Z. Ern~ihrungswlss.

15, 333-339 (1976)

O 1976 Dr. Dietrich Steinkopff Verlag GmbH & Co. KG, Darmstadt
ISSN 0044-264

Nutrition Research Department, National Research Centre, Dokki, Cairo (Egypt)

The use of Lupinus termis L. cultivated in Egypt, as a food

protein supplement
G.N. Gabrial

and S.R. Morcos

With 1 figure and 5 tables

(Received J u n e 3, 1976)

As a r e s u l t of t h e r a p i d i n c r e a s e i n t h e p o p u l a t i o n , t h e d i e t of E g y p t i a n s
suffers f r o m u n s u f f i c i e n t p r o t e i n sources, p a r t i c u l a r l y those of a n i m a l
o r i g i n (1). T h e o n l y s o l u t i o n is to a u g m e n t t h e diet w i t h l e g u m e s which, as
a class, a r e r e l a t i v e l y rich i n p r o t e i n as c o m p a r e d to m o s t o t h e r classes of
stuffs of v e g e t a b l e origin.
A c o n s i d e r a b l e a m o u n t of w o r k has b e e n c a r r i e d o u t on e v a l u a t i n g t h e
n u t r i t i v e v a l u e of some l e g u m i n o u s seeds, a n d a b o u t t h e i r p o t e n t i a l success
as p r o t e i n s u p p l e m e n t i n h u m a n n u t r i t i o n .
L u p i n (Lupinus termis) seeds a r e c o m m o n l y c o n s u m e d i n t h e M i d d l e
East, t h e i r c o n s u m p t i o n is c o n f i r m e d l a r g e l y to t h e p o o r e r class (2). T h e
h i g h p r o t e i n c o n t e n t of t h e seeds m a k e s t h e m a rich s o u r c e of d i e t a r y
p r o t e i n . T h e a v e r a g e a n n u a l crop of l u p i n for t h e y e a r 1973 w a s 4500 tons.
F o r h u m a n c o n s u m p t i o n , t h e r a w b i t t e r seeds a r e b o i l e d i n w a t e r for
h a l f a n h o u r , t h e n s t e e p e d i n r u n n i n g w a t e r for t h r e e d a y s to r e m o v e t h e
b i t t e r t a s t e of t h e seeds. D e b i t t e r e d seeds a r e s l i g h t l y s a l t e d w i t h c o m m o n salt.

Experimental
I. Sampling and anatysis
The r a w lupins used through all these studies were brought from the local
market. Each of the following treatments was carried out on two kilograms
seeds of:
1) Raw l u p i n seeds (R.L.).
2) Raw decorticated seeds (R.D.L.).
3) Boiled decorticated seeds (B.D.L.), in which the seeds were boiled for one
hour before decortication.
4) Debittered decorticated seeds (D.D.L.).
Immediately after respective treatments, all samples were dried in a v a c u u m
oven at 60~ C, moisture, nitrogen, fat, crude fiber, ash and iron content of seeds
were determined according to the method described by A.O.A.C. (4). Calcium
and phosphorus were determined as described by Stu]fins (5).
The amino acids content of samples were assayed using the paper chromatography technique of Levy and Chung (6), after being hydrolysed with
hydrochloric acid according to Khan and Baker (7). Cystine and Methionine
were determined according to the method of Jamalian and Pellet (8). The
colorimetric method of Blauthi, Chaeztnski and Berbec (9) was followed for
the estimation of Tryptophan.
471

334

Z e i t s c h r i / t f/~r Ern~lhrungswissenschaft, Band I5, H e f t 4 (1976)

T a b l e 1. S u p p l e m e n t t o t h e b a s a l d i e t

Diet
No.

A.A. Suppl.

2
3
4
5
6
7
8
9

.
.
.
0,5
0.5
0.5
0.5

Raw

24.7
.

.
.

Raw
decorticated

10.0

.
22.1

.
.

.
.

0.25
0,25
0.25
0.25

Boiled

.
10.0

.
.
.
.
.
22.8
9.25
.
.
.
21.0
9.25
.
.
.
.
.
.
.
.
.
.

.
.
20.6

Debittered

.
.

