
Cotyledon
Seed
Whole
Cotyledon
Seed
Coat
Coat
Coat
b
c
e
d
Sanilac
l .3 a
1. 7
0.8
l .4 a
l .4 c
0.4
l . l a
1. l b
O.5
b
c
d
f
d
Nep-2
l .6 a
1. 8
0.6
l .5 a
1. 8
0.6
l .2 a
l .4 c
0.5
00
cd
b
e
l .2 a
1. 7 b
e
00
SF
0.7
1. 2
l .9 d
0.4
l .2 a
1. lb
0.2
n = 3 (3 replicates/seed portion/cultivar).
Like letters in column denote nonsignificant differences (p~0.05) among cultivars and t
within years and seed portions.

---

89
Table 10.
Percent fat content (dry basis) of bean
cultivars (Sanilac, Nep-2 and San-Fernando) for
three crop years (1978, 1979 and 1980) by seed
portions (whole bean, cotyledon and seed coat).
Seed Portions
Cultivars
Î'lho le
Cotyledon
Seed Coat
Sanilac
l .3 a b
l .4 c
0.6 d
Nep-2
l .4 b
l . 7c
O.Sd
SF
l .2 a
l .6 c
0.4 d
n = 9 (e replicates/seed portion/cultivar x 3 years).
Like letters in column denote nonsignificant differences
(p)O.OS) among cultivars and within seed portions.

---

Table 11.
Percent ash content (dry basis) of bean cultivars (Sanilac, Nep-2 and
San-Fernando) by crop years (1978, 1979 arld 1980) and seed portions (whole
bean; cotyledon and seed coat).
Crop Years
-
1978
1979
1980
Cultivars
Seed Portions
Seed Portions
Seed Portions
Whole
Cotyledon
Seed
Whole
Cotyledon
Seed
Whole
Cotyledon
Seed
Coat
Coat
Coat
a
c
e
a
c
e
Sanilac
5.2
4.0
6 . l
4. l
3.9
6.7
4.9 a
4.6 c
6. l f
b
d
f
b
cd
f
4.0
4.7
4.5 b
4.4 d
5.2 f
Nep-2
4.2
4 . l
5.4
3.9
b
g
a
I.D
c
3.9
4. l d
f
3.7
4.4 b
4.3 e
3.9 g
0
SF
4 . l
3.9
4. l
n = 3 (3 replicates/seed portion/cultivar)
Like letters in column denote nonsignificant differences (p~0.05) among cultivars and,
within crop years and seed portions.

---

91
Table 12.
Percent ash content (dry basis) of bean
cultivars (Sanilac, Nep-2 and San-Fernando) for
three crop years (1978, 1979 and 1980) by seed
portions (whole bean, cotyledon and seed coat).
Seed Portions
Cultivars
Whole
Cotyledon
Seed Coat
a
c
d
Sanilac
4.7
4.2
6.3
b
c
Nep-2
4.2
4.2
5. l e
b
f
SF
4.2
4. l c
3.8
n = 9 (3 replicates/seed portion/cultivar x 3 years)
Like numbers in column denote nonsignificant differences
(p~0.05) among cultivars and within seed portions.

---

Table 13.
Percent starch content (dry basis) of bean cultivars (Sanilac, Nep-2 and
San-Fernando) by crop years (1978, 1979 and 1980) and seed portions (whole
bean, cotyledon and seed coat).
Crop Years
1978
1979
1980
Cultivars
Seed Portions
Seed Portions
Seed Portions
Whole
Cotyledon
Seed
Whole
Cotyledon
Seed
Whole
Cotyledon
Seed
Coat
Coat
Coat
a
a
b
Sanilac
45.4
41 .6 c
4. l d
52.0 a
39.4 c
Trace
4S.S
47.0
Trace
ab
c
d
c
d
Nep-2
49.5
47.2
2.7 e
4 l . 7 b
SO.3 c
6.2
37.3 a
42.4
6.7
d
56.6 b
3.2 de
40.0 b
4.8
b
1.0
SF
48.7 c
43.2 c
47.9 a
47.7
Trace
N
n = 3 (3 replicates/seed portion/cultivars).
Like letters in column denote nonsignificant differences (p~O.OS) among cultivars and,
within crop years and seed portions.

---

93
Table 14.
Percent starch content (dry basis) of bean
cultivars (Sanilac, Nep-2 and San-Fernando) for
three crop years (1978, 1979 and 1980) by seed
portions (whole bean, cotyledon and seed coat).
Seed Portions
Cultivars
Whole
Cotyledon
Seed Coat
a
b
Sanilac
47.6
42.3
l .5 c
a
b
d
Nep-2
42.8
46.7
5.2
b
cd
SF
48. la
46.5
2.7
n = 27 (3 replicates/seed portion/cultivar x 3 injections
x 3 years).
Like letters in column denote nonsignificant difference
(p~0.05) among cultivars and within seed portions.

---

94
combined three crop years resu1ts showed significant dif-
ferences among the cultivars for cotyledon protein content
ranging from 23.3% (Nep-2) to 25.8 (Sani1ac (Table 8).
Fat content constitutes a re1ative1y sma11 amount of the
overa11 bean composition 1ike in most low fat 1egumes
(Table 9).
There were no significant differences in the
who1e seed fat content among the cultivars except that
San-Fernando showed a slight difference in the 1979 beans.
Significant differences in fat were obtained in the cotyle-
don from the 1979 and 1980 beans but not in those of 1978
(Table 9).
Seed coat fat content was 1ess than one percent.
Nep-2 and San-Fernando showed no significant differences in
their seed coat fat content for the year 1978.
Sani1ac and
Nep-2 showed no significant differences for 1980 beans (Table 9).
Combined fat content from the three crop years resu1ts
indicated significant differences in the who1e bean among
the cultivars but not in the beans cotyledons and seed
coats (Table 10).
Ash content fa11s into the range reported for 1egumes
(Phaseolus vulgaris L.) by Ruth et ~. (1979), Pedro and
Ladermiro (1979) (Table 8).
The ash content did not show a
significant difference between the two isolines 1978 and
1980 whole bean samp1es (Table 11).
Within the years,
significant differences were obtained for cotyledons and
seed coats among the three cultivars.
The combined percent
ash content revealed no significant differences between the

---

95
two isol ines in whole beans and cotyledons however, in the
seed coat a significant difference was obtained (Table 12).
The starch content in Table 13 is similar to other
legumes Phaseolus vulgaris L. reported by Navikul and
d'Appolonia (1978), Cerning and Aliette (1979), Roberto
and Eidiomar (1971), and Theravuthi ~~. (1974).
There
was no significant differences between the two isolines for
the three seed portions (whole, cotyledon and seed coat)
for the three crop years (Table 14), except San-Fernando
which showed sorne difference in the cotyledon with the 1980
beans.
In general, the statistical analysis of the proximate
composition revealed, except for fat content, no signifi-
cant differences for the protein, ash and starch contents
in the whole beans among the three cultivars although dif-
ferences were obtained in sorne cotyledon and seed coats.
In this case, we can not rely on the proximate composition
to explain the textural behavior differences of the beans
used in the experiment.
Sugars
Sugars commonly found in legumes (Phaseolus vulgaris L.)
are reported in Table 15A, B, and C.
Hexose represents a
mixture of D-glucose and D-galactose, and probably D-
fructose because in this study, these three reducing sugars
appear at the same retention time from a standard " coc ktail"

---

Table lS.
Percent sugar (hexose, sucrose, inositol, raffinose and stachyose) content (dry basis) of bean cultivars (Sanilac,
Nep-2 and San-Fernando) and seed portions (whole bean, cotyledon and seed coat) by crop year (1978, 1979 and 1980).
Hexose
Suc rose
Inositol
Raffinose
Sta chyose
Seed Portion
Seed Portion
Seed Portion
Seed Portion
Seed Portion
Whole
Cotyledon
Seed Wh"o 1e
Cotyledon
Seed Whole
Cotyledon
Seed Whole
Cotyledon
Seed Whole
Cotyledon
Seed
Coat
Coat
Coat
Coat
Coat
A. Crop Year 197a
a
b
c
b
b
b
a
Sanilac
.9
.9
.2
2.1 a
2.S
.Sc
.Sa
.6
• 1c
• Ja
.J
ND
1. 4
1. 1b
ND
a
d
a
b
Nep-2
.aa
.9 b
.1 d
2.J
2.8 b
.2
.4
.4 b
ND
• Ja
.4
ND
1. Sa
1. 1b
ND
a
c
a
b
cd
a
SF
.9
1. Ob
.2
2.0
2.S
.J
.4
.4 b
.lcd .Ja
.Sb
ND
1. 4a
1. 2b
ND
a. Crop Year 1979
a
d
Sanilac 1. 7
1. 4b
2.0 c
1.6a
2.9 b
.2 c
.7 a
.6
.1 f
• Ja
.Sb
ND
2.0 a
2.7 b
ND
a
Nep-2
1. 9
1.6 b
1.8c
1. aa
J.J b
.2 c
.2 b
.J de
II f
.4 d
.4 b
ND
2.0 a
J.D b
ND
U)
1. Jb
2.9 b
.1 d
.Sb
b
C"l
SF
2.J a
1. SC
2.0 a
.4 c
.2 e
ND
.sa
ND
2.2 a
2.a
ND
C. Crop Year 19aO
a
Sanilac
.aa
.a d
ND9
1. 9
2.2 c
.2 d
.6 a
.Sb
ND
• aa
.Sc
Nol 2.7 a
J.s d
ND
Nep-2
1. 1b
1. Je
.1 9
2.6 b
1. 9c
.2 d
• Sa
.4 b
ND
ND
ND
ND
J.O b
2.6 e
ND
SF
1.8 c
1. 9f
.1 9
2.2 ab
2.Jc
.2 d
.7 a
.ac
ND
.J b
• JC
ND
2.4 c
2.J f
ND
n ~ 9 (J replicates/seed portion/cultivar x J injections/replicate)
lND • Nondetectable
Like letters in column denote nonsignificant differences (p~O.05) among cultivars and within seed portions.

