Plant Foods for Human Nutrition 44: 63-70, 1993.

© 1993 Kluwer Academic Publishers. Printed in the Netherlands.

H y p o c h o l e s t e r o l e m i c effect o f a m a r a n t h seeds (Amaranthus

esculantus)

A. CHATURVEDI, G. SAROJINI & N.L. DEVI

AICRP - Foods & Nutrition, P.G. & Research Centre (Itome Science), A.P. Agricultural
University, Rajendranagar, Hyderabad- 500 030, India

Received 25 October 1991; accepted 20 May 1992

Key words: Amaranth seed (Amaranthus esculantus), Cholesterol, Cholesterol fractions,

Hypercholesterolemia inducing diet (HID), Phospholipids, Total lipids, Triglycerides

Abstract. Hypocholesterolemic effect of amaranth seeds was studied in mate wistar strain
albino rats as against Bengal gram. The results showed that liver weights were significantly
higher in animals with hypercholesterolemia inducing diet. Contrarily, weights of other organs
like brain, kidney, testes and spleen showed higher weights with amaranth and bengal gram
diets. Serum lipid and TG (triglyceride) contents were lower and HDL - cholesterol fraction
was higher and comparable with amaranth seed diet. Feeding with amaranth seed showed a
definite hypocholesterolemic effect.

Introduction
Amaranth greens are considered edible and are consumed by almost every-
body in South India. A number of varieties of curries and snacks are
prepared from these greens. However, the seed is not commonly used in
Southern India, but many people in North, West and Central India make
use of the seed on special occasions. The amaranth seed is a coarse grain;
recent findings have revealed that the seed is a potential source of protein,
iron and dietary fibre. Amaranth protein has been shown to be superior in
quality to other cereals in that it is high in lysine; thus it is well suited for
blending with cereals.
During the last few years, considerable interest has been developed
regarding the roIe of dietary fibre from different sources in reducing the
serum cholesterol in hypercholesterolemic conditions. Studies with the feed-
ing of soluble fibres such as pectin and guar gum have shown significant
decrease in serum cholesterol levels [ 1-4]. On the other hand, fibres of wheat
bran have not shown such results [5-8]. A regulatory role of plant fibre in

Part of work done under All India Co-ordinated Research Programme for the year 1990-199I
ICAR.
64

Table 1. Composition of the diets used for the study with amaranth seeds (in %)

Ingredients (g) Diet 1 Diet 2 Diet 3

HID HID + ASF HID + BGF

Amaranth seed flour (ASF) - 51.0 -

Bengalgram (whole) flour (BGF) - - 59.5
Cornstarch 63.5 26.5 18.0
Hydrogenated fat 15.0 15.0 15.0
Casein 10.0 - -
Cholesterol 1.0 1.0 1.0
Salt mixture 4.0 4.0 4.0
Cellulose powder 4.0 - -
Fat soluble Vit. Mix. 1.0 1.0 1.0
Water soluble Vit. Mix. 1.0 1.0 1.0
Choline chloride 0.5 0.5 0.5
Total 100.0 100.0 100.0

Energy (Kcal) Computed 438 436 430

Protein (g) Estimated 10.2 9.9 10.4

the metabolism of cholesterol by sequestration of bile acids, and thus

lowering the concentration of cholesterol in hypercholesterolemic con-
ditions has been demonstrated [9]. Since the physiological effects of dietary
fibre appear to depend largely on source and composition of fibre, an
attempt was made to study the effect of amaranth seed (Amaranthus
esculantus) flour on serum and tissue lipid profile in hypercholesterolemic
rats.

Material and methods

Amaranth seed (white variety) was purchased from the local market and
cleaned to separate the extraneous matter. It was further washed in water to
remove dust and dirt particles, sun dried for two days and milled into a fine
flour. The proximate composition of the flour was determined using the
standard techniques [10]. The flour was incorporated in the diets of
experimental animals to provide 10 percent protein.

Biological experiments

Twenty four adult male rats ofwistar strain in the weight range of 100-118 g
were used in the experiment. During the experiment, all the rats were housed
in seperate cages and given food and water ad libitum. Initially, all were fed
with a hypercholesterolemia inducing dietary regimen for 40 days (Table 1).
 65

Table 2. Proximate composition of amaranth seed (in %)

Components Estimated value Reported" value

Protein (g) 19.3 15.5
Fat (g) 6.5 7.6
Fibre (g) 9.8b 17.5c
Ash (g) 2.6 3.2
Starch (g) 61.8 64.5
Calcium (g) 0.8 0.2
Iron (mg) 8.2 10.0
Phosphorus (nag) 368.0 455.0
Thiamine (mg) 0.18 0.1
Riboflavin (rag) 0.25 0.21
Niacin (mg) 3.2 1.31
Energy (kcal) 383 476
aRobert Becker (1989).
bIndicates crude fibre.
°Indicates dietary fibre.