.
.
10.0

.
21.6

.
.
.
.
.
.
.
19.1
9.25
20.1

10.0

9.25

II. Determination of toxic ]actors

a) Trypsin inhibitors
S e e d s w e r e e x t r a c t e d w i t h a m m o n i u m f o r m a t e - f o r m i c acid b u f f e r a t p H 3.2,
t h e T r y p s i n i n h i b i t o r s w e r e a s s a y e d a c c o r d i n g to t h e m e t h o d d e s c r i b e d b y
L a s k o w s k i (10).
b) H e m a g g l u t i n a t i n g a c t i v i t y o f s e e d e x t r a c t w a s d e t e r m i n e d a c c o r d i n g
to t h e m e t h o d of Leiner a n d Hill (11), u s i n g r a b b i t r e d b l o o d c o r p u s c l e s .
c) T h e p r e s e n c e of a l k a l o i d s w a s t e s t e d u s i n g Wagner's r e a g e n t . T h e i r p e r c e n t a g e w a s d e t e r m i n e d b y t h e m e t h o d of Jenkens, Christian a n d Hanger (12).
d) C y a n o g e n e t i c g l u c o s i d e s w e r e t e s t e d a c c o r d i n g to t h e m e t h o d of B u r n side (13).
e) S a p o n i n e s w e r e d e t e r m i n e d as d e s c r i b e d b y A i n - s h o k a (i4). A l c o h o l i c s e e d
extracts were evaporated under vacuum and the residue was dissolved in normal
saline. A b o u t 1 m l of s o l u t i o n w a s a d d e d to 10 m l of s u s p e n s i o n of r e d b l o o d
c o r p u s c l e s i n n o r m a l s a l i n e (1 i n 40). H e m o l y s i s i n d i c a t e s t h e p r e s e n c e of s a p o n i n s
i n t h e seeds.

III. R a t feeding experiments

F o r t h e d e t e r m i n a t i o n of n e t p r o t e i n u t i l i z a t i o n , t h e m e t h o d g i v e n b y Mil~er
a n d Bender (15), w a s f o l l o w e d u s i n g h o o d e d r a t s w h i c h w e r e c a g e d i n d i v i d u a l l y .
O n e g r o u p of r a t s w a s f e d t h e b a s a l p r o t e i n - f r e e d i e t a s s h o w n i n t a b l e 1, i n
which the protein sources under test were incorporated into the basal proteinf r e e d i e t a t t h e e x p e n s e of m a i z e s t a r c h a t t h e l e v e l to c o n t r i b u t e 10 % c r u d e
p r o t e i n to t h e diet. A f t e r t e n d a y s t h e r a t s w e r e k i l l e d a n d t h e n e t p r o t e i n
u t i l i z a t i o n w a s c a l c u l a t e d a f t e r t h e a n a l y s i s of c a r c a s s e s n i t r o g e n .

Results and discussion

Proximate analysis
Table 2 shows the nutrient content of raw lupin seeds. The crude prot e i n c o n t e n t is 40.36 ~ w h i c h is h i g h a s c o m p a r e d w i t h o t h e r l e g u m i n o u s
s e e d s . S i m i l a r v a l u e s w e r e g i v e n b y T a n n o u s e t al. (16). T h e f i b e r c o n t e n t
of t e s t e d l u p i n w a s f o u n d t o b e 10.8 ~ w h i c h is f a i r l y h i g h . G l a d s t o n e (17)
showed that the removal of the seed coat greatly reduces the fiber content
in lupin.

Gabrial and Morcos, The use o] Lupinu$ termis L. cuttivated in Egypt 335
Table 2. Proximate analysis of lupin seed calculated on dry weight basis*)
Item