---

97
mixture of these sugars.
It has previously been reported
that anomeric forms of reducing sugars such as D-glucose
and D-galactose are not resolved in HPLC (Palmer. 1975;
Eileen et ~"
1979).
Two flatulent sugars raffinose and
stachyose, which are of concern to consumers. were eluted.
The stachyose content was higher as compared to raffinose.
In general, no significant differences for each sugar were
shown in the whole bean, cotyledon and seed coat of the
three cultivars for the three crop years (Appendix Table lA).
Hexose and sucrose did not show any significant differences
in the whole bean and cotyledon for the crop years 1978 and
1979 for the three cultivars.
The combined results were
similar although they differ in the 1980 bean samples
(Tables 15A, B, and Appendix Table lA).
Higher hexose
contents ranging from 1.7 to 2.3% were obtained in 1979
whole beans (Table 15B) and lower values in those of 1978
(Table 15A).
Stachyose showed its highest content (2.4%.
2.7% and 3.0%) in 1980 whole beans (Table 15C) and lowest
(1.4%, 1.4% and 1.5%) in 1978 beans (Table 15A).
Nep-2 and
Sanilac had the highest (3.0%) and lowest (1.4%) stachyose
values respectively (Table 15C and 15A).
The 1980 beans
revealed no detectable amount of raffinose in the entire
beans (Table 15C).
The inositol content was less than 0.7%
in the beans used and, in the three crop years combined
results, it did not show any significant difference at the
three portions among the isolines (Appendix Table lA).
The

---

98
seed coat present no detectable raffinose and stachyose.
Combined results are presented in Figures 11, 12 and 13.
All the results obtained on sugars fall
in the range of
values reported in legumes by Navikul and d'Appolonia
(1978), Quemener and Mercier (1980), Cerning ~~. (1975),
Akpapunam and Markakis (1979), and Munehiko ~~. (1975).
The variation in the content of the individual sugars
from year to year could be related to soil conditions,
weather, nutrient availability.
Nep-2,which showed no
detectable amount of raffinose in the 1980 whole beans,
possessed the highest value of stachyose (Table 15C).
This
implies a compensation process which could be important to
plant breeders in the selection of cultivars leading to
elimination of flatulent factors in beans.
Scanning Electron Microscopy Examination
of Dry Beans
Examination of the seed coat outer surface showed both
Sanilac and Nep-2 to possess a highly rough and wrinkled
outer surface with pores and sinking holes while San-Fer-
nando revealed only a highly rough and convoluted structure
(Figure 14A l , Bl , Cl).
The coat inner surface views showed
San il a c and Nep - 2 wi th sim i l a r structure, wide Il hi Ils Il and
narrow "valleys" (Figure 14A2, B2 , C2).
The seed coat of Sanilac was not difficult to remove
compared to the two isolines Nep-2 and San-Fernando.
A

---

NILAC
~ SA
3.00 ~
O.NE -2
N-FERNANDO
~ SA
-
~
2 00 ~
~
lO
.
1
....
~
lO
10:-
-~-
-~-
Io~-
m
-~-
10--
~~-
10--
~~-
10--
ffi
--.
10--
o
,
r.w:
-~.
~--
~ ! 1.00 -
-
-
.~-
:~:.
•-
-.
_.
•
.--
-
_.
••
-
·-
-
- -
•
10--
-
~
--
10--
-
Ie~-
~--'
-
-.
10--
-
r ;
-
~~-
Ie-- -
-
• -
-
---
-.
-
..
~~-
.~.
..
•
.~.
.~.
0.00
..'
•••
·0
•
~
•
• •
•••
• •
• •
1
tEXOSE
SUCROSE
INosrrOL
RAFRNOSE
STACHYOSE
Figure 11.
Mean sugars (hexose, sucrose, inositol, raffinose and stachyose) content values
over three crop years (1978, 1979 and 1980) for whole bean cultivars (Sanilac,
Nep-2 and San-Fernando).

---

mSANILAC
3.00 ..
DNEP-2
mSAN-FERNANDO
.-
':1
..-
..:~I--
~
• •
•
2.00 II'
• •
.....
•
• •
• •
•
•
• •
• •
•
•
• •
• •
•
•
• •
• •
•
•
• •
'"
• •
•
•
• •
• •
•
~
IX
•
• •
• •
•
o
• •
•
o
m '
1.00
,..., IX
•
• •
~
IX
•
• •
•
• •
•
x
IX
..'
•
• •
• •
•
IX
• •
•
• •
•
• •
•
• •
:fi
• •
•
x..
•
• •
•
o
• •
•
....
• •
•
• •
•
• •
•
•
Bi
~
• •
•
• •
•
•
•
•
•
• •
Do
•
• •
•
•••
•
•
•
•
• •
•
••
•
•••
•
..
•
• •
•
"
IX
•
•
•••
0.00
·
...
•
HEXOSE
SUCROSE
INOSrrOL
RAFFfiOSE
STACHYOSE
Figure 12.
Mean su gars (hexose, sucrose, inositol, raffinose and stachyose) content values
over three crop years (1978, 1979 and 1980) for bean cultivar (Sanilac, Nep-2
and San-Fernando) cotyledons.

---

3.001-
~SANILAC
DNEP-2
m SAN-FERNANDO
1
~ 2.00
CJ
:::»
0
!z
lU
0
0
1
.......
(fi
1.00
Do
o.oo~
ne!n
ND
ND
tEXOSE
SUCROSE
INOSITOL RAFFINOSE
STACHYOSE
Figure 13.
Mean sugar (hexose, sucrose, inositol, raffinose and stachyose) content values
over three crop years (1978, 1979 and 1980) for bean cultivars (Sanilac, Nep-2
and San-Fernando) seed coat.

---

102
NEP-2
..
~-
BAN-FERNANOO
Figure 14.
Scanning electron micrographs of dry bean seed
coat outer (1000x) and inner (1000x)
surfaces
structures; Al = Sanilac coat outer surface;
A2 = Sanilac coat inner surface; 81 = Nep-2
coat outer surface; 8 2 = Nep-2 coat inner
surface; Cl = San-Fernando coat outer surface;
C2 = San~Fernando coat inner surface.

---

103
sticky membrane frequently remained on the cotyledon of
Nep-2 and San-Fernando when the seed coats were removed.
A
larger membrane was always observed on the San-Fernando
cotyledon than on Nep-2 (Figure 15B l and Cl)'
Higher mag-
nification (Figures 15A 2 , B2 , C2 ) revealed Sanilac and
Nep-2 with similar structure compared to San-Fernando.
Contrary to th~ inner surface of the seed coat, cotyledon
surfaces were rough and covered by wide "hills" with narrow
Il val l e y s Il
(F i 9ure l 5A2' B2' C2 ), wh e r e a s the i nners ur f ace
of the seed coat was covered with narrow "hills" and wide
"valleys" (Figure 14A 2 , B2 , C2 ) as was reported by Sefa-
Dedeh for cowpea cultivar Adua Ayers (1978).
As reported
by Sefa-Dedeh (1978) in cowpeas, the surface topography
suggested also that Sanilac, Nep-2 and San-Fernando cotyle-
don surface and seed coat inner surface may be of comple-
mentary structures, the "hills" of the cotyledon fitting
into the "valleys" of the seed coat.
The protein membrane
found on the cotyledon surface of Nep-2 and San-Fernando
shows a tighter interlocking structure of the cotyledon
and seed coat in those two isolines.
This prevents a rapid
translocation of water between the cotyledon and the seed
coat; hence slow absorption of water into the parenchyma
cells of the cot~ledon of those isolines.
San-Fernando
cotyledon covered with a larger portion of the membrane
showed the slowest water absorption pattern.
In external topography, distinct hypocotyl area, elli.p-
tical hilùm, and micropyle are visibel at low magnification

---

104
8ANlLAC
8ANlLAC
..
NEP-2
SAN -FECI ..... 41\\100
Figure 15.
Scanning electron micrographs of 'dry bean seed
cotyledon outer surface structures (30x and
700x); Al and A2 = Sanilac cotyledon outer
surface; Bl and B2 = Nep-2 cotyledon outer
surface; Cl and C2 = San-Fernando outer surface.

---

105
in all three cultivars as commonly found in legumes (Figures
16A , B , Cl)'
Micropyle just below the hilum indicated
l
l
specific differences by being heart-shaped open, Y-shaped
open and Y-shaped closed for Sanilac, Nep-2 and San-Fernando,
respectively (Figure 16A2' B2 , C2).
Sanilac showed a flat hypocotyl area, Nep-2 a slightly
elevated one, and San-Fernando a highly elevated area
(Figure 17A l , B , Cl)'
This is a meaningful difference
l
among the three cultivars because all
individual seeds
from each cultivar showed this characteristic.
The hilum
structure consists of a double layer of palisade cells like
in soyabean reported by Kondo (1913) and Wolf and Baker
(1972); the upper layer gives the hilum a meshlike structure
(Figure 17A 2 , B2 , C2 ).
Double layers of palisade cells are
seen transversely through the hilum (Figure 17A , B
).
2
2 , C2
Immediately below the hilum groove are the tracheids which
are similar in all three cultivars except that those of San-
Fernando are narrower than the other two (Figures 17A , B ,
2
2
C2 ) .
The seed testa showed all
the major anatomical charac-
teristics of legume seeds as reported by Corner (1951).
Longitudinal and transverse cross-sections of the bean seed
coats show similar structure (Figure 18).
The palisade thickness were about 23 ~m, 30 ~m and 32 ~m
for Sanilac, Nep-2 and San-Fernando, respectively.
Only a
single layer of hour-glass cells was present and was thinner

---

1 06
NEP-2
M
SAN-FERNANDO
SAN-FERNANCO
Figure 16.
Scanning e1ectron micrographs of dry bean seed
hi1um area structures.
Al = Sani1ac hi1um area (50x); A2 = Sani1ac
mlcropy1e (1000x); B1 = Nep-2 hilum area (50x);
B2 = Nep-2 micropyle (lOOOx); Cl = San-Fernando
hl1um area (50x); C
=
2
San-Fernando micropy1e
(1000x); HC = hypocoty1 area (50x); H = hi1um;
M = micropy1e.

---

107
SANILAC
SANILAC
NEP-2
Figure 17.
Scanning electron micrographs of dry bean seed
hypocotyl area (100x) and hilum transverse
cross-section (200x) structures.
Al = Sanilac
hypacotyl area; A2 = Sanilac hilum transVerse
section; B1 = Nep-2 hypocotyl area; B2 = Nep-2
hilum transferse section; Cl = San-Fernando
hypocotyl
area; C2 = San~Fernando hilum
transverse section; HC = hypocotyl area; H =
hilum; T = tracheid

---

108
B ANILAC
BI..
.
, . ' .
.
. • _
•
Ct:.
.
. _..
_
,.
.. '
' .
' .
"
~
" ~
.-
SAN - FERNANDO
Figure 18.
Scanning electron micrographs of dry bean seed
coat cross-section structures (1600x).
Al = Sanilac longitudinal cross-section;
A2 = Sanilac transverse cross-section;
Bl = Nep~2 longitudinal cross-sect'ion;
B2 = Nep-2 transverse cross-section;
Cl = San-Fernando longitudinal cross-section;
C2 = San-Fernando transverse section; PAL =
Palisade cells; HGl = Hour-glass cells

