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www.wjpps.com Vol 4, Issue 05, 2015.
669
Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
NUTRITIONAL QUALITY OF RAW AND SOAKING COMPARED TO
PRESSURE-COOKED (CICER ARIETINUM L.) SEEDS
K. Sindhu and S. Sumathi*
Department of Biochemistry, DGGA College (W), Mayiladuthurai-609 001,
Tamilnadu, India.
ABSTRACT
Food is the basic necessity of man. If is a mixture of different nutrients
such as carbohydrate, protein, vitamins and minerals. They nutrients
are essential for growth, development and maintenance of good health
throughout life. Chickpea is an important pulse crop with a wide range
of potential nutritional benefits because of its chemical composition.
Raw and processed (soaked and pressure-cooked) seeds of chickpea
were analyzed for nutritional qualities such as carbohydrates, protein,
iron, calcium, vitamin-B, vitamin-C, vitamin-D, vitamin-E, ash and
moisture. A significant difference was seen between the proximate
composition of raw and processed seeds. Based on the results pressure-
cooked method is recommended for chickpea preparation, not only for
improving nutritional quality, but also for reducing cooking time.
KEY WORDS: phytochemical compounds, proximate composition, Cicer arietinum (L.)
INTRODUCTION
Food is a more basic need of man than shelter and clothing. It provides adequately for the
body’s growth, maintenance, repair and reproduction. Food furnishes the body with the
energy required for all human activities-it provides materials required for the building and
renewal of body tissues and the substances that act to regulate body processes. Man’s basic
drive is for food to satisfy his hunger. Food is intimately woven into the physical, economic,
psychological, intellectual and social life of man.
The man of yonder years was unaware of many of the most prevalent diseases of today. The
humanity suffers to the core from the bonds of these novel diseases. Our food habit is the soul
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VVoolluummee 44,, IIssssuuee 0055,, 666699--668833.. RReesseeaarrcchh AArrttiiccllee IISSSSNN 2278 – 4357
Article Received on
27 Feb 2015,
Revised on 21 March 2015,
Accepted on 11 April 2015
*Correspondence for
Author
S. Sumathi
Department of
Biochemistry, DGGA
college (W),
Mayiladuthurai-609 001,
Tamilnadu, India.
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Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
significant reason for this plight. Our ancestors lived completely in harmony with nature with
clean air and water, unadulterated and nutritious food. Of late, we consume lot of polished
rice, fast food and preserved foods. They confuse our biological clock and subsequently all
the physiological processes are disrupted. Our forefathers had never consumed food for taste
but for appetite. They added taste with nutrition in their diets. They included cereals, millets,
pulses, fresh fruits and vegetables. Each item had its own characteristic special feature adding
good value to the food. If any one of the components is consumed in excess then it turns out
into poison. But if taken in appropriate proportion, then it serves as a well balanced, nutritive
and healthy meal.[1]
The word Cicer is a derivative of Greek word Krios referring to the well known Roman
family name Cicero. Arientinum is also derived from the Latin aries which means ram
referring to the shape of the seed, which resembles a ram’s head. The other major chickpea
producing countries include Pakistan, Turkey, Australia, Myanmar, Ethiopia, Iran, Mexico,
Canada and USA. It is found in tropical condition. It needs warm and moist climatic
conditions to propagate.[2]
Chickpea seed is processed and cooked in a variety of forms depending upon traditional
practices and taste preferences. Different domestic processing methods (decortications,
soaking, sprouting, fermentation, boiling, roasting, parching, frying and steaming) was used
to obtain a suitable texture for the consumer, improvement in the nutritional factors, increase
the protein digestibility, increases the bioavailability of nutrients, by inactivating
antinutritional factors, growth inhibitors and haemagglutinin. Cooking brings about a
number of changes in physical characteristics and chemical compositions of dry legumes,
which are commonly cooked by a boiling process before use. Pressure boiling and steaming
can also be used. Prior to cooking, soaking is a preliminary step, it helps soften texture and
shorten the cooking time. High pressure processing technology may provide high quality of
food products flavor, color and biological active components.[3]
The current study was therefore conducted with aim to analyze and compare the nutritional
qualities of raw and soaking chickpea compared to pressure-cooked chickpea.