g/100 g

Protein (N x 6.25)
Fat
Carbohydrate b y difference
Fiber
Ash

40.36
9.71
27.13
10.78
3.39
rag/100 g
290
356
6.5

Calcium
Phosphorus
Iron
*) Moisture content is 8.63~/o

A c i d h y d r o l y s i s of r a w a n d t r e a t e d l u p i n seeds r e v e a l e d t h e p r e s e n c e
of t h e f o l l o w i n g a m i n o acids, A l a n i n e , A r g i n i n e , A s p a r t i c acid, Cystine,
M e t h i o n i n e , G l u t a m i c acid, G l y c i n e , H i s t i d i n e , T y r o s i n e , L e u c i n e + Isoleucine, Lysine, P h e n y l a l a n i n e , Serine, Threonine, T r y p t o p h a n and Valine
as s h o w n i n t a b l e 3. G l u t a m i c acid, G l y c i n e a n d S e r i n e a r e p r e s e n t i n
a m o u n t s s i m i l a r to those p r e v i o u s l y r e p o r t e d b y J a m a l i a n a n d P e l l e t (8),
a n d b y T a n n o u s et al. (16), w h i l e a s p a r t i c acid is p r e s e n t i n h i g h e r a m o u n t s .
C o m p a r i n g t h e a m i n o acid c o n t e n t of w h o l e r a w seeds w i t h t h a t of t h e
r a w d e c o r t i c a t e d l u p i n i n d i c a t e d t h a t t h e a m i n o acids a r e n e a r l y c o n c e n t r a t e d i n the d i c o t y l e d o n of t h e seed. B o i l i n g t h e seeds r e s u l t e d i n
s m a l l loss for m o s t of t h e a m i n o acids s t u d i e d w i t h t h e e x c e p t i o n of
Lysine and Tryptophan, which exhibited slight increase after boiling.
Table 3. Amino acids content of raw a n d treated lupin as compared with the provisional amino acid pattern recommended by the FAO (1955)*)
Item
Alanine
Arginine
Aspartic acid
Cystine
Methionino
Glutamic acid
Glycine
Histidino
Tyrosino
Leucine -}- Isoleucine
Lysine
Phenyl alanine
Serine
Threonine
Tryptophan
Valine

Raw
175
762
959
45
112
1209
291
90
290
380
215
305
345
166
99
152

*) Calculated as mg/gtotal nitrogen

Deeorticared
188
799
975
49
125
1229
311
102
297
385
225
312
356
180
102
164

Boiled
183
778
970
46
114
1217
289
95
287
380
228
307
352
174
118
160

Debittered FAO
176
770
942
43
112
1194
281
94
284
374
219
302
349
169
107
155

126
144
180
576
270
180
180
90
270

336

ZeitschriSt ]fir Erni~hrungswissenscha~t, Band 15, Heft 4 (1976)

Bandemer and Evans (18) reported that heating kidney bean caused
small losses for most of its amino acids. Myklestadt, Bjoernstadt and
Lei] (19) showed also that most of the amino acid levels decreased as
t e m p e r a t u r e increased. Taira Harue (20) observed during the preperation
of soybean-based foods that Lysine, Cystine, Arginine, T r y p t o p h a n and
Serine were partially lost during heating. The author stated that heating
at high t e m p e r a t u r e for a long time was the most destructive especially
for Lysine and Cystine and that heating with w a t e r decreased the loss
of Lysine and Arginine but not that of the other amino acids. Boiling the
lupin seeds followed soaking them in w a t e r resulted in slight losses for
most of the amino acids with the exception of T r y p t o p h a n which increased
slightly.

Toxic ~actors
Trypsin inhibitors were present in r a w lupin seeds as shown in table 4.
The level of Trypsin inhibitors was v e r y low (96 units/g sample) as
compared with those of other legumes such as kidney bean which contained about 209,000 units/g (21}. Trypsin inhibitors were destroyed on
boiling lupin seeds for one hour. The levels of Trypsin inhibitors in the
r a w decorticated seeds were less than those of the whole ones, which
means that they are mainly concentrated in the dicotyledon of the seed.
Testing the presence of hemagglutinins, cyanogenetic glucosides and
Saponins revealed the absence of these three factors from r a w and treated
seeds. The presence of Alkaloids was detected in the raw seeds (1.32 ~
These alkaloids are localised in the dicotyledones. Boiling the seeds for
one hour did not eliminate their Alkaloids completely. Boiled seeds
contained 1.16 g Alkaloids per 100 g. Debittering the seeds led to the
removal of all its Alkaloids.

Feeding experiments
Table 5 shows the net protein utilization of raw and treated seeds. The
weight curves of rats fed the different diets of lupin are shown in figure 1.
The whole r a w seeds had the lowest N.P.U. values, 42. Decorticating or
boiling the r a w seeds caused no change in the weight of rats and N. P. U.
values. Debittering the seeds improved the N.P.U. values from 42 to 56.
This i m p r o v e m e n t m a y be due to the toxicity of the large amounts of
Alkaloids found in the raw seeds which were eliminated by debittering
the seeds as mentioned before. Two factors m a y be the cause of the low
N. P.U. values of raw, boiled and debittered seeds. These two factors are,
Table 4. Toxic substances present in the raw and treated lupin
Treatment

R.L.
R. D.L.

B. D . L .
D. D.L.

Saponin
test

Trypsin
inhibitor
U/g

Haemagglutinin
ttU/mg

Alkaloids
g/100

Cyanogenetic
glucoside

1.32
1.40

--re
--vo

1.16
0.00

--ve
--ve

--vo

96

--re

--re
--ve
--ve

101
00
00

--ve
--vo
--ve

Gabr~al and Moreos, The use o~ Lupinus termis L. cultivated in Egypt 337
80
I. Non protein
2. R,L.