---

109
than the palisade with thickness of 21
~m, 17 ~m and 13 ~m
for Sanilac, Nep-2 and San-Fernando, respectively.
Hour-
glass cells appear more organized and rigid in structure
for each bean cultivar.
San-Fernando has the thinner (13
~m) hour-glass and thicker (32 ~m) palisade size.
Ratio of
palisade to hour-glass indicates about 1.1, 1.8, and 2.5
fold thickness in Sanilac, Nep-2 and San-Fernando, respec-
tively.
This high palisade thickness could be a factor in
the slow absorption of water through the San-Fernando seed
coat compared to the other two cultivars.
The combined
palisade and hour-glass thicknesses were approximately the
same for the three cultivars by being
45 ~m in Sanilac,
47 ~m in Nep-2 and 45 ~m in San-Fernando.
Amorphous meso-
phyll layers were seen in all the cultivars next to the
hour-glass cells (Figure 18).
The presence of a single
layer of hour-glass cells has been reported in most bean
species by Chowdhury and Buth (1970).
Figure 19 presents the seed cotyledon inner side surface
structure of the three cultivars under investigation.
Nep-2
and San-Fernando cotyledon inner side surfaces are similar
in structure with well organized protein film bundles
covering the entire inner surface while Sanilac shows a
protein film with thinner bundles.
The effect of the film
bundles on the water absorption process is not known, how-
ever it may be a barrier to the easy penetration of water
into the cotyledon parenchyma cells.
The two isolines which

---

l l a
SANILAC
\\
1
J
1
, 1
1
1
i:,
,
Figure 19.
Scanning electron micrographs of dry bean seed
cotyledon in1e~ side surface structur~s (3ax
and 4aOx).
Al and AZ = Sanilac cotyledon inner
side surface; Bl and B2 = Nep-2 cotyledon
inner side.surfacc:; Cl and C2 = San-Fernando
----:-:--1
cotyledon lnner slde surface; HP = hypocotyl.
;

---

111
have different structure at the cotyledon outer side
surface show a genetic similarity in structure on this side.
of the cotyledon (Figure 198 2 , C ).
2
The 1980 Nep-2 beans revealed seed cotyledon naturally
divided in sections (Figure 20A,8, C).
This presentation
was different from Nep-2 seeds obtained from previous crop
years and also to Sanilac and San-Fernando seed cotyledons
in general.
The fracture lines seen clearly on the inner
side surface of the cotyledon (Figure 208, C) were observed
on most of the seeds of that crop year whîle no such pheno-
mena was seen in the previous c~op years and the other
cultivars.
This natural division of the Nep-2 cotyledon
explained its high split score in the processed beans of
that year.
Plant breeders should be aware of that phenomena
because it can not beseen wit~ seed coat intact.
Under the SEM, the cotyledon which is the major part of
the seed reveals parenchymatr~s cells (Figures 21A , A ,
l
2
81' 82' Cl' C2)·
Irregular shipes and arrangement of the
parenchyma cells with reserve materials in form of starch
granules are observed in the three cultivars.
The parenchy-
ma cells length and width ranged from 39 tô 94 ~m and 30 to
56 ~m for Sanilac, 63 to 104 ~m and 31 to 57 ~m for Nep-2
and 46 to 68 ~m and 29 to 42 ~m for San-Fernando, respec-
tively.
San-Fernando shows shorter and'thinner parenchyma
cells.
These cells are more tightly surrounded· by the
middle lamella with the starch granules firmly embedded in
the protein matrix (Figures 21C l , C2). This could imply

---

112
"'.
_
r
1
j
1
{
. INNER
-
. - -
.1
Figure 20.
Scanning electron micrographs of 1980 dry Nep-2
bean cotyledon outer and inner side surfaces
structures (30x); A = cotyledon outer side
surface; B & C = cotyledon inner side surfaces.

---

113
8ANILAC
r ~
.. "
. ..' ........
)-/ '
Figure 21.
Scanning electron micrographs of dry bean seed
cotlydon cross-section structures (1600x).
Al = Sanilac longitudinal cross-section; A2 =
Sanilac transverse cross-section; Bl = Nep-2
1 0 n9 i t ud i na l c r 0 S s - sec t ion; B2 = Nep.,.. 2--t r ans .,.-
verse cross-section; Cl = San-Fernando longi-
tudinal cross-section; C2 = San-Fernando
transverse cross-section; CW = cell wall; ML =
middle lamella; P = protein matrix; S = Starch
granules.

---

114
".
that there may be more parenchyma cells in the San-Fernando
than in the other two cultivars.
The f1rmly packing of the
cel l s ' i s a go 0 d i nd i ca t ion 0 f the fi rm e rte xt ure 0 f th a t
bean cultivar.
This firm packing of cells could cause
increased resistance to shear.
Although Sanilac and Nep-2 have organized parenchyma
cell structures which are amorphously bOund to consecutive
cells, Nep-2 possesses more vascular bundles scattered
throughout the seed cotyledon as se en at lower magnification
(Figure 228 , 8 ).
No topographical difference was observed
1
2
between the longitudinal and transverse section structures
of the individual bean cultivar.
The starch granules:, although irregular in form and
s ha pe, r a n9e d f rom l 2 t 0 28 1-1 m, 3 t 0 39 ~m - and 9 t 0 22 1-1m
in Sanilac, Nep-2 and San-Fernando, respectively.
The
San-Fernando starch granules are of smaller size.
This
/
may have been caused by the firm packing of the parenchyma
1.
cells in the cotyledon during the maturation of the seed,
hence preventing a full expansion of the starch granules in
the process.
The boundaries between the cell wall of adjacent cells
which could not be distinguished in naw lima beans by
Rockland and Jones (1974) are clearly seen here as also
observed in cowpea by Sefa-Dedeh
. ,

---

l l 5
....
SANILAC
NEP-a
Fig ure 22-.
Scanning electron micrographs of dry bean seed
cotyledon cross-section structures at lower
magnification (400x).
Al = Sanilac longitudinal'
section; A2 = Sanilac transverse section~ 81 =
Nep-2 lQngltudinal section; 82 = Nep-2 trans-
verse section; Cl = San-Fernando longitudinal
section; C2 = San-Fernando transverse section;
V = vascular vessels; S = starch granules; C ~­
cell; P = protein matrix.

---

116
Water Absorption on Soaking
Dry beans from the individua1 year (1978 s 1979 s 1980)
samp1es with their respective moi sture content were submitted
to co1d water soaking tests (Table 16).
The water absorption
curve pattern is simi1ar to those reported on black beans
by Quast and da Silva (1977a s b) and cowpea by Sefa-Dedeh
(1978).
Sani1ac and Nep-2 have the highest water absorption
rate compared to San-Fernando (Appendix Figures Al and A )
3
except in the 1979 beans where Nep-2 water absorption rate
was 1ess th an that of San-Fernando (Appendix Figure A2).
There was no exp1anation of this from the examination of
the seeds under SEM.
In genera1 s we cou1d attribute the
effect to weather s maturation and storage conditions of the
seeds.
But we cannot imp1y moisture content of the seed as
a factor in the condition because there was no difference
in water content within that year among the cultivars.
The
difference in the water absorption rate among the three
bean cultivars under investigation cou1d main1y be associa-
ted with the differences found in their structural topo-
graphy.
As discussed ear1ier s the seed coat in San-Fernando
presents a resistance to water penetration because of its
structure.
Despite the initial moisture of the beans
Sani1ac achieved a more rapid uptake of water than the
other two (Figure 23 and Appendix Figures Al s A2 and A3 ).
The fact that beans stored at high moisture content tends
i
J

---

Table 16.
Percent water absorption 1 of dry beans soaked in ambient temperature water
(1:1, tap water:distilled water) for up to 90 minutes by crop years (1978,
1979 and 1980).
1
Sean Cultivar
Sanilac
Nep-2
San-Fernando
Crop Years
Crop Years
Crop Years
Soak time
(min. )
1978
1979
1980
1978
1979
1980
1978
1979
1980
o
8.1
6.5
5.9
8.8
6.5
5.8
9.2
6.7
5.8
1 5
35.4
21.1
23.1
26.0
14.1
22.1
16.9
16.9
12.8
30
47.7
40.1
40.5
41.6
15.6
31.7
21.8
18.2
13.7
.......
.......
45
47.7
40. 1
47.9
44.2
1 5. 7
41. 9
25.9
24.2
15.0
"-J
60
51.4
50.7
51.1
46.5
1 7 .2
46.5
32.2
31.8
1 5 • 1
75
53.1
52.7
52.8
49.8
20.8
49.4
33.8
34. 1
1 6 . 1
90
53.3
53.1
52.8
50.5
21.1
50.5
37.5
40.5
1 7 • 1
n = 3 (3 replicates/soak time/crop year)
1 = The percent water absorption was calculated according to Hosfield, L.G. and Uebersax,
M.A. methods (1980).

---

. ~:'-, .
....
1
70
;
a
IJJ
al
II:: 60
0
en
SANILAO
al
•
et 50
II::
IJJ
!iEP-2
~ 40
~
~ ----"
~ ./ /.~--
SAN-FERHA
~ 30
w
/
_.._-_-0---"-- NOO
--'
--'
0
II::
co
~--"'"
IJJ 20
a.
" .. ..---" - -"'"
'
.
10
"
r
Q
10
20
30
40
50
60
70
80
90
1
J
liME
,
')
.'.'
-- ---r-r-
---------
Figure 23.
Mean percent water absorption pattern of dry bean cultivars (Sanilac, Nep-2
and San-Fernando) soaked in ambient temperature water (1:1, tap water:distilled
water) for up to 90 minutes over three crop years (1978, 1979 and 1980).

---

119
to give a
low
. water uptake on soaking is not applicable
in this study because 1ess than 10% moisture content were
found in the-beans under investigation.
This imp1ies
Sani1ac's easy water absorption through the seed coat and
micropy1e compared to Nep-2 and San-Fernando which possess
a sticky protein membrane on the cotyledon outer surface
preventing the easy translocation of water between the seed
coat and the cotyledon during the hydration process (Figure
158 , Cl)'
The hypothesis presented on cowpeas by Sefa-
1
Dedeh (1978) and soybean by Mayer and Po1jakoff-Mayber
(1975) indicating that highest amount of water absorbed is
re1ated to high protein content is not supported here
because the three cultivars used in this study did not have
any significant difference in their protein content but
there were differences in their water content absorption
rates during soaking.
If protein i5 the chief component
absorbing water in seeds as reported by Mayer and Po1jakoff-
Mayber (1975), the absorption qua1ity of the protein may be
a factor to consider in the hydration of seeds.
The firm
packing of the parenchyma ce11s in San-Fernando bean cou1d
a1so be suggested to de1ay the water absorption through the
entire cotyledon.