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Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
MATERIALS AND METHODS
Collection of seeds
Cicer arietinum (L.) Brown seeds were collected from local market. Soon after the collection,
the immature and damaged seeds were removed and the mature seeds were dried in the sun-
light for 24 h and stored in plastic containers in a refrigerator (5°C) until further use.
Sample preparation
The whole seeds of Cicer arietinum (L.) randomly separated into 3 batches (25g each).
First batch (Raw sample)
The seeds were prepared as raw seed (milled on 30 mesh size).
Second batch (soaking sample)
The seed was soaked in distilled water for 6 hours at room temperature and the seeds were
taken at a water ratio of 1:10 (w/v). The seeds are dried ground into uniform fine powder
using milling machine.
Third batch (Pressure–cooked sample)
The seed was soaked in distilled water for 6hours at room temperature and the seeds was
taken at a water ratio of 1:10 (w/v) in a pressure-cooked for 15 minutes. Pressure-cooked
seeds were freeze-dried, milled and stored as a seeds sample. All the powdered sample was
used for phytochemical and proximate analysis.
Phytochemical analysis
Phytochemical analysis were carried out using standard procedures of Harborne (1973) and
Sofowara (1993).[4,5]
Detection of Alkaloids
Mayer’s Test
To a 1 ml of filtrate, few drops of Mayer’s reagent are added by the side of the test tube. The
white or creamy precipitate indicated the test as positive.
Detection of Carbohydrates
Benedict’s Test: To 0.5 ml of extract, 0.5 ml of Benedict’s reagent was added. The mixture
is heated on a boiling water bath for 2minutes and the result was observed. A red precipitates
indicated the presence of sugar.
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Detection of Glycosides
Legals Test
Chloroform (3ml) and ammonia solution (10%) was added to 2ml seed extract. Formation of
pink color indicated the presence of glycosides.
Detection of Protein
Biuret Test
An aliquot of 2 ml of filtrate was treated with drop of 2% copper sulphate solution. To this, 1
ml of ethanol (95%) was added, followed by excess of potassium hydroxide pellets. The pink
color in ethanol layer indicated the presence of protein.
Detection of Amino Acids
Ninhydrin Test
Two drops of ninhydrin solution (5 mg of ninhydrin in 200 ml of acetone) are added to 2 ml
of aqueous filtrate. The color change was observed. A characteristic purple color indicated
the presence of amino acids.
Detection of Phytosterols
Liberdmann-Burchard’s Test
The extract (5 mg) was dissolved in 2 ml acetic anhydride and one or two drops of
concentrated sulphuric acid was added slowly along the sides of the test tube. The formation
of blue green color indicated the presence of triterpinoides and phytosteroids.
Detection of Tannins
Ferric Chloride Test
The extract (5 mg) was dissolved in 5 ml of distilled water and few drops of neutral 5% ferric
chloride solution was added. The formation of blue green color indicated the presence of
tannins.
Detection of Phenols
Lead Acetate Test
The extract (5 mg) was dissolved in distilled water and 3 ml of 10% lead acetate solution was
added. A bulky white precipitates indicated the presence of phenols.
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Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
Detection of Flavonoids
An aqueous solution of the extract was treated with ammonium hydroxide solution. The
yellow fluorescence indicated the presence of flavanoids.
Detection of Coumarins
10% NaOH (1ml) was added to 1ml of the seed extracts formation of yellow color indicated
the presence of coumarines.
Detection of Saponins
Distilled water 2ml was added of each seed extracts and shaken in a graduated cylinder for 15
minutes lengthwise. Formation of 1cm foam indicates the presence of saponins.
Detection of Quinine
Concentrated sulphuric acid (1ml) was added to 1ml of each of the seed extract. Formation of
red color indicated the presence of quinones.
