nutritional quality of raw and soaking compared to

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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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.


670


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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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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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


675


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


676


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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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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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


679


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).


680


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.


681


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