---

120
Pasting Properties
Tables 17A, B, and C and Appendix Table lB show the
Brabender Visco/amy1ograph pasting properties of the three
cultivars.
The f10urs from the three bean cultivars (Sani1ac,
Nep-2 and San-Fernando) have Type C pasting curves with no
definite peak which is common1y observed in cerea1s.
This
particu1ar shape is common1y found in 1egumes (Colonna and
Mercier, 1979; Lineback and Ke, 1975; Naviku1 and d'Appo-
lonia, 1979; Suzuki ~~., 1981; Schoch and Maywa1d, 1968;
Vose, 1980).
A11 three cultivars had high initial pasting
0
temperature 72.8 oC, 72.3 C and 77.3 0C for Sani1ac, Nep-2
and San-Fernando who1e f1ours, respective1y (Appendix
Table 18).
The highest value for San-Fernando indicates
the slow swe11ing of the starch granules of this cultivar.
Higher values were a1so obtained with cotyledon f10urs
0
0
o
with 70."6 C, 75.3 C and 75.8 C over the three crop years
for Sani1ac, Nep-2 and San-Fernando, respective1y (Appendix
Table lB).
While Nep-2 initial pasting temperature is
simi1ar to that of Sani1ac in the who1e bean f1our, it is
most simi1ar to that of San-Fernando in the cotyledon f10ur
a1one.
There was no difference in the initial pasting
temperature within the crop years between the who1e bean
and cotyledon f10urs among the individua1 cultivar..
From
the who1e bean f10urs over the three crop years study,
1
1
-~

---

Table 17.
Pasting characteristics of dry bean portions (whole bean and cotyledon) flours
by crop years (1978, 1979 and 1980) and cultivars (Sanilac, Nep-2 and San-
Fernando) in a 400 ml phosphate buffer solution at pH 5.30.
,f
r'-40·'.o;,-- ... -_.~--
.---
--_._-~'-- ~~-------
. .....-. -----
'r
t
--- A'~- -------
t
.i
V1SCOS1TY
1N
BU·
'1
INITIAL
INITIAL
PERPENDICll.AR
AT
AFTER
PEAK
PEAK
PEAK
AT
PEAK AFTER
PASTING
PASTWG
BISING
95"C
15mln.
TIME
DROP
25°C
on
!!Smln.
95°C
cooDRlI of 2~ oC
VARIETIES
nME
TOC.
TIME TOC
PEAK
(min)
(min.)
(min.)
SANILAC
--'
N
whole
--'
32
74.5
85
260
82
385
-
345
260
coatless
31
73
46
95
220 145
375
72
485
455
675
685
640
<,
t
NEP-2
who!e
33
76
43
91
90
180
340
555
555
470,
coatless
r
33
77
44
91
140
\\50
330
68
420
400 715
720
6701
SAN
FERNANDO
whole
3579
- - -
70
225
81
345
-
330
270 '
",,,j
-coatless
34
77.5
44 92
140
\\50
270
74
360
330
600
615
~ ..:;
580 '1
~/;
.- BRABENDER UNITS

---

Table 17.
(cont'd.).

---

---

124
Nep-2 showed the highest viscosity curves and San-Fernando
the lowest with the Sanilac curve intermediate (Figure 24
and Appendix Figures B1A , B
A ).
With the
l
2Al and B3 l
coatless (cotyledon) bean flours, Sanilac demonstrated
higher curves followed by Nep-2 and then San-Fernando
(Figures 24, A2 and Appendix Figures B1A
and
B A ).
l
2 l
except in the 1980 flour samples where Sanilac showed a
lower curve only during the heating period (Appendix Figure
B3A2)~
Considering the individual cultivar, viscosity
curves appear to be higher after removal of the seed coat
in the 1978 and 1979 Sanilac and San-Fernando bean flours
(Figures 25, 27 and Appendix Figures B A
and
~2A2).
1 2
In the year 1980.beans~ this increase did not occur with
Sanilac (Appendix Figure B A ).
Inversely, Nep-2 showed
3 2
a loss in its viscosity strength after ~emoval of the seed
coat (Figures 24A 2, 26 and Appendix Figures B1A
and B A ).
2
2 2
The 1979 Nep-2 beans which presented a low water absorption
in rate pattern (Appendix Figure A ) revealed the highest
2
whole bean flour viscosity of all materials evaluated
(Appendix Figures B,A l , B2Al and B3Al ).
This behavior was
not explainable on the basis of structural topography,
but it could be suggested that the starch granules, unable
to pick up water in the cold stage, absorbed the wat~r to a
large extent during the heating period before they rupture
and
that made the slurry more viscous.
The same behavior
can not be related to starch content because the Nep-2 beans

---

125
TEMPERATURE
(OC)
25
42.5
77.5
9 5 _ 9 5
77.5
'30
42.5
2 5 - 2 5
1000
, At
WHOLE SEAN FLOUR
(f)800
PASTING CURVES
.,,--,N~?-2
~
~----
:lE
. ~600
".,.
/"
--
........
tSAP:I~~<:
••
e.
0:
l&J
....
/ '
..
~400
/ '
,
..
.....
..-
l&J
/
Cl
/
..- ~
...
.......-..
et
....
"
SAN-FERNANDO
~200
~ /
.'
....
.....
(
L.,"
0
15
30
45
60
75
90
105
120
liME (MINJ
TEMPERATURE
rc)
25
42.5
60
77.5
9 5 _ 9 5
77.5
60
42.5
2 5 _ 2 5
1000
A2 BEAN COTYLEDON FLOUR
800
PASTlNG CURVES
SANILAC
Q)
t:
f.
.
;Z 600
NEP-2 ••••;::;".
-- •.
~
~ ....;;/
0:
~./
~400
.... ...,-/
..,:;... '..;..--
:z
l&J
01
--
.'~""
-.
y;.---
~•.Y
SAN-FERNANOO
~ 200
01
1
0
15
45
60
75
90
105
120
TIME (MIN.)
Figure 24.
Mean pasting curves of dry bean portions'
(who1e bean and cotyledon) f10urs by culti-
vars (Sani1ac, Nep-2 and San-Fernando) over
three crop years (1978, 1979 and 1980).

---

SANILAC BEAN FLOUR PASTING CURVES
1
TEMPERATURE (OC)
1000 2,5
4;'5
6?
7J-5
~
9,5
7}.5
6?
4~.5
2;_2,5
(/) 800
!::
Z
:J 600
Cf:
LLI
--'
°400
Z
N
LLI
O'l
CD
.<
Cf: 200
.CD
15
TIME (MIN~
Figure 25.
Mean pasting curves of dry Sanilac bean portions (whole bean and cotyledon)
flours over three crop years (1978, 1979 and 1980).

---

"
NEP-2
BEAN
FLOUR PASTING CURVES
1
TEMPERATURE
(OC)
100025
42.5
60
77.5
95
95
77.5
60
42.5
25_25
,
i
i
,
i
i
i
i
i
•
i
800
WHOLE
(1)
1-
5600
a:
·lJJ
--'
0400
N
:z
.......
LrJ
m
~200
Dl
TIME (MIN.)
..;,
Figure 26.
Mean pasting curves of dry Nep-2 bean portions (whole bean and cotyledon)
flours over three crop years (1978, 1979 and 1980).

---

SAN-FERNANDO BEAN FLOUR PASTING CURVES
TEMPERATURE
(OC)
25
42.5
GO
77.5
95
95
77.5
GO
42.5
2 5 _ 2 5
1000 ,
,
,
•
•
,
,
•
•
,
,
aoo
en
...ZGOO
COTYLEDON
:J
0:::
l ..,,--
/ /
--'
. " . /
N
W 400
. ;
co
o
·z
W
" . "
'"'-
. " ,
- . " , .",
~WHOLE
al
<t 200
a::
r-"
al
o
t5
30
45
GO
75
90
105
120
TIME (MIN.)
Figure 27.
Mean pasting curves of dry San-Fernando bean portions (whole bean and
cotyledon) flours over three crop years (1978, 1979 and 1980),

---

129
of that year did not show any significant difference com-
pared to San-Fernando (Tables 13 and 14).
The retrogradation properties which were observed during
the cooling period of the bean flour slurries showed similar
patterns for the overall study, with San-Fernando always
the lowest curves.
This suggests that San-Fernando would
not influence bread staling if used in bread making, while
Nep-2 and Sanilac bean flours may influence staling due to
higher viscosity curves.
Similar results were reported on
faba bean, lentil and mung bean starches by Navikul and
d'Appolonia (1979).
The continuous setting-back of the
slurry implies a good retrogradation of linear molecules
in the starch granules.
SEM of Soaked Beans
The soaked bean seed coat outer surface and cross-
sectional structure are presented in Figure 28A , A
,
l
2 , Bl
B2 , Cl' C2)·
Sanilac and Nep-2 showed a plastic-like rupture of the
soaked seed coat while that of San-Fernando is a rigid
rupture (Figure 28C l ). The sinking holes in the Nep-2
coat expanded during soaking (Figure 28B ).
The cross-
l
sections show that the palisade cells become amorphous
during soaking (Figures 28A 2 , B2 , C ).
Rupture is observed
2
in the Sanilac coat cross-section.
This implies a II rup ture
through ll the entire thickness of that seed coat during the
soaking process differing from the two isolines seed coat.

---

, 30
BANI LAC
,
NEP-Ii!
BAN-FERNANCO
SAN-FERNANCC
Figure 28.
Scanning electron microscopy of soaked bean seed
coat outer surface (lOOOx) and cross-section
(lOOOx) structures.
Al = Sanilac seed coat
outer surface structure; A2 = Sanilac seed coat
cross-section structure; B, = Nep-2 seed coat
outer surface structure; B2 = Nep-2 seed coat
cross-section structure; C, = San-Fernando
seed coat outer surface structure; C2 = San-
Fernando seed coat cross-section structure; SH =
9inking hole; AP = amorphous palisade.

---

l 31
The soaked bean seeds hilum are a structure is shown in
Figures 29A l , B , and Cl·
l
All three cultivars present ruptured seed coat around
the hilum area and expanded micropyle.
San-Fernando, which
showed a Y-shaped closed micropyle in the dry seed (Figures
16C
and C
l
2 ) reveals an open Y-shaped micropyle in the
soaked seed (Figure 29C 2).
This enlargement of the micro-
pyle contributed to the maximum absorption of water by
San-Fernando beans during the cold and hot soaking prior
to canning.
The micropyle as mentioned in the literature
by different scientists plays an important role in the
water absorption of beans.
In this study that role is well
confirmed and makes a difference between Nep-2 and San-
Fernando compared to Sanilac.
The soaked bean cotyledon structures are shown in
Figures 30Al' A2 , Bl , B2 , Cl and C2 .
Soaked Sanilac struc-
ture shows released starch granules from ruptured parenchyma
cells exposing the middle lamella while soaked Nep-2 and
San-Fernando present intact parenchyma cells although sorne
starch granules were partially released.
More intact
parenchyma cells are present in the San-Fernando cotyledon.
This implies higher potential resistance to shear for San-
Fernando compared to Nep-2 and Sanilac.

---

l 32
SANILAC
NEP-R
SAN-FERNANDO
SAN -FERNANDO
•
Figure 29.
Scanning electron micrographs of soaked bean
seed hilum area stru~ture; Al = Sanilac hilum
area (50x); A2 = Sanilac micropyle (lOOOx);
Bl = Nep-2 hilum area (50x); B2 = Nep-2 micro-
pyle (lOOOx); Cl = San-Fernando hilum area
(50x); C2 = San-Fernando micropyle (lOOOx);
HC = hypocotyl area (50x); H = hilum; M =
micropyle

---

133
"
. . .;. ;'
,': '..~
Figure 30.
Scanning electron micrographs of soaked bean
~
cotyledon structures (400x and 1000x).
Al and
A2 = Sanilac cotyledon; Bl and B2 = Nep-2
cotyledon; Cl and C2 = San-Fernando cotyledon;
C = cell; CW = cell wall; S = starch granules;
ML = middle lamella; l = intercellular space.