After 40 days, six animals were picked randomly, sacrificed and b l o o d (for
serum) was collected from the jugular vein. The viscera o f each rat was
opened and the liver was separated, weighed, a n d frozen for further analysis.
The remaining 18 animals in the three groups (6 rats per group) were
weighed. Animals in each g r o u p were fed with one o f the following diets:

G r o u p 1 - Hypercholesterolemia inducing ( H I D ) diet (Control).

G r o u p 2 - Diet containing cholesterol + h y d r o g e n a t e d fat a n d a m a r a n t h
seed flour (Experimental).
G r o u p 3 - Diet containing cholesterol + h y d r o g e n a t e d fat + bengal
gram (whole) flour (Reference).

A m a r a n t h seed flour and bengal g r a m (whole) flour b o t h served as sources

Table 3. Mean organ weights of rats fed with HID and experimental diets

Diet Period of Body L i v e r Kidney Heart Brain Testes Spleen

feeding weight (g) (g) (g) (g) (g) (g)
(days) (g)
HID 40 185.7 8.3(4.5) - 1.14 - - 0.72
80 249.4 10,2(4.1) 1.49 0.67 1.38 2.29 0.40
HID + Amaranth 80 242.9 8.9(3.7) 1.50 0.76 1.60 2.40 0.49
seed flour
HID + Bengalgrarn 80 244.4 9.6(3.9) 1.53 0.69 1.69 2,55 0.41
flour
HID: Hypercholesterolemia Inducing Diet.
Figures in parenthesis represent liver weight as per cent of body weight.
66
Table 4. F E R and a p p a r e n t digestibility o f H I D and experimental diets

Group Period o f Initial Final Gain in Food FER % apparent

feeding weight weight weight intake digestibility
(days) (g) (g) (g) (g)

HID 40 107.3 187.1 79.8" 446 0.18" -

80 188.7 249.4 60,7 ~ 462 0.14 a 95 ~
HID + Amaranth 40 107.9 170.9 63.0 ~ 439 0.14" -
seed flour 80 168.3 242.9 74.6 ~ 490 0.15 ~ 89 b
H I D + Bengalgram 40 107.9 184.3 78.3" 444 0.18 a -
flour 80 186.4 244.4 58.0 ~ 520 0.1 P 91 ~

HID: Hypercholesterolemia Inducing Diet.

Figures with same superscripts are not significantly different ( p > 0.05).

of fibre and protein in the experimental diets (Table 1). After feeding for
another 40 days, the blood and organs (liver, heart, brain, kidney, testes and
spleen) were collected as mentioned above. The weights of all the organs
were recorded. A gram of liver tissue was preserved in 10% neutral buffered
formalin for histological examination. The remaining liver lobes were
homogenised, filtered and preserved for further analysis.

Lipid analysis

Serum and liver homogenates were analysed for total lipids [11],
triglycerides [12], phospholipids [13], total cholesterol [14] and HDL -
cholesterol [15]. The histopathological study of the liver sections was carried
out by using the standard technique at the Veterinary Biological Research
Institute, Hyderabad, India. Statistical analysis was done using the method
of Snedecor & Cochran [16].

Results and discussion

The proximate composition of the grain (Table 2) indicated a higher level of

protein, fibre, calcium, iron and vitamins than the reported values. The high
dietary fibre content of the amaranth seed was similar to the value reported
for wheat and maize [17].
The mean liver weight of the animals fed with HID diet were higher (Table
3) than were those of the animals fed with the experimental diets. Contrary
to this trend, the weights of heart, kidney, brain, spleen and testes were
higher in the experimental groups. Except for the weights of the hearts, the
weights of all other organs of animals fed with amaranth seed were lower
Table 5. Mean serum lipid components (mg/dl) in rats fed with HID and experimental diets