Detection of Cardiac Glycosides
Glacial acetic acid (2ml) and few drops of 5% ferric chloride were added to 0.5% of the
extract. This was underlayered with 1ml of concentrated sulphuric acid. Formation of brown
ring at the interface indicated the presence of cardiac glycosides.
Detection of Terpenoids
Chloroform (2ml) and concentrated sulphuric acid was added carefully to 0.5 ml of extract.
Formation of red browncolor at the interface indicated the presence of terpenoids.
Detection of Phlobatannins
Few drops of 10% ammonia solution were added to 0.5 ml of extract. Appearance of pink
color precipitates indicated the presence of phlobatannins.
Detection of Anthraquinones
Few drops of 2% Hcl were added to 0.5 ml of extract. Appearance of red color precipitate
indicated the presence of anthraquinones.
Detection of Steroids and Phytosteroids
To 0.5 ml of the seed extract equal volume of chloroform was added and subjected with few
drops of concentrated sulphuric acid. Appearance of brown ring indicates the presence of
steroids and appearance of bluish brown ring indicated the presence of phytosteroids.
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Proximate analysis
Estimation of Carbohydrate[6]
0.2-1.0ml of standard solution were pipetted out into different test tube. 0.5ml of sample was
taken in different test tube. The volume was made upto 1ml with distilled water and 4ml of
anthrone reagent was added to all the test tube by dipping them in an ice bath after cooling.
Test tube was shaken and heated in a boiling water bath for 20 minutes. The green colour
formed was read at 640nm.
Estimation of Protein[7]
0.2-1.0 ml of the standard protein solution was pipetted out into different test tubes. The
volumes were made upto 1ml with distilled water. To this 4.5ml of alkaline copper reagent
was added and mixed well kept under room temperature for 10minutes. Followed by 0.5ml of
follin’s ciocalteau reagent 0.1ml of suitably diluted. Copper reagent and allowed to stand for
10 minutes at room temperature 0.5ml of follin’s ciocalteau reagent was added. The blue
color developed was read 640nm after 20 minutes.
Estimation of Iron[8]
0.2-1.0ml of standard solution was pipette out into different test tubes. 0.5ml of sample was
taken in different test tubes. The volumes was made up to 1ml with distilled water 1ml of
each of sodium sulphite and 2,2'dipyridyl reagents was added to all the test tubes. The colour
was formed was read at 520nm.
Estimation of Calcium[9]
Standardisation of potassium permanganate 5ml of standard oxalic acid and an equal volume
of dilute sulphuric acid is pipetted out into a clean conical flask. The solution is heated to
bearable warmth the hot solution is titrated against the permanganate in the burette. Till the
end point is the permanent pale pink colour titration are repeated for concordant values. To
2ml of extract and 2ml of distilled water is pipetted in to same centrifuge 1ml of ammonium
oxalate solution is added mixed and allowed to stand for at least 30 minutes. The solution is
centrifuged for 10 minutes the supernatant is removed carefully without loss of the
precipitated. The precipitate is washed with 3ml of liquid ammonia for the centrifuged and
decanted and supernatant. It is washed with 3ml of distilled water and the supernatant is
removed as before after centrifuging. To the precipitate 2.0ml of 1N sulphuric acid is added
along the sides rotating the tubes so that the precipitate is washed down to the bottom the
tubes is placed is boiling hot water bath at 70-80°c. The calcium oxalate dissolved and the hot
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Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
solution is titrated against the standardized potassium permanganate till the end point is the
appearance of permanent pale pink colour a duplicated is conducted simultaneously.
A blank is conducted by taking 2.0ml of 1N sulphuric acid. Heated and against the
standardised potassium permanganate, until the end point is reached.
Determination of Moisture Content[10]
Moisture content was determined using standard method. Fresh seeds kept in a pre weighed
watch glass and dried at 150°C over night, in oven. The sample with watch glass cooled to
room temperature in a desiccator before weighing. The weight loss in sample, regarded as
moisture content of the sample.