---

1 34
SEM of Processed Beans
Processed Sanilac seed coat structure shows a " p l as tic-
like" rupture of the cuticle exposing an outer surface of a
swollen membrane present between the cuticle and the cotyle-
don (Figure 31A, B).
Higher magnification shows a "hill-
1 i ke Il st ru ct ure wh i ch mi 9 ht ha ve be e n cre a te d bye x pan s ion
of the palisade cells during canning.
Similar observations
are seen with the processed Nep-2 seed coat outer surface
structure (Figure 32A, B, C, and 0).
The cuticle of both
cultivars look similar (Figure 31C and 32C) although Sanil c
has a more structured appearance.
The processed San-Fernando seed coat outer surface
shows a rigid rupture of the seed coat with " gr it like"
structure observed over the three crop years (Figure 33A,
B, C and 0).
This is a characteristic difference from the
other isoline, Nep-2, which shows a clear separation of the
cuticle after processing, exposing the outer surface struc-
ture of an intermembrane.
This intermembrane is not the
linea lucida or light line observed between mucilage
stratum and palisade cells by various researchers (Corner,
1951; Hamly, 1932, 1934; Reeve, 1946a, 1946b; Chowdhury and
Buth, 1970) because it is positioned between the cuticle
and the palisade cells.
This could not be seen on the dry
seed coat cross-section.
Processed Nep-2 seed coat outer
surface structure is similar to that of Sanilac.
This

---

1 35
SANILAC
.. ' SANILAC
" - '~'.."
Figure 31.
Scanning e1ectron micrographs of processerl
Sani1ac seed coat outer surface structure; A
and B = cutic1e and intermembrane outer surface
(100x and 400x); C = cutic1e ce11 outer surface
(3000x); 0 = protein intermembrane ce11s outer
surface (3000x); PC = protein intermembrane
ce11s; CU = cutic1e.

---

136
NEP-2
NEP-2
Figure 32.
Scanning electron micrographs of processed Nep-2
seed coat outer surface structures; A and B =
cuticle and intermembrane surface (lOOx and
400x); C = cuticle cell outer surface (3000x);
o = protein intermembrane cells outer surface
(3000x); PC = protein intermembrane cells;
CU = cuticle.

---

137
;1
SAN-FERNANDO
Figure 33.
Scanning e1ectron micrographs of processed
San-Fernando seed coat outer surface structures;
A and B (lOOx and 400x) and C and 0 (3000x)
cutic1ece11 outer surface; CU = cutic1e.

---

l 38
concurs with the observation made from the dry seeds of
these two cultivars.
San-Fernando seed coat maintains its
differential characteristics throughout the processing
system.
This behavior could prevent rapid absorption of
water.
The processed bean cotyledon parenchyma from the three
cultivars show similar irregular shape and size of expanded
ruptured starch granules held within fine protein fila-
ments.
The middle lamella was ruptured during the canning
step, leaving the starch granules free in the cotyledon
(Figures 34A , A ; B , B , Cl and C
l
2
l
2
2 ).
This also has been
observed by other scientists (Bourne, 1976; Hoseney ~ ~.,
1977; Hahn ~~., 1977; Rockland ~ ~., 1977; Rockland
and Jones, 1974).
No visible differences between Sanilac
and Nep-2 were observed following thermal processing.
Processed San-Fernando beans have some of the parenchyma
cell walls still
holding some starch granules together.
This is a great potential of resistance to shear in the
measurement of texture compared to the other cultivars.
Bean Color
The colors of dry and processed beans are presented in
Tables 18A l , A2 , A3 and Appendix Figure lC.
The lightness
of the se e ds s pe ci fie d bY Il LIl val ue sis no t sig nif i ca nt l y
different in Sanilac and Nep-2 through the overall study
(Tables 18A l , A , A
2
3 and Appendix Figure lC).
The two

---

l 39
"-0.
NEP-2
NEP-2
SAN -FERNANDO
.....
(
Figure 34.
Scanning electron micrographs of processed bean
structures; Al and A2 = Sanilac cotyledon (400x
and 2000x); 81 and 82 = Nep-2 cotyledon (400x
and 2000x); Cl
and C2 = SF cotyledon (400x and
2000x); RS = expandea rupture starch granules.

---

Table 18.
Hunter lab color and color differences values (l, al' bl ) of dry and processed
bean cultivars (Sanilac, Nep-2 and San-Fernando) by crop years (1978, 1979 and
1980).
1
BEAN COlOR
Al'
Crop Year 1978
Dry Beans
Processed Beans
Bean
l
al
bl
l
al
bl
a
e
i
k
Sanilac
64.6
_O. 1d
8.7
5a . 19
4.4
14.8
a
j
k
Nep-2
60.2
_a . 1d
11 .4 e
49.7 9
3.8
14.8
b
c
i
1
San-Fernando
14.7
0.3
_0.7 f
17 . 3h
4.7
3.2
~
0
A2 ·
Crop Year 1979
Sanilac
63.0 a
0.3 b
10.9 c
50.7 e
14.5 9
17 . ai
Nep-2
61 .2 a
O. 1b
10.7 c
46.8 e
17.0 g
17 .6 i
San-Fernando
17 .2 b
- O. 1 b
0.3 d
15.5 f
h
-3.0 j
5.0
A3 ·
Crop Year 1980
Sanilac
60.4 a
0.3 c
11 . ad
50.7 f
14.9 h
16. 7i
Nep-2
54.8 a
O.3 c
11. 1d
50.0 f
14.8 h
.17.6 i
San-Fernando
17.8 b
-0.2 c
0.3 e
14. 19
6.6 h
-2.9 j
n = 8 (2 replicates/sample x 2 samples/can x cans/cultivar).
lHunter value l, al and bl '.
like letters denote no significant differences (p~0.05).

---

l 4 l
cultivars have the highest values above 50 (Hunter readings)
while that of San-Fernando is low and below 20 (Tables 18Al ,
A2 , A and Appendix Table lC, and Figure 35 and Appendix
3
Figures Cl' C2 and C3).
The IIlll values are higher in dry
seeds than in processed on es except in the 1978 San-Fernando
beans where the reverse was obtained (Table 18A and Appendix
Figure Cl).
Greenness or redness represented by lI a ll values
is very low in dry seeds but high in processed ones.
Contrary, a reverse process is observed with IIbl! values
which denote the yellowness or blueness (except in 1978 SF
beans) (Figure 35 and Appendix Figure Cl and C2). This implies
that when IIll\\ values decrease the lI al ll values increase
respectively in the seeds during processing.
This is
expected as a result of browning during the thermal proces-
sing as reported by Uebersax and Bedford (1980).
Through
the overall study, dry and processed San-Fernando beans
showed significant differences of IIll1 and IIb ll
l
values when
compared to the other two cultivars (Tables 18Al' A2 , A3
and Appendix Table lC).
Processed Bean Evaluation
Moisture Content
Ouring the canning process water contents reach the
optimum after the hot soaking step in each individual bean
cultivar (Tables 19A, B, C and Figure 36 and Appendix
Table 10 and Figures Dl' O2 and 03). The maximum is

---

oDRY
r
Got
r -
ill PROCESSED
50 ~
i[
f-;;;
.,
"
.
"
,
"..
::l'
.c 40
,'.
"
"
0
"
"
'.'
.'
.
"
:'
a:
:
w 20
--'
1-
Z
r.:
~
r l -
N
;:)
"
"
::I:
~ ~:!
1-:1
,
n>"
, ,
"
, ,
, "
~.
'
"
:...
':
"
:;
0
~l'.'" ~i~:1è:J
SANILAC
NEP-2
SAN-FERNANDO
Figure 35,
Mean Hunter color and color differences values of dry and processed bean
cultivars (Sanilac, Nep-2 and San-Fernando) over three crop years (1978.
1979 and 1980).

---

Table )9.
Qua1ity parameters eva1uation for processed bean cultivars (Sani1ac. Nep-2 and San-Fernando) by crop year (1978.
1979 and 1980).
Qua1ity parameters eva1uation
Legumes
In1t1a1
Wa ter
Wa ter
Drained l
C1ump'
Sp 1it 1
Texture 2
Texture 2
Soak
1
Moisture
Content
Content
Weight
Rate
Rate
Force
Force
Ra t io
~
After
After
9
1 to 3
1 to 5
Height
Kg/1 00 9
Soaking 1
processing 2
(Kramer
~
units)
A.
Crop Yedr 1978
j
Santlac
16.1 a
54.9 b
58.6 c
286.6 d
1. 2e
1f
47.4 h
64.5
1. 9k
a
h
j
k
Nep-2
13.7
53.8 b
68.2 c
284.9 d
.6 e
1f
52.8
71 .7
1. 9
a
c
g
i
SF
15.2
54.3 b
68.3
286.6 d
1. Be
1f
65.3
88.7
1.9 k
.....
B.
Crop Year 1979
..j::::o
w
a
b
c
f
i
m
r
Santlac
17.3
54.7
7l.4
316.8 d
.3
4.5
22.5
30.6 Q
1. 8
a
b
d
h
j
r
Nep-2
15.2
51. 8
69.1 c
316.4
.6
1. 5
30.5 1
41.5 P
1.8
a
b
i
k
r
SF
15.9
52.3
68.4 c
283.3e
1. 89
3.0
48.5
66.0 0
1. 8
C.
Crop Year 1980
d
f
i
k
1
Sanilac
16.5 a
56.2 b
71.7
320.3 e
3.0
4.0 9
25.8
35.0
1:9
a
b
f
g
k
1
Nep-2
17.8
54.6
71.2 d
315.0 e
3.0
4.5
27. Si
37.4
1. 8
d
h
j
m
SF
15.5 a
47.2 c
67. 3
288.ne
2.0 f
3.0 9
53.5
72.8
1.6
ln"· 2 (1 rep1icate/can x 2 cans/cultivars).
2n • 4 (2 rep1icates/can x 2 cans/cu1tivar).
Like 1etters denote no significant differences (p'0.05).
,. .

---

70
~ DRY
60
oSOAKED
...Z
l.lJ
... 50
~ CANNED
Z
o
o
a: 40
lLJ
~;;: 30
--'
...
+=-
z
+=-
UJ
o
20
a:
w
a.
\\0
o '
r·. , ,
r-, ,.,
,-, 1 ."
SANILAC
NEP-2
SAN-FERNANDO
Figure 36.
Percent water content for bean cultivars (Sanilac, Nep-2 and San-Fernando)
in quality parameters evaluation over three crop years (1978, 1979 and 1980).