~. R,D.L

1 9

~. B.D.L
70

.~ 60'

g 50.
.~.
gO"
E RL,

2 R.D.L.
8. B,D.L. A.A.
9 I~QL. 9 A.A.

30
2

g
6
8
Days on diet

10

Fig. 1. See text.

t h e l o w d i g e s t i b i l i t y c o e f f i c i e n t of t h e p r o t e i n a n d t h e i r l o w c o n t e n t of
e s s e n t i a l a m i n o acids, L y s i n e a n d s u l f u r - c o n t a i n i n g a m i n o acids. T h e f o l l o w i n g t a b l e s h o w s t h e l e v e l of L y s i n e a n d s u l f u r - c o n t a i n i n g a m i n o a c i d s
in t h e l u p i n s e e d s as c o m p a r e d to t h e F A D (22) p r o v i s i o n a l a m i n o - a c i d
p a t t e r n e x p r e s s e d in m g / g n i t r o g e n .
Amino acid
Lupin
FAD pattern
Lysine
215
270
Methionine + Cystine
157
270
Table 5. Net protein utilization of raw, treated, and supplemented lupins
Treatment

N P U value

R.L.

42
45
44
56
62
60
63
70

R. D . L .
B. D . L .
D. D . L .
R . L . -~ ; methionine -k lysine
~ . D. L. q- me~hionine q- lysine
B. D. L. q- methionine -b lysine
D. D. L. q- methionine q - l y s i n e

-----raw lupin
R. ]D. L. = raw deeor~ieated lupin
B. I). L. = boiled deoortie~ted
D. D. L. = debittered deeortieated seeds
R.L.

338

Zeitschri~ft fi~r Ern~hrungswissenschaft, Band 15, Heft 4 (I976)

S u p p l e m e n t i n g t h e r a w , r a w d e c o r t i c a t e d a n d b o i l e d seeds w i t h the two

l i m i t i n g amino acids L y s i n e a n d M e t h i o n i n e r a i s e d the N. P. U. v a l u e s up to
62, 60, a n d 63, r e s p e c t i v e l y . Also s u p p l e m e n t i n g the d e b i t t e r e d seeds w i t h
those a m i n o acids r a i s e d the N . P . U . v a l u e to 70, which is m o r e or less
s i m i l a r to t h a t casein. S i m i l a r r e s u l t s w e r e achieved b y Tannous a n d his
g r o u p (16). T h e y s h o w e d t h a t g r o w t h of r a t s fed u n t r e a t e d l u p i n seeds was
i m p r o v e d by r e m o v i n g the a l k a l o i d f r a c t i o n f r o m the seeds or b y i n c r e a s i n g
the l e v e l of d i e t a r y p r o t e i n a n d b y s u p p l e m e n t i n g t h e diets w i t h its
l i m i t i n g amino acids. T h e y a s c r i b e d t h e poor g r o w t h o b s e r v e d in rats fed
u n t r e a t e d l u p i n diet to be due to t h e p r e s e n c e of both a l k a l o i d s and the
poor q u a l i t y of d i e t a r y proteins.
T a k i n g in c o n s i d e r a t i o n t h a t Lupinus termis is g r o w n in poor a n d
r o u g h l y c u l t i v a t e d l a n d s (23), its use in h u m a n d i e t a r y of E g y p t i a n s will
be a good solution for p r o t e i n shortage.

Summary
1. General analysis of the seeds for protein, fats, carbohydrates, fiber and
ash contents were carried out and the results were given in g/100 g dry seeds.
Lupinus termis contained a higher percentage of protein (40.36), 290 rng calcium
and 6.5 mg iron.
2. All the essential amino acids are present in fair amounts with the exception of sulphur-amino acids and Lysine.
3. Cooking the seeds resulted in a slight decrease of most of the amino acids
with the exception of Tryptophan and Lysine, which exhibited a slight increase
after cooking.
4. Of the toxic substances present in the seeds is Trypsin inhibitor, which
was destroyed by heat treatment, and alkaloids, which were removed by soaking
the seeds three days in running tap water.
5. Supplementing the debittered seeds with Lysine and Methionine resulted
in an increase in its nutritive value.

