“STUDY OF NUTRIENT CONTENTS OF COMMON CHILIES IN

COMPARISON ON WITH HYBRID CHILI”

INTRODUCTION

Cayenne or Capsicum derives its name from the Greek, 'to bite'.

The dried fruit is a powerful local stimulant with no narcotic effect, it is

most useful in atony of the intestines and stomach.

Capsicum is the most pronounced, natural and ideal stimulant

known in the entire materia medica. It cannot be equaled by any known

agent when a powerful and prolonged stimulant is needed, as in

congestive chills, heart failure, and other conditions calling for quick

action. The entire circulation is affected by this agent and there is no

reaction. Capsicum increases the power of all other agents, helps the

digestion when taken with meals, and arouses all the secreting organs.

Whenever a stimulant is indicated, Capsicum may be given with the

utmost safety. It is the only natural stimulant worth considering for

diarrhea, dysentery with bloody mucus, stools, and offensive breath.

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The potent, hot fruit of cayenne has been used as medicine for

centuries. It was considered helpful by herbalists for various conditions

of the gastrointestinal tract, including stomach aches, cramping pains,

and gas. Cayenne was frequently used to treat diseases of the circulatory

system. It is still traditionally used in herbal medicine as a circulatory

tonic (a substance believed to improve circulation). Rubbed on the skin,

cayenne is a traditional, as well as modern, remedy for rheumatic pains

and arthritis due to what is termed a counterirritant effect. A

counterirritant is something that causes irritation to a tissue to which it is

applied, thus distracting from the original irritation (such as joint pain in

the case of arthritis).

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Capsaicin

The compound that is the active ingredient in chili peppers, which

allows them to be so valuable culinarily and as weapons, is called

capsaicin. Capsaicin (8-methy-N-vanillyl-noneamide) is the primarily

capsaicinoid. Capsaicin is what produces the "burning" sensation felt

when a chili pepper is eaten.. Capsaicin, a colorless crystalline substance,

was first synthesized in 1930. Capsaicin has the chemical formula

C18H27NO3.

Capsaicinoids are natural substances produced by chili peppers. Of

the capsaicinoid fraction, capsaicin (48.6%) is quantitatively followed by

6,7-dihydrocapsaicin (36%), nordihydrocapsaicin (7.4%),

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homodihydrocapsaicin (2%), and homocapsaicin (2%) . Capsaicinoids

and capsaicin are collectively found in amounts of 0.1% to 1%, with

quantities varying according to soil and climate. The most important

constituents of Capsicum are pungent phenol compounds (0.05 - 1.5 %)

Some other constituents include carotenoids (capsanthin, capsorubin,

carotene, lutein etc.). There is also a minute quantity of a liquid alkaloid,

a saponin capsicidin and a fixed oil. Capsicum also contains up to 0.2%

ascorbic acid. The vitamin and mineral content (per 100 gm) is calcium

(29 mg), phosphorus (78 mg), iron (1.2 mg), potassium (374 mg),

thiamine (0.22 mg), riboflavin (0.36 mg), and niacin (4.4 mg). Capsicum

is also rich in fats (9-17%) and protein (12-15%).

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It is has high solubility in alcohol, but very low solubility in water.

It should be noted that since capsaicin is an oil, its lipophilic property

allows it to be solvable in fat. Capsaicin melts at 65 degrees Celsius, and

boils at 210-220 degrees Celsius (0.01 torr). This molecule is not

primarily found in the seed as mistakenly believed by many, but in the

white "ribs" of chili peppers, its placenta, that runs down the sides and

the middle of the chili peppers. Capsaicin is also unevenly distributed

throughout the flesh of chili peppers, so certain areas of a chili pepper

can be spicier than others. This compound is believed to be a part of the

defense mechanism of Capsicum plants.

There is confusion regarding the classification of Capsicum

species. Currently, all varieties of mild and hot peppers (not to be

confused with black and white pepper derived from Piper nigrum and

related species) are considered as the fruits of a single species, C.

annuum and its many varieties, or of 2 species, C. annuum and C.

frutescens. Current practice is to classify the pungent varieties of pepper

(chile peppers or cayenne peppers) as C. frutescens, and the milder-

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flavored sweet peppers (bell peppers, sweet peppers, green peppers) as

varieties of C. annuum, however, most botanists agree that they should

properly be regarded as varieties of a single species.

Classification

Scientific name : Capsicum annuum L.

Family : Solanaceae

Common Names : Capsicum, chili pepper, hot pepper, cayenne, red

pepper, tabasco paprika pepper, sweet pepper, bell

pepper, green pepper.

Scientific name : Capsicum frutescens L.

Family : Solanaceae

Common name : Capsicum(Cayenne,Red Pepper)

Synonyms : Red Pepper, Bird Pepper, African Bird

Pepper ,African

    Pepper,Chillies ,African Chilies,

Part Used : Fruit, ripe and dried.

Habitat : Zanzibar,but now grown in most tropical and sub-

tropical countries.

Therapeutic Applications of Capsicum:

6

Despite the widespread use of chili peppers in the diet, little is

known about the pharmacological activities of capsaicin in humans.