---

145
only after canning and that is clearly seen in Figure 36 and
Appendix Figures °1 , 02 and 03'
There was no significant
difference between the cultivars for the moisture contents
after soaking (cold and hot) prior to canning and after
canning, except San-Fernando
which showed sorne differences
in the 1980 bean samples.
San-Fernando beans which had a
slow water absorption rate (Figure 23 and Appendix Figures
Al and A3) reached its optimum and maximum in absorption
during the soaking and after canning process respectively
(Figure 36 and Appendix Figures °1 , 02 and °3),
This may
be related to the opening of its micropyle during the
soaking step and also to the heat treatment which caused
rupture of the seed coat and release of the starch granules
from the parenchyma cells as observed under the SEM (Figure
29).
This confirms the role of seed coat (Coe and Martin,
1920; Martin and Watt, 1944; Ott and Ball, 1943; Powrie
~~., 1960) and micropyle (Snyder, 1936; Kyle and Randall,
1964) in the water absorption system of beans.
Although the
soak ratio obtained in this study is lower than those repor-
ted in navy bean by Uebersax and Bedford (1980), there was
no significant differences among the values obtained for
the three cultivars (Table 19A, B, C and Appendix Table 10).
Orained Weight
Orained weight is a function of the equilibrium of
beans and brine in the can as shown in Tables 19A, B, C

---

146
and Appendix Table lD and Figures 37 and Appendix Figure E.
No significant differences were found among Sanilac, Nep-2
and San-Fernando except in the 1979 beans where San-Fernando
differed from the other two with the value of 283.3 9 (Table
19B).
The combined data showed similar results from San-
Fernando beans (Appendix Table lD) which had the lowest
values 286.2 g.
Clump and Split
The 1980 beans scored higher clump and split values
over the three year study.
The split in Nep-2 incurred
dramatically from l to 4.5 score in the 1978 and 1980 beans,
respectively (Table 19A and C).
The difference is explained
by the natural division or splitting of the 1980 Nep-2 beans
observed under the SEM.
The overall score did not show any
significant difference between the cultivars (Appendix Table
l D) •
Texture
The texture evaluated using the Kramer Shear Press
revealed San-Fernando with highest peaks followed by Nep-2
and th en Sanilac (Figures 38 and 39 and Appendix Figures F ,
l
F2 and F3). The peak values converted into force kg/100 9
sample are re~orted in Tables 19A, Band C, Appendix Table
lD and Figure 39 and Appendix Figure F , F
.
The
l
2 and F3
texture of the 1978 and 1980 beans did not differ signifi-
cantly between Sanilac and Nep-2 within those years, but

---

320
-(!)
~
'.11
,
-
.......-
l, 1
"'300
( 1
:I:
(!)
, ,
-
,
w
, "
3:
,"
c
" ,1
~
••
1
o·
~ 280
, 1
•
<t
...0
,(
a:
l ,
. " .
~
, ,
0
0
C
-.....J
1 ,
".•0
,1
0 "
,
1
•
•
260 L
• "
•o •• 1
SANILAC
NEP-2
SAN-FERNANDO
Figure 37.
Drained weight for bean cultivars (Sanilac, Nep-2 and San-Fernando) in
canned quality parameters evaluation over three crop years (1978, 1979 and
1980).

---

100
KRAMER SHEAR PRESS CURVES
- 9 0
t-
'z
« 0
~ 80
Q:-
1JJt-
W
-1
:I: 70
~
~
o
«
<L 60
UJ
0::
55.-..COMPRESSION
a.
~ r\\ CURVATURE
-
50
:I:
lJJ
o 40
«
a::
o
\\
--'
u. :30
+:>0-
l''INFLECTION
C))
a:
POINT
<t 20
UJ
:I:
Cf)
,
.
o
= - '
~.
-
1!6
SANILAC
NEP-2
SAN-FERNANDO
Figure 38.
Mean Kramer-Shear Press curves for bean cultivars (Sani1ac~ Nep-2 and
San-Fernando) over three crop years (1978, 1979 and 1980).
Curve parameters source: Bo11es~ A.O. ~ ~., 1982.

---

gO ..
1-
601-
S.j
....
-
1
.......
•
• •
w
• •
• •
0
• •
• •
~
~
• •
• 1
•
• •
1 1
R%1
ID
• •
• •
• •
CJ
• •
• •
~
• •
• •
• •
• •
~
• •
• •
• •
• •
• •
..~'••'..~...
O·
SANILAC
NEP-2
SAN-fERNANDO
Figure 39.
Texture for bean cultivars (Sanilac, Nep-2 and San-Fernando) in canned bean
quality parameters evaluation over three crop years (1978, 1979 and 1980).

---

150
the 1979 beans showed differences among the three cultivars.
The overall data presented a significant difference among
the cultivars with Sanilac and San-Fernando having the
lowest and highest texture, respectively (Appendix Table 10).
This difference in texture is a main factor in the evaluation
of the seeds under investigation.
The parenchyma cell
packing or arrangement inside the cotyledon seen in Figures
21 and 22 is a prime factor in the process.
Differences
revealed by the SEM microstructural examination showing 1)
firm packing of parenchyma cells in San-Fernando cotyledon
and 2) non-complete liberation of the cells and starch
granules from the middle lamella prior to retort treatment
contributed to the higher texture of that variety when
compared to the other two.
That was evident when the SEM
study on the processed beans revealed sorne starch granules
still being held by the cell walls (Figure 34).
San-Fer.:.
nando's rigid ruptured seed coat after processing could also
contribute to the resistance to shear leading to an increase
in the texture measurement while the " pl as tic-like" seed
coat in Sanilac and Nep-2 would not give such resistance.
These results support the assumptions of Adams and Bedford
(1975) in that at least a portion of the variability observed
among lines depends on genetic differences without discount-
ing that the harvesting and storage conditions, the initial
moisture level of the seed, the length of processing, the
temperature of the soak, the hardness of the water and the

---

l 51
character of the added fluid all play an important role in
that variability.
The study of texture based on single bean cooking time
is given in Table 20.
Results indicated that it takes more
time to penetrate the whole seed than seed without coat.
San-Fernando revealed the highest time required for the
fall-pin to penetrate the seed.
This indicates again the
resistance of its seed coat to intrusion.
There was no
significant difference between Sanilac and Nep-2 cooking
time in this experiment.
Although texture differences among the three cultivars
under investigation are characterized in this study, the
process of resistance to shear in legumes, in general, still
needs further attention.
Plant breeders and food scientists
are well qualified to cooperate in this study.

---

152
Table 20.
Texture based on single bean cooking time
(minutes) for bean cultivars (Sanilac, Nep-2
and San-Fernando.
MEAN COOKING TIME
(minutes)
Seed Coat
Varieties
Seed Co~t Intact
Partially Removed
a
c
Sanilac
43.0 ± 2.4
22.6 ± l .2
c
Nep-2
41.6 ± l . Oa
21 .8 ± 2.6
b
San-Fernando
85.5 ± 6.2
31 .9 ± 3. l d
n = 40 (4 replicates/variety x la seed-cup/cultivar).
Like letters in column denote no significant differences
(p~0.05) among cultivars and within seed portions.

---

CONCLUSIONS
The results reported in this dissertation on the proxi-
mate analysis; microstructure of dry, soaked and processed
beans; texture; and other processing evaluation parameters
of Sanilac, Nep-2 and San-Fernando have provided valuable
information and the following conclusions can be drawn.
Proximate analysis over three years samples revealed
that there were no significant differences among the culti-
vars under investigation with the exception of different
crude fat contents in the 1979 and 1980 beans.
Sugar analyses showed high hexose and stachyose contents
in all the three cultivars of whole beans.
Raffinose and
stachyose were not detected in the seed coat of any cultivar.
This indicates that the color of the seed does not affect
the sugar content in the seed coat.
The hydration study resulted in San-Fernando having the
slowest absorption rate curve while its protein content,
which is the major absorption fraction of a bean, did not
differ significantly from the other two cultivars.
Reduced
hydration of San-Fernando was attributed to seed coat
structural differences.
The scanning electron microscopy study on the dry seeds
of the three cultivars showed highly organized structures
153

---

154
commonly found in legume seeds.
The different anatomical
structures observed could well be used to explain sorne of
the different functional property behaviors such as water
absorption and texture which occur between the two isolines
Nep-2 and San-Fernando.
From these studies, it is concluded
that San-Fernando seed coat outer surface, seed coat pali-
sade:hour-glass thickness ratio, seed coat inner surface and
cotyledon outer surface interlocking, all
have significant
effect on the seed water absorption.
The micropyle and the hypocotyl area can be used as
indices of identification of the three cultivars because of
their specificity for the individual cultivar.
San-Fernando
beans possess characteristic micropyle openings during
soaking which contributes to its water absorption balance
during canning.
The non-ruptured parenchyma cells observed
in the soaked San-Fernando cotyledon are an indication of
increased resistance to shear in that cultivar compared to
Sanilac which possessed complete release of the starch
granules from these cells.
In addition, the rigid seed coat
and the partially intact starch granules in the San-Fernando
processed beans contributed to the increased resistance to
shear (increased firmness).
These distinct observations
could help plant breeders in the choice of their lines of
selection among bean materials for a particular processing
quality characteristic.

---

155
The bean 1ightness of co1or (L value) decreases during
processing whi1e redness (+a
value) and ye110wness (+b
L
L
value) increase
except in San-Fernando beans where there
were fluctuation in values from year to year.
This is an
expected process which takes place under the heat treatment
causing browning.
No significant differences were obtained among the
cultivars for the moisture content after soaking and proces-
sing.
The processed product revea1ed a high drained weight
and low texture for Sani1ac and Nep-2 but the reverse for
San-Fernando.
The texture differences observed under the Kramer Shear
Press between the two iso1ines can be attributed. when using
a scanning e1ectron microscope (SEM). to the structural
differences obtained in their seed coat and parenchyma ce11
arrangement.
Longer time is required to cook San-Fernando
beans according to the single bean texture study.
The viscosity study indicated high initial pasting
temperature with type C curves for a11 the cultivars.
The
remova1 of the seed coat increases the viscosity potentia1
of Sani1ac and San-Fernando contrary to Nep-2 which showed
a decreased viscosity.
Seed contribution to viscosity
varied therefore with cultivar and showed a difference
between the two iso1ines.
This work provides fundamenta1 data regarding genetic.
physico-chemica1 and structural characteristics of se1ected

---

156
dry beans.
These data are of significant value for inducting
both genetic (biological) and technological
(physical)
changes to improve the quality of dry edible beans.
Coopera-
tive research efforts among plant breeders and food scien-
tists could contribute to improved culinary and nutritional
quality of cooked or processed beans and result in reduction
of the energy required to prepare beans for human consump-
tion.