Determination of Ash Content[11]
According to the method, 10g of each sample was weighed in a silica crucible. The crucible
was heated in a muffle furnace for about 3-5h at 600°C. It was cooled in a desiccator and
weighed to completion of ashing. To ensure completion of ashing it was heated again in the
furnace for half an hour more cooled and weighed. This was repeated consequently till the
weight become constant (ash become white or grayish white).
Determination of Vitamin B[12]
5gms of the individual sample was extracted with 100 ml of 50% ethanol solution and shaken
for 1 hr. This was filtered into a 100 ml flask; 10 ml of the extract was pipetted into 50 ml
volumetric flask. 10 ml of 5% potassium permanganate and 10 ml of 30% H2O2 were added
and allowed to stand over a hot water bath for about 30 min. 2 ml of 40% Sodium sulphate
was added. This was made up to 50 ml and the absorbance was measured at 510 nm in a
spectrophotometer.
Estimation of Vitamin-C[13]
Pipetted out 10-100 standard dehydro ascorbic solution into a series of tube. Similarly
pipetted out different aliquots (0.1-2ml) of brominated sample extract made upto the volume
in each tube to 3ml by added distilled water. Added 1ml of DNPH reagent followed by 1-2
drops of thiourea to each tube set a blank as above but with water in place of ascorbic acid
solution, mixed the contents of the tubes throughly and incubated at 37°C for 3 hours. After
incubation dissolved the orangered osazone crystal formed by added 7ml of 80% sulphuric
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Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
acid measured absorbance at 540nm plotted. A graph ascorbic acid concentration versus
absorbance and calculated the ascorbic content in the sample.
Estimation of Vitamin D[14]
To an accurately weighed amount of the sample (0.300mg), add 0.1 g of hydroquinone, 25 ml
of 0.5 N potassium hydroxide solution in ethanol and boil the mixture under reflux for 20
min. Cool, then add 50ml of water and extract with three 30-ml amounts of peroxide-free
ether. Wash the combined ethereal extracts with 20 ml of water, then with 20 ml of 0.5 N
aqueous potassium hydroxide and finally with successive volumes, each of 20 ml. Filter the
ethereal solution through a small plug of cotton-wool, wash the plug with two 10-ml volumes
of peroxide-free ether and evaporate the combined ethereal filtrate and washings to dryness
on a water-bath, at 50 ºC. Then dissolve the residue in 2 ml of methanol and 0.6 ml of
anisaldehyde reagent and 1.25 ml of analytical-reagent grade sulphuric acid, which is added
drop wise with constant agitation. Allow the mixture to stand for 1 min, then dilute to volume
with 10 per cent. V/V sulphuric acid in methanol and set it aside for 20 min. Measure the
absorbance of the greenish-blue colour at 685 nm against a blank solution similarly treated.
Estimation of Vitamin E[15]
To 500mg of sample, 1.5ml of ethanol was added, mixed for 15 minutes and centrifuged. The
supernatant was dried at 80oC, to this 0.2ml of 2,2'dipyridyl and 0.2ml of ferric chloride
solution were added, mixed well and 4.0ml of butanol was added. The developed red colour
was read at 520nm.
RESULTS AND DISCUSSION
Chickpea (Cicer arietinum L.) seed is an important and cheap source of legume protein
which can be used as a substitute for animal protein because their supply is limited and
expensive. The protein content of chickpea seed is highly variable and determined by both
genetic and environmental factors.[16]
Table1: Phytochemical Analysis of Raw, Soaking and Pressure-Cooked Seeds of Cicer
Arietinum (L.)