Zusammenfassung
I. Lupinensamen w u r d e auf seinen Gehalt an Proteinen, Fetten, I<ohlenhydraten, Faser- und Aschestoffen analysiert. Die Ergebnisse wurden in g/100 g
f(ir trockenen S a m e n angegeben. Lup~nus termis enthielt einen h6heren Prozentsatz an Protein (40,36),290 m g Calcium und 6,5 m g Eisen.
2. A]le essentiellen Amlnosfiuren mlt A u s n a h m e yon schwefelhaltigen Aminos~uren und Lysin sind in angemessener H O h e enthalten.
3. Das Kochen der S a m e n bewirkte eine leichte Verminderung der Aminosfiuren mit A u s n a h m e yon Tryptophan und Lysin, w o nae_h d a m Kochen ein
leichter Anstieg zu verzeichnen war.
4. A n toxischen Stoffen sind in den S a m e n der Trypsin-lnhibitor, der bet
l-litzebehandlung zerst6rt wird, und Alkaloide enthalten. Letztere wurden durch
dreitfigiges Einweichen der S a m e n unter laufendem Wasser eliminiert.
5. Der N~ihrwert der entbitterten S a m e n wurde durch Zusfitze yon Lysin und
Methionin erh6ht.

References
1. Morcos, S. R., J. Egypt. Pub. Health 41, 15 (1966). - 2. Aykroid, W. R., J.
Doughty, In: Legumes in H u m a n Nutrition, F A O , Nutr. stud. No. 10 (1909). 3. The food balance sheet, Egyptian Ministry of Agriculture, Dept. Statistics
(1972). - 4. A.O.A.C., Official methods of analysis of official analytical chemists

Gabrial and Morcos, The use o~ Lupinus termis L. cultivated in Egypt

339

( W a s h i n g t o n 1970). - 5. Stuffins, G. B., A n a l y s t 92, 107 (1967). - 6. Levy, A. L..

0. Chung, A n a l y t . chem. 25, 396 (1953). - 7. Khan, N. A., B. E. Baker, J. Agric.
Fd. chem. 3, 653 (1955). - 8. Jarnalian, J., P. L. Pellet, J. Sci. Fd. Agric. 19, 379
(1968). - 9. Blautli, J. 0., M. Charazinski, H. Berbec, A n a l y t . B i o c h e m . 6, 69
(1963). - 10. Laskowski, A., M. Laskowski Jr., A d v . P r o t . C h e m . 9, 203 (1959). - 11.
Liener, I. E., E. G. Hilt, J. N u t r . 49, 609 (1953). - 12. Jenkins, Christian, Hanger,
Q u a n t i t a t i v e P h a r m a c e u t i c a l C h e m i s t r y , p. 315 ( N e w Y o r k - L o n d o n 1956). - 13.
Burnside, J. E., Net. Med. 49, 136 (1954). - 14. Ain Shoka, A. A., M. Sc. Thesis,
F a c u l t y of P h a r m a c y , C a i r o U n i v e r s i t y (1971). - 15. Miller, D. S., A. E. Bender,
Brit. J. N u t r . 9, 382 (1955). - 16. Tannous, R. I., S. Shadarevian, J. W. Cowan, J.
N u t r . 94, 162 (1968). - 17. Gladstone, J. S., F i e l d C r o p A b s t r a c t s 23, 123 (1970). 18. Bandemer, S. L., R. J. Evans, J. Agric. F o o d c h e m i s t . 11, 134 (1963). - 19.
Myklestad, 0., J. N. Bjoernstad, R. Leif, Ski, Ser. T e k n o l . U n d e r s , F i s k e r i d i r
( N o r w a y ) 5, 15 (1972). - 20. Taira Harue, J a p . A g r i c . Res. Q u a r t . 7, 267 (19733). 21. Gabrial, G. N., L. Hussein, S. R. Morcos, Qual. P l a n t . P1. F d s . H u m . N u t r .
24, 61 (1974). - 22. F A O , P r o t e i n r e q u i r e m e n t s , R e p o r t of t h e F A O c o m m i t t e e ,
R o m e , Italy, 24-31 O c t o b e r (1955). - 23. Ekardt, W., R. Willi, W. Feldbeim, Z.
L e b e n s m . - U n t e r s . - F o r s c h g . 155, 92 (1974).
Authors' address:

G. N. Gabrial a n d S. R. Morcos
N u t r i t i o n R e s e a r c h D e p a r t m e n t , N a t i o n a l R e s e a r c h C e n t r e , Dokki, C a i r o (Egypt)