---

RECOMMENDATIONS FOR FUTURE RESEARCH
1.
The work on the proximate composition provided crude
estimation of the bean constituents.
Further work on
the starch components (amylose and amylopectin) ratio
and the protein quality may help in understanding sorne
of the specific functional
properties such as water
absorption of the seeds and wettability of the flour.
2.
The microstructure studies revealed information on the
seed coat, micropyle, cotyledon and parenchyma cells of
the dry, soak and thermally processed seeds.
Work in
this area on the seeds at different storage temperatures
and humidities may provide information on the structural
changes taking place during storage of legume seeds.
3.
The processed bean evaluation indicated major texture
differences using Kramer Shear Press and single bean
texture measurement instruments.
Further study with
Instron or Wedge texture measurement instruments, may
give additional information on the resistance of indi-
vidual:beans to shear forces.
4.
The study of pasting characteristics of the bean flours
showed variable behavior of-the strain Nep-2 from year
to year.
Further work in this area is needed for the
use in new product development to ascertain the effects
157

---

158
of
rop location and season on flour pasting characteristics.
In general, more investigations on the physical charac-
teristics of dry edible bean are needed to provide infor-
mation which could be used by plant breeders to develop
uniformly quick cooking beans for consumers.
The quick
cooking character would be of most benefit to bean consumers
in lesser developed countries where fuel
is scarce and beans
often require a prolonged cooking time to render them
edible.

---

REFERENCES CITED

---

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1972.
Determination
of oligosaccharides in sorne edible legume seeds.
J.
Jpn. Soc. Food Nutr. 25:25.
Thanamunkul, D., Tanaka, M., Chichester, C.O. and Tung,
C.l.
1976.
Degradation of raffinose and stachyose in
soybean milk by a-galactosidase fromMortierella
vinacea.
Entrapment of a-galactosidase with;n poly-
acrylamide gel.
J. Food Sci. 41 :174.
Theravuthi, W., Johnson, C.E. and Von Elbe, J.H.
1974.
A research note: Oifferentiation of perfection, Alsweet
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J. Food Sci. 39:418j
Tobin, G. and Carpenter, K.J.
]978.
The nutritional value
of the dry bean (Phaseolus vulgaris): A literature
review.
Nutr. Abstr. Rev. Sere A 48:919.

---

176
Uebersax, M,A. and Bedford, C.L~
1980.
Nayy bean proces-
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of canned beans.
Research Report 410.
M.S.U. Agr.
Expt. Sta., E. Lansing, Mi.
United Nations Advisory Committee on the Application of
Science and Technology to Development.
1968.
IIInter-
national Action to Avert the Impending Protein Crisis ll •
United Nations, New York.
Valdemiro, C.S., Pedro, L.A. and Luiz, D.A.
1979.
Nutri-
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(Phaseolus vulgaris).
J. Food Sci. 44;1306.
Van Hofsten, A.
1972.
Ultrastructure of protein ri ch rape-
seeds.
J. L11trastr. Res. 38:198.
Varriano-Marston, E.
1981.
Integrating light and electron.
Cereal Foods Horld. 26:558.
Varriano-Marston, E. and Elisa de Omona.1979.
Effects of
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J.
Food Sci. 44:531.
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1971..
Degossypolised cottonseed flour the liquid
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J. Am. Oil Chem. Soc. 48:611.
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1971a.
The Ottawa texture measuring system.
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1971b.
Modernization of texture instrumenta-
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1971.
Sorne measurements of baked bean texture.
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KLA-OCG.
Voisey, P.W. and deMan, J.M.
1976.
Applications of instru-
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1971.
Tex tu r e 0 f ba ked beans -
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J.
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1973.
Measurement of pea
tenderness.
II. A review of methods.
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---

'"
J
177
Von Ardenne, M.
1938.
The scanning electron microscope -
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Z. Tech. Phys. 19:407-416.
Vose, J.R.
1977.
Functional characteristics of an inter-
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Cereal Chem.
54:1141.
Vose, J.R.
1980.
Production and functionality of starches
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Cereal Chem. 57:406.
Walls, E.P.
1936.
Grading and canning tests for raw peas.
Canner. 84:7.
Watson, C.A. and Dikeman, E.
1977.
Structure of rice grain
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Cereal Chem.
54:120-130.
Wolf, W.J.
1970.
Scanning electron microscopy of protein
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J. Am. Oil Chem. Soc. 47:107.
Wolf, W.J. and Baker, F.L.
1972.
Scanning electron micro~
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Cereal Sci, Today, 17:125.
Wursch, P.
1977.
Analyt. Biochem. 77:265.
Wurzburg, O.B.
1968.
Starch in the Food Industry.
In
Furice, T.E. (editor)
Handbook of Food Additives, pp.
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The Chemical Rubber Co., Cleveland, Ohio.
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1965.
The ultrastructure of cotyledonary
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J. Cell
Biol. 25:193.

---

APPENDICES
l'1
- !

---

1
Table lA.
Percent sugars (hexose. sucrose. fnosftol. rafffnose and stachyose) content (dry basfs) of bean cultfvars
(Sanflac. Nep-2 and San-Fernando) fro three crop years (1978. 1979 and 1980) by portfons (whole bean. cotyledon
and seed coat).
Hexose
Sucrose
Inosftol
Ra fff nose
Stachyose
Seed Portfon
Seed Portfon
Seed Portfon
Seed Portfon
Seed Portfon
Whole
Cotyledon
Seed Whole
Cotyledon
Seed Whole
Cotyledon
Seed Who1e
Cotyledon
Seed Whole
Cotyledon
Seed
Coat
Coa t
Coat
Coat
Coa t
San 11 ac
a
a
1.2 a
1. Oa
0.8a
1.9
2.5 a
0.3 a
0.6 a
0.6 a
O.la
O.Sa
O.Sa
NO 1
2.0
2.4 a
NO
a
Nep-2
1. 3a
1. 3a
0.7 a
2.2 a
2.7
0.2 a
0.4 b
0.3 b
O.Oa
0.4 a
0.3 a
NO
2.1 a
2.2 a
NO
.......
SF
a
1.6 a
1.4
0.6 a
2.1 a
2.6 a
0.2 a
O.Sab
0.4 ab
O.Oa
0.4 a
0.4 a
2.0 a
2.1 a
-....l
NO
NO
(Xl
n • 27 (3 replfcates/seed portfon/cultfvar x 3 fnjections/replicate x 3 years)
lND • Nondetectable
Lfke letters in column denote nonsfgnlffcant dlfferences (p)O.OS) among cultfvars and wlthfn seed portions

---

- 1
1
.. ~
.,..
,,~:
60
Q
1&1
".__ ~ _._ _ 1.S~I~__
al 50
a:
""
--~
l-
0
(1)
~NEP-2
/ . /
al
~
. -
ce 40
/
a:
••••••••
1&1
/
............
.•. ...... -,
!ë 30
•• ••• SAN-FERNANDO
..
~
//
.
.......
.... ......
.....,
..•....
1.0
1
!Ë 20
1&1
j.
... .•..
U
a:
1
~ 10
o
10
20
30
40
50
60
70
80
90
1.
TIME (MIN.'
-------_. ----- ~-"
" - - -
Figure Al.
Percent water absorption pattern of dry bean cultivars (Sanilac, Nep-2 and
_____ ._.San-Fernando) soaked_;n ambienttemperature water (1:1, tap water, dist111ed
water) for up to 90 minutes for crop year 1978.

---

70
!
60
{ SANILAC
o
&&1
CD 50
.--
/
.-------------
De
o
/ '
o
al
" /
-c 40
•••
/
.
~
.
a:
/"
_ ••••• - ···..:..·SAN-FERNANDO
11.1
/
!ë 30
.-
......
~
/
..
co
o
/
..•••
..
..
Z 20
•.........•.....
/ -
NEP-2
..
~
.
~
De
~ 10
o
10
20
30
40
50
60
70
80
90
TINE (MIN.)
__ -'-
.J
Percent water absorption pattern for dry bean cultivars (Sanilac. Nep-2 and
Figure A2.
San-Fernando) soaked in ambient temperature water (1 :1, tap water:distilled
water) for up ta 90 minutes for crop year 1979.

---

------ ----- '-- .. - -_._-
---~---- - - - - -------.
----._--_._- -"---
,
70
•,
l,
1
60
Q
1&1
al 50
G:
o
'"
al
< 40
G:
1&1
~ 30
Ir
--'
<Xl
~
--'
20
1&1
Col
.•...........•.......
• . • • • •••.•. • •• • • • •• • • • • • •
~ SAN-FERNANDO
a:
•........ ..•..
.' .
~ 10
10
20
30
40
50
60
70
80
90
TINE (MIN.)
- ------. --
- - - -
Figure A3'
Percent water absorption pattern of dry bean cultivars (Sanilac, Nep-2 and
San-Fernando soaked in ambient temperature water (1;1, tap water:distilled
water) for up to 90 minutes for crop year 1980).
- - - - - -
._--- - - - - - - -
-~
- .._--
-
--.._
-
--..- - - - - - - .. -- ----
-_._---------_.-
_._------
----~

---

182
Table lB.
Pasting characteristics of dry bean portions
(who1e bean and cotyledon) f10urs over three
crop years (1978, 1979, 1980) by cultivars
(Sani1ac, Nep-2 and San-Fernando) in a 400 ml
phosphate buffer solution at pH 5.30.
Initial
VISCOSITY IN (BU)l
L:egumes
Pasting
TOC
at
after
at
After
95 0 C
15 min
25 0C
15 min
at 25 0 C
Sani1ac
Who1e
72.8
118
258
468
410
Cotyledon
70.6
118
331
653
623
Nep-2
Who1e
72.3
190
411
721
667
Cotyledon"
75.3
105
265
623
590
San-Fernando
Who1e
77.3
53
151
328
296
Cotyledon
75.8
93
216
538
513
n = 6 (2 rep1icates/seed portion/cultivar x 3 crop years)
1 = Brabender units

---

TEMPERATURE (OC)
25
41
59
77
9 5 - 95
77
59
41
2 5 - 2 5
1000 r----r---T""""---r--~r---=:::..=r:---;;:....--:;:..--:;;:....--:;:.:=::.:;
Al ,WHOLE BEAN FLOUR
800
PASTlt4ëivCURVES
co
!:600
z
; )
Il:
~400
_----SAN1LAC"
... ....
z
..".,.
..... ... :-:-. -:-- --"
.............. ...
t · .. :-:-.,,=,,-
1&1
CD
~:-:-: • • •••
SAN-FERNANoO"'· .
~200
/....
CD
,/..
/.-.
0 .....--~15:---~30~::::;Si;~:::·:.::-:.:;:~~--~60---7.1..5--
......
9L..0----"10-5----I120
TILtE (MIN.)
TEMPERATURE (OC)
25
41
59
77
9 5 _ 9 5
77
59
41
2 5 - 2 5
1000 r--:---:"1:----r--~:::.::::::..:;:~-:.;---:;:=--.......:;....-.:;:.=~
A2 BEAN COTYLEDON FLOUR
800
. PASTING CURVES
co
!:600
z
; )
Il:
:g400
z
1&1
CD
~200
CD
O....---:15;---::30~·:::::=:';""4~5~-~6":"0---7..l.5---9""0-----"10L.5--...J120
TildE (MIN.)
Pasting curves of dry bean portions (whole
. Fig ure BL".
bean and cotyledon) flour oy cultivars
(Sanilac, Nep-2 and San-Fernando) in a
400 ml phosphate buffer solution at pH
5.30 for crop year 1975~
!~

---

184
TEM~ERATURE (·C)
25
41
59
77
' 9 5 - 9 5
77
59
41
/000 r---r---"T""'---,---r---....,.---r---r----,...~-"T"""-.....,
Al
WHOLE BEAN FLOUR
800
PASTING CURVES
.
..
-- -
--
CI)
!:600
z
:1
Il:
~400
z
11.1
CD
,....
....
=200
ID
........ ••••••• SAN-FERNANDO
......
.....
30
45
60
75
90
105
120
TIME CMIN.)
TEMPERATURE (·C)
59
77
95
95
77
59
41
2 5 _ 2 5
2~5:..__~41~_~
1000r
_
_2;~-~==:.;::--!:/.---=~-...:~-~=~
BEAN COTYLEDON FLOUR
800
PASTING CURVES
SANILAC~
CI)
!:600
""
·z
""-
.,-NEP-2-
:1
"7
. . . . . . .
Il:
/. ~•• ''':'sAN:FERNANoO".
""",,
.......-;..
~400
z
.
.
11.1
","" ""
ID
( /
~200
ID
J
_ / .
O....
--
---:15~-~~:::0~=::::...:4~5----=6:-0--~7~5~--9J..0---IOL5---IJ20
TIME (MiN.)
Figure 82,
Pasting curves for dry bean portions (whole
bean and cotyledon) flours by cultivars
(Sanilac, Nep-2 and San-Fernando) in a 400
ml phosphate buffer solution at pH 5.30 for
crop year 1979.