S.No Phytochemicals Raw seed Soaking seed Pressure-cooked seed
1. Alkaloids + + +
2. Carbohydrates + + +
3. Glycosides - - +
4. Protein + + +
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Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
5. Amino acid + + +
6. Phytosterols - - -
7. Tannins + + +
8. Phenols + + +
9. Flavanoids + + +
10. Coumarins + + +
11. Saponin + + +
12. Guinone - - -
13. Cardiac glycosides + + +
14. Terpenoid + + +
15. Phlobatannins - - +
16. Anthraquinones - - -
17. Steroids & Phytosteriods + + +
Table1 shows the presence of phytochemicals like alkaloids, carbohydrate, protein, tannins,
phenols, flavanoids, coumarins, saponins, cardiac glycosides, terpenoids, steroids and
phytosterols in raw, soaking and pressure-cooked seeds of Cicer arietinum (L.). At the same
time the pholabatannins was only found in the pressure-cooked seeds and not in the raw,
soaking seeds of water extract. Glycosides, phytosterols, Guinone, anthraquinones are not
found to be raw, soaking and pressure-cooked seeds of Cicer arietinum (L.).
Phytochemical analysis conducted on the seed extracts revealed the presence of constituents
while are known to exhibit medicinal as well as physiological activities.[5]
The phenolic
compounds are one of the largest and most ubiquitous groups of plant metabolites.[17]
They
possess biological properties such as antiapoptosis, antiaging, anticarcinogen,
antiinflammation, antiatherosclerosis, cardiovascular protection and improvement of
endothelial function, as well as inhibition of angiogenesis and cell proliferation activities.[18]
Several studies have described the antioxidant properties of medicinal seed which are rich in
phenolic compounds.[19]
Tannins bind to proline rich protein and interfere with protein synthesis. Flavanoids are
hydroxylated phenolic substances known to be synthesized by seed in response to microbial
infection and they have been found to be antimicrobial substances against wide array of
microorganism in vitro. Saponins have the property of precipitating and coagulating red
blood cells. Some of the characteristics of saponins include formation of foams in aqueous
solution, hemolytic activity, cholesterol binding properties and bitterness.[20]
Alkaloids have been associated with medicinal uses for centuries and one of their common
biological properties is their cytotoxicity. Steroids and triterpenoids show the analgesic
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Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
properties and responsiple for central nerrous system activity. Terpenes are very important
group of organic compounds that have been reported as potent drugs used in treatment of
wide range of ailments.[21]
Table2: Proximate Composition of Raw, Soaking and Pressure-Cooked Seeds of Cicer
Arietinum (L.)
S.NO Compounds Raw seeds Soaking seeds Pressure-cooked seeds
1. Carbohydrate 42.2±1.93 40±7.93 45.8±3.77
2. Protein 31.4±2.73 36.4±6.30 43.4±2.12
3. Iron 12.72±0.91 14.8±0.42 22±0.27
4. Calcium 29.3±0.35 30.2±0.02 32.0±0.22
5. Moisture 8.7±0.06 3.41±0.05 6.2±0.29
6. Ash 3.72±2.34 2.9±2.73 3.1±2.54
7. Vitamin C 0.69±0.01 0.83±0.05 0.91±0.04
8. Vitamin B 0.56±0.03 0.69±0.04 0.83±0.05
9. Vitamin D 0.09±0.06 0.14±0.01 0.16±0.01
10. Vitamin E 0.37±0.02 0.43±0.03 0.47±0.03
Figure1: Proximate Composition Of Raw, Soaking And Pressure-Cooked Seeds Of
Cicer Arietinum (L.)
The amount of carbohydrate were found to be higher in pressure-cooked seeds (45.8±3.77)
than soaking (40±7.93) and raw (42.2±1.93) seeds. Crude carbohydrates of pressure-cooked
seeds were greater than raw and roasted seeds. The energy value of pressure-cooked seeds
exceeded than raw and roasted seeds and was higher than commonly cultivated pulses.[22]
The protein content of pressure-cooked seeds (62.2±2.12) were higher than that of raw
(43.4±2.73) and soaking seeds (36.4±6.30). Microwave cooking and pressure cooking do not
affect the nutrient composition of legumes. Cooking treatments significantly decreased the
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Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
non-protein nitrogen, ash and fat contents. These decreases might be attributed to their
diffusion into cooking water.[23]
Crude fiber was significantly increased by cooking
treatments. This increase could have been due to protein-fiber complexes formed after
possible chemical modification induced by the soaking and cooking of dry seeds.[24]
Iron content of the pressure-cooked (22±0.27) seeds was comparatively higher than raw
(12.72±0.91) and soaking (14.8±0.42) seeds. Cooking had a significant effect on the mineral.