Cayenne was frequently used to treat diseases of the circulatory system.

It is still traditionally used in herbal medicine as a circulatory tonic.

a)Primary Uses: Arthritis, Bleeding, Blood Pressure (high/low),

Bronchitis, Poor circulation, Colds, Congestion, Diabetes, Fatigue,

Gastric Disorders, Heart Problems, strokes, Kidney Problems, Lung

Disorders, Phlebitis, Rheumatism, Shock, Tumors, Sore throat, Varicose

veins, Ulcers, anti-inflammation.

b)Secondary Uses: Arteriosclerosis, Asthma, Blood Impurities,

Bruises, Burns, Fevers, Gas, Infections, Jaundice, Malaria,

Mucus/excessive, Pain, Pancreatic Problems, Pus Discharge, Sinus

Problems, Skin Disorders, Spasms, Sunburns, Wounds.

   The Healing Power of Capsicum describes remedies using

capsicum alone or in mixtures with ordinary items like lemon, vinegar,

olive oil, honey, garlic , aspirin and more for common health problems

like Angina, Clogged Arteries, Bruises and Sprains, Colds and Flu,

Cough, Diabetes, High Cholesterol, Headache, Neuralgia and

Obesity. 

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Hence chilies were highly nutritive and widespread in use ,

we are aimed to determine the nutritive content of chili in comparison

with hybrid chili.

OBJECTIVE

To analyze the Nutrient contents in common chilies and hybrid chili.

Quantitative analysis of Biomolecules

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Total carbohydrate Reducing sugar Starch Protein

Estimation of Vitamin- C

Ascorbic acid

Bioactive profile of pigments

Chlorophyll a Chlorophyll b

Determination of secondary product

Phenol

REVIEW OF LITERATURE

Archeologists estimate that in Mexico, Capsicum was used as a

food as long as 9,000 years ago (Rumsfield and West, 1991). The

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medicinal use of a number of Capsicum species, including C. annuum by

the Mayans, is described in Chichewicz and Thorpe (1996). They include

the use of roots, leaves, as well as the fruits in applications for infections,

fresh burns, respiratory complaints, earaches, and sores.

Regular ingestion of hot red pepper is recommended by some

authors for anorexia, hemorrhoids, liver congestion, varicose veins, and

vascular conditions (Duke, 1985). Pedersen (1994) states that "the most

striking use of Capsicum is as a catalyst herb in nearly every herbal

combination conceivable."

Capsicum was introduced into Britain from India in 1548, and

Gerard mentioned it as being cultivated in his time. The plant was

described by Linnaeus under the name of C. frutescens proper. This

species appeared in Miller's Garden Dictionary in 1771.

Chemical constituents

a) Nitrogenous compounds:

  The most potent and predominant chemical entity in Capsicum is

capsaicin (0.14%) (Cordell and Araujo, 1993). A series of homologous

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branched- and straight-chain alkyl vanillylamides, collectively known as

capsaicinoids. Of the capsaicinoid fraction, capsaicin (48.6%) is

quantitatively followed by 6,7-dihydrocapsaicin (36%),

nordihydrocapsaicin (7.4%), homodihydrocapsaicin (2%), and

homocapsaicin (2%) (Duke, 1985). Capsaicinoids and capsaicin are

collectively found in amounts of 0.1% to 1%, with quantities varying

according to soil and climate (Rumsfield and West, 1991).

    Capsaicin, a colorless crystalline substance, was first synthesized

in 1930. Capsaicin has been studied since the mid-19th century and its

structure is elucidated as 8-methyl-6-nonenoyl vanillylamide (Cordell

and Araujo,1993).

The crude extract of Capsicum fruits, known as Capsicum

oleoresin, contains at least 100 different volatile chemical constituents,

and therefore may function in differing ways from pure capsaicin. Thus,

it is important to distinguish between studies using capsaicin and those

employing Capsicum oleoresin` (Cordell and Araujo, 1993).

b) Steroids

Other parts of the plant contain steroidal alkaloid glycosides

(solanine, solanidine, solasodine) (Newall et al., 1996). The seeds contain

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the steroidal glycosides capsicoside A through D, all furostanol

glycosides (Yahara et al., 1994)

c) Other Constituents:

C. annuum is rich in carotenoid pigments, including capsanthin,

capsorubrin, carotene,luteine, zeaxanthin, and cucurbitaxanthin A)

(Leung and Foster, 1996; Hornero-M¨¦ndez and Manguez-Mosquera,

1998). Capsicum is also rich in fats (9-17%) and protein (12-15%)

(Leung and Foster, 1996) and is an excellent source of vitamin C (~370

mg/100 g) and vitamin A (77,000 IU/100 g, equivalent to 7,700 RE/100

g) (Ensminger et al., 1993). Volatile oils are present as a trace

component, including over 125 individual constituents, 24 of which have

been identified (Marsh, 1977).