---

1 85
TEMPERATURE C·C)
2~o5~.....:4~1__;59:-'_~77~_~9~5==:9;5:..-_~7;"7_-=5~9_~4~1_'-':2;5=~25
'000 r-
WHCLE BEAN FLOUR
800
PASTING CURVES
0)
I NEP 2
-
• !:eoo
:z:
: )
JI::
......
.-.----
-_SANILAC
"
. ~400
z
-
......
"
'"
............
..' ';':SAN-FERNANOO •
IIJ
......
..'.
CD
:200
-
......
.
......
..
_......
....
CD
.
."",.
-~
.
O.....- - - - - -.......IIIB:.:...
:.:.;·;,.;.:··;,;.··....L..-·_ _- I -_ _- J .
" - -_ _....._ _- - l
~
ro
~
~
~
~
~
~
TIME CMIN.)
TEMPERATURE (~C)
25
41 .
59
77
9 5 _ 9 5
77
59
41
2 5 _ 2 5
'000
A2 BEAN COTYLEDON FLOUR
800
PASTING CURVES
0)
!:600
SANILAC
:t!:
: )
1 ~---
~...-:.
EP-2
........
l!:
~
~400
... ~
z
••• ·~FERNANOO
IIJ
CD
~200
CD
0
15
30
40
60
75
90
105
120
TUdE (MlN.)
'Figure 8 ,
3
Pasting curves of dry bean portions (whole
bean and cotyledon) flours by cultivars
(Sanilac, Nep-2 and San-Fernando) in 1 400
ml"phosphate-buffer solution at pH 5.30 for
cr 0 p ye a r 1980 '.
\\
,
1
i

---

186
.....
Table le'
Hunter Lab color and color difference values of
dry and processed bean cultivars (Sanilac, Nep-2
and San-Fernando) from three crop years (1978,
1979 and 1980).
Dry Beans
Processed Seans
Sean
L
aL
bL
L
aL
bL
a
c
e
Sanilac
62.7
0.2
0.2
50.5 9
11. l i
16.2 j
a
j
Nep-2
58.7
o. l d
11 . le
48.8 9
11 .9 i
16.7
b
h
j
k
San-Fernando
16.6
O.Od
_0 . l f
15.7
5.2
-2.6
n = 24 (2 replicates/sample x 2 samples/can x 2 cans/
cultivar x crop years)
Like letters denote no si9nificant differences (p~0.05).
amon9 cultivars

---

'.
oDRY
r -
60 1-
r -
rn PROCESSED
i-;-;"
50
t- 1 t
~i
"
'.0
1
e
"
J
"
:".
r
','
C 40
1 ~~"',:.
:~,
.":
30
t-
'"
1
::;-
.....J
"
1
:'.
.:.
','
--'
ex>
20
':';
','
.'
"'oJ
, ,
"
~'"
t
','
;1 ' ffi
1-
:~:,~:
;..
'::
" ,
.'
l';.
.....f:.'.'
"
1
Z
10
",
:',
"
.,
- " \\
.::
.::
.,
."
, ,
','
'"
.:.
- J
..
[l
::
.
"T'"
• ,
'"
','
.....
','
'
.. . . .':
..
~'J r.::1
, '
1]
,','
~
".
;0.
.. ..
.~.
I..••J
o
..".i
1.: 0°1
"
-
-"'!
.......
SANILAC
NEP-2
SAN-FERNANDO
Figure Cl.
Hunter color and color differences values of dry and processed bean cultivars
(Sanilac, Nep-2 and San-Fernando) crop year 1978.

---

oDRY
r-
60
r -
t-
ill PROCESSED
~ 50 ~ Il
}:-.
,.,
' . '
".
t
."
40
'.
1 1:.:
o
Il:},
"
"
"
..J 30 tir
, '
'.
· .
a::
.
20
~::..
.'
00
~
w
·.
::
00
.
\\
'.
..
J-
.'
~~:".o,,
' .
'.
• • • •1
\\ :
'0.
Z
10
'.
"
:.
~.
"
, ,
'.'
'.
. ,
'0'
:=:>
:'.
.0.
"
"
.
'.
:c
'."
, .
::
::,1
..
· .
'
:.
~
LI
0
.'
lJ
SANILAC
NEP-2
SAN-FERNANDO
Figure C2 . Hunter color and color differences values of dry and processed bean cultivars
(Sanilac, Nep-2 and San-Fernando) crop year 1979.

---

oDRY
GO
f-
r-
l1J PROCESSED
r -
1
1::::
..Q 50
.'
.,'.'..'::
1• •
-'
c 40 t
'.
~:
.' .
.'
.',
-l 50 1
"
"
'.'
.:
~ 20
:.-.
r::
r-
--'
W
..
.,.:
:..
co
'
t-
lD
.' .
~
r::;
f:!:.
:~.:.
::,
0°.:
Z
.--1:.:
...
10
...
..
'.
P-.
'"
:-0
"
:J
;.
"
o.
1.-'.
:.:
0 0 .
'.
:..
::
-.
..
:.:
~:
'.'
.'
,,:0.: n:
:::c
_.
"
'.
..
'.-0:
"
"
.:
0 0 :
'.
.' .
m
::.
0
"
l::2J
SANILAC
NEP-2
SAN-FERNANDO
Figure C3. Hunter color and color differences values of dry and processed bean cultivars
(Sanilac, Nep-2 and San-Fernando) crop year 1980.

---

Table 10.
Processed bean evaluat10n - three crop years (1978. 1979 and 1980) bean samp1es.
Qua11ty parameters eva1uat10n
Legumes
Initial
Wa ter
Water
Dra1ned 1
C1umpl
Spl1t 1
TextureZ
TextureZ
Soak
Ho 1sture 1
Content
Con ten t
We1ght
Rate
Rate
Peak
Force
Ra t 10
%
After
After
9
1 to 3
1 to 5
He1ght
Kgll 00 9
Soak1ng 1
Proces s 1ng 2
(Kramer
%
%
un i t s )
-
b
g
San11ac
. 15. Oa
55.3
67. ZC
307.9 d
Z.4 f
3.0
31. gj
43.4 m
1. gn
f
1
n
Nep-2
16.1 a
53.4 b
69.5 c
305.4 d
1. 2
Z.5 g
36.9
50.2 1
1. 8
1D
f
g
Cl
SF
16.6a
b
c
n
51.3
68.0
286.Z e
1. 8
2.5
55.8 h
75.8 k
1. 7
ln = 6 (1 repl1cate/can x 2 cans/cult1vars x 3 crop years).
n = lZ (2 repl1cates/can x 2 cans/cult1var x 3 crop years).
L1ke letters denote no s1gn1f1cant differences (p'0.05).
..

---

~ DRY
70 t
-
oSOAKED
60 1-
IJ~
tÏJl
rt1
~ CANNED
1
ri/II
ri 'JI
!VA
50
1-
z
~ 40
z0
(,)
a::: 30
kJ
\\0
~
~
1- 20
z
LLI
u
a: 10
LLl
Go
o 1
1--1 /'li
t8 l/fl
fil
r (j
SANILAC
NEP-2
SAN-FERNANDO
Figure Dl'
Percent water content of bean cultivars (Sanilac, Nep-2 and San-Fernando) in
quality parameters evaluation for crop year 1978.

---

00 DRY
70 t
~
m:1
-
oSOAKED
GO 1-
IlfJ
rhl
r~J
~ CANNED
1
-l//I
IÏJI
rli
50
t-
z
U!
t- 40
z
0
u
a: 30
--'
U!
1.0
N
ti
3: 20
t-
z
U!
u
a: 10
U!
CL
o
SANILAt
NEP-2
SAN-FERNANDO
Figure D2, Percent water content for bean cultivars (Sani1ac, Nep-2 and San-Fernando) in
dry, soaked and canned beans qua1ity parameters eva1uation for crop year 1979.

---

~ DRY
70
oSOAKED
60
~ CANNED
50
t-
Z
UJ
t- 40
z
o
(,)
a:: 30
......
UJ
\\.0
W
!ri
iJt
t- 20
:z
lU
(,)
a: 10
UJ
A.
o '
ri 'f'
t '
"1
5'
"1
SANILAC
NEP-2
SAN-FERNANDO
".
Figure 03.
Percent water content for bean cultivars (Sanilac, Nep-2 and San-Fernando) in
quality parameters evaluation for crop year 1980.

---

~1,1 SANlLAC
'.0NEP-2
320
rr;11,
..
~
"
SAN-FERN ANDO
fil
,1-
"l-
II
III
I
-
,,'-
,
l,'
l,
2
'1
Il
e 300
-
l-
'l'
1i ~
'1\\
'II
"
'.
Jo-
:z:
'1
'II :.IJ
Cl
\\ 1
\\,
'1 •~.
i:i:i
\\1
:=
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Figure E.
Drained weight for bean cultivars (Sani1ac, Nep-2 and San-Fernando) in canned
bean qua1ity parameters eva1uation by crop years (1978, 1979 and 1980).

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Figure Fl .
Texture for bean cultivars (Sanilac, Nep-2 and San-Fernando) in canned bean
quality parameters evaluation for crop year 1978.

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Figure Fz. Texture for bean cultivars (Sanilac, Nep-Z and San-Fernando) in canned bean
quality parameters evaluation for crop year 1979.
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Figure F ,
Texture for bean cultivars (Sanilac, Nep-2 and San-Fernando) in canned bean
3
quality parameters evaluation for crop year 1980,

---