A significant improvement in availability of iron was also observed on cooking treatments
compared to the unprocessed treatment, which may be due to leaching out of antinutrients.[25]
Little variation in ash and moisture content of raw, soaking, and pressure-cooked seeds. Ash
content of pressure-cooked seeds was less than raw and soaking seeds.[26]
The decrease ash
content of pressure-cooked seeds was related to that fact that mineral or ash content of
vegetative tissues are positively related.[27]
The moisture content of the cooked pulses was
higher than the pressure-cooked pulses due to absorption of water during cooking which had
a dilution effect on all other nutrients.[28]
The amount of calcium content of Cicer arietinum (L.) Brown seeds averages pressure-
cooked (32.0±0.2) soaking (30.2±0.02) and raw (29.3±0.35) respectively. Pressure-cooking
drained most of the minerals from seeds except calcium, copper, zinc and manganese that
were similar in soaking and pressure-cooked seeds. The mineral content of raw and processed
seeds of the chickpea does not meet the recommended dietary allowance.[29]
Figure2: Estimation of Vitamin B, Vitamin C, Vitamin D and Vitamin E of Raw,
Soaking And Pressure-Cooked Seeds Of Cicer Arietinum (L.)
Vitamin content in raw, soaking and pressure-cooked seeds are presented in table 2 (figure 2).
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Sumathi et al. World Journal of Pharmacy and Pharmaceutical Sciences
Riboflavin, thiamin, niacin and pyridoxine in chickpea seeds were significantly reduced by
cooking treatments. These losses were probably due to a combination of leaching and
chemical destruction. The losses by microwave cooking were smaller than those obtained
with boiling and autoclaving.[30]
Cicer arietinum is good source of folic acid and coupled with more modest amounts of water
soluble vitamins like riboflavin, pantothenic acid and pyridoxine and these levels are similar
or higher than that observed in other pulses. Chickpeas contain several antioxidant vitamins
and properties that helps to clean the free radicals.[31]
The improvement in vitamin retention by microwave cooking may have been the result of
shorter cooking time compared to boiling and autoclaving. The sensitivity of vitamins to loss
from cooking was, in descending order: pyridoxine, riboflavin, thiamin and niacin.[30]
Similar
results were obtained for cooked kidney beans and carrot. Boiling resulted in a greater loss
for each vitamin compared to the other cooking treatments.[32]
CONCLUSION
Legumes are considered important and in expensive protein and dietary fibre sources in
human nutrition generally processing methods of chickpea is cooking at atmospheric
pressure, pressure cooking or microwaving. Two most commonly used cooking methods by
majority of Indian families namely, pressure cooking and boiling were employed.
Chickpea in future is potential protein source for humans and it is the first study on the
comparative nutritional quality of raw, soaking and pressure-cooked seed of chickpea. Some
important phytochemicals such as Alkaloids, carbohydrate, Protein, Amino acid, Tannins,
phenols, flavanoids, coumarins, saponin, cardiac glycosides, terpenoid, steroids and
phytosterols present in chickpea, plays crucial role of their beneficial effects. It also reduces
the risk of cardiac vascular diseases, glycemic index, blood pressure and prostate cancer. As
show in this study, raw, soaking and pressure-cooked method affect the nutritional quality of
chickpea. Pressure-cooked form of chickpea exceeds in carbohydrate, protein, iron, calcium
and vitamins than raw and soaked seeds. Pressure cooking method, when done under
controlled time and temperature usually improves the protein quality and digestibility and
inactivates protease and amylase inhibitors as well as lectins and increase the dietary fibre.
From the above study it is quality clear that cooking chickpea by pressure-cooked method not
only saves time but also retains the most nutritive value.
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