History of Capsaicin:

P.A. Bucholtz in 1816 first discovered that the pungent

principle of peppers could be extracted from the macerated pods with

organic solvents. In 1846, L. T. Thresh reported in Pharmacy Journal that

the pungent principle could be extracted in a crystalline state. It was

Thresh who named the substance Capsaicin. In 1878, the Hungarian

medical scientist Endre Hogyes extracted Capsaicin, which he called

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capsicol, and discovered that it stimulated the mucous membranes of the

mouth and stomach and increased the secretion of gastric juices.

Capsaicin was first synthesized in 1930 by E. Spath and F.S. Darling. It

has virtually no odor or flavor, but it is one of the most pungent

compounds known, detectable to the palate in dilutions of one to

seventeen million. It is slightly soluble in water, but very soluble in

alcohol, fats, and oils.

The word capsaicin actually describes a complex of related

components named capsaicinoids by Japanese chemists S. Kosuge and Y.

Inagaki in 1964. Capsaicinoids are the chemical compounds that give

chili peppers their bite.

Mode of Action

  The Dispensatory of the United States of America, 23 ed.,

1943, states that "Capsicum is a powerful local stimulant, producing,

when swallowed, a sense of heat in the stomach, and a general glow over

the body without narcotic effect." Capsicum sp. are known to be very

strong local stimulants in the circulatory system. In studies performed on

female rabbits, capsaicin, the major constituent of cayenne, has been

13

shown to significantly lower both plasma cholesterol and triglycerides,

but even more important, lower the LDL-HDL ratio.

Besides its action as a local stimulant, Capsicum acts as a

powerful stimulant to the digestive tract when taken internally. Capsicum

is a gastric,stimulant,stomachic, carminative and an internal

tonic.Cayenne is of particular value for atonic gastric dyspepsia for atony

of the stomach and intestines and has been widely used to treat

flatulence. Capsicum is used externally as local counterirritant for

rheumatism, neuralgia, arthritis, chilblains and lumbago.Cayenne has

been used in North America, Europe, China and in India. It was listed in

Gerard (1597), Lewis (1769) and Comfort (1853). It can also be found in

Aztec herbals (1552).(Crellin, J.K. and Philpott, J., Herbal Medicine:

Past and Present (Vol. II), Duke University Press, London, 1990, p. 142.)

Functions

a) Cardiovascular and Circulatory Functions:

Yamato et al. (1996) showed that capsaicin produced a marked

concentration-dependent decrease in the amplitude, the rate of rise, and

the rate of relaxation of the contractile tension of rat ventricular papillary

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muscles; however, the half-life of the relaxation and the time to peak

tension were only slightly effected.

b) Digestive and Gastrointestinal Fuctions:

The authors note, however, that while chili pepper, cayenne

pepper and paprika may loosen cell contacts to increase permeability of

the intestinal epithelium, other spices (bay leaf, black pepper, and

nutmeg) were found to decrease permeability (Jensen-Jarolim et al.,

1998).

The stimulatory effect of orally administered capsaicin on

gastric acid secretion and mucosal blood flow was studied in rats using

amounts roughly equivalent to a normal Thai diet. Capsaicin was noted to

have a protective effect on gastric mucosa of ethanol-induced gastric

lesions in rats (Uchida et al., 1991).

c) Immune Functions:

(i) Carcinogenicity/ Mutagenicity

   A full review of capsaicin's carcinogenic and anticarcinogenic

potential (Suhr and Lee, 1996) provided theoretical evidence for both

effects from capsaicin. Ernst and Barnes (1998) refer to this study with

15

the comment that "Taken orally orally in regular high doses it may act as

a carcinogen and could promote gastric cancer, but in low doses it seems

to have anticarcinogenic activity." Duke (1985) points out that the low

incidence of gastric cancers in Latin America suggests that hot pepper,

with its many constituents, may be anticarcinogenic.

A 10% Capsicum, protein-deficient diet fed to rats led to a

54% increase in the incidence in hepatomas, suggesting that capsaicin

may contribute to the development of liver cancer. Nalini et al. (1998)

report that rats fed a diet containing red chili (8 mg/day/100 g body

weight) alone or with a carcinogenic substance (DMH, 1,2-dimethyl

hydrazine, 20 mg/kg, s.c.) in addition to the red chili showed a tumor

incidence of 83.3% and 93.3%, respectively.

Chili prepared by sun-drying, salting and deep-frying in

groundnut oil contains a high amount of carcinogenic 3,4-

benzo(a)pyrene. Long-term feeding studies in male mice with chili so-

prepared (100 mg/day) added to a laboratory rodent diet found that

whereas none of the controls developed tumors, the chili group showed a

35% incidence of adeno carcinoma in the abdomen over 2 yrs.

The authors comment that their results suggest the high

incidence of gastric cancer among the male population of Madras may

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owe to the contributing factor of salted, sundried and oil-fried red chili

(Balachandran and Sivaramkrishnan, 1995).

    Park et al. (1998) found tumor-promoting activity from

capsaicin lacking in a 2-stage skin carcinogenesis model in mice. Instead,

capsaicin, when administered at the same time as a tumor promoter (12-

O-tetradecanoylphorbol-13-acetate), was found to inhibit mouse skin

carcinogenesis.

(ii) Immune modulation