Group Period of TL PL TG Total-C HDL-C LDL-C* VLDL- HDL-C

feeding C**
(days) Total-C

HID 40 694 a 92,8 a 92.8 a 105.7 a 33.6" 53,5 18.6 0.32

80 715" 72.2 ~ 94.8" 128.4" 20.4 b 92.8 15.2 0.16
HID + Amaranth 80 331 ab 69.28 86.4 a8 62.2 b 32.2 a 11.1 18.9 0.52
seed flour ( - 53.7) ( - 4.2) ( - 8.9) ( - 51.6) ( + 57.8) ( - 88.0) ( + 24.3)
H I D + Bengalgram 80 3488 81.7 ~ 76.l b 70.38 25.0 ~ 28.0 17.3 0.36
flour ( - 51.3) ( + 13.2) ( - 19.7) (-45.2) (+22.5) ( - 69.8) (+13.8)

TL - Total lipids; PL - Phosphoipids; T G - Triglycerides; C - Cholesterol.

*LDL-C = Total C - (TG/5 + HDL-C)
**VLDL-C = Total C - (HDL-C + LDL-C).
Figures with the same superscripts are not significantly different (p > 0.05).
Figures in parentheses represent per cent change with reference to H I D (80 days) value.

O~
Table 6. Mean liver lipid components (mg/g tissue) and liver changes in rats fed with H I D and experimental diets

Group Period of Liver TL PL TG TC PL/TL TG/TC PL/TC Morphological and

feeding weight Histological changes
(days) (g)

HID 40 8.3 a 63.9 a 10.8" 11.3" 12.3" 0.2 1.4 0.9 Slightly enlarged
appearance
HID 80 10.2 b 64.4" 24.3 b 12.8" 9.4 b 0.4 1.4 2.6 Moderate fatty
infiltration, Necrotic
changes
HID + A m a r a n t h 80 8.9b 40.2 b 24.7 b 9.5 b 5.3 ¢ 0.6 1.8 4.7 Normal appearance,
seed flour (-- 37.6) ( + 1.6) ( - 25.8) ( -- 43.6) ( + 50.0) ( + 28.6) (+80.8) No noticeable
changes in liver
architecture
H I D + Bengalgram 80 9.6 b 48.18 24.8 b 9.78 5.5 c 0.5 1.8 4.5 Normal appearance,
flour (-25.3) (+2.1) (-24.2) (-41.5) (+25.0) (+28.6) (+73.1) No noticeable fatty
changes in liver

TL - Total lipids; PL ~- Phospholipids; T G - Triglycerides; TC - Total cholesterol.

Figures with the same superscripts are not significantly different (p > 0.05).
Figures in parentheses represent per cent values with reference to H I D (80 days) value.
 69

than the organ weights of animals fed with bengal gram flour. These dif-
ferences were, however, not significant (p > 0.05).
The weight gain and FER of the animals fed amaranth seed flour were
higher than of the animals fed with HID and bengal gram flour (Table 4).
However, the percent apparent fat digestibility was low, which perhaps was
due to the higher dietary fibre content. Total serum lipid content in the rats
fed amaranth seed flour was significantly lower (333 mg/dl) (p < 0.05) than
bengal gram flour (348 mg/dl) fed rats (Table 5). However, the HID group,
consistantly showed significantly, higher lipid contents (75mg/dl)
(p < 0.05). A similar trend was observed in the phospholipid and total
cholesterol content. Triglyceride level in Amaranth flour fed rats was sig-
nificantly lower (p < 0.05) than the HID group, but higher than the bengal
gram fed group.
Among the cholesterol fractions, rats fed with Amaranth + HID diet
showed an increase in the HDL - cholesterol levels (32.2 mg/dt) which was
even higher than that achieved with the bengal gram diet (25.0mg/dl).
However, in animals on continued feeding with HID diet, there was a
significant (p < 0.05) drop in this fraction (20.4 mg/dl). Concomittantly, an
increase in LDL-C fractions, in HID group and a decrease in amaranth and
bengal gram group was also observed.
A similar trend was also observed in the liver lipid components (Table 6).
In contrast to the HID fed rats, the total cholesterol, PL/TL, TG/TC and
PL/TG ratios also showed a favourable trend in amaranth seed and bengal
gram flour fed animals indicating no abnormality in lipid metabolism. These
observations are in agreement with the morphological and histopathological
changes observed for experimental and HID rats (Table 6). The hypocholes-
terolemic effect can probably be attributed to the preponderance of un-
saturated fatty acids in Amaranth seeds [18].
The study shows that amaranth seed can be used as a hypocholesterolemic
agent. It can be used as grain by itself or in combination with other cereal
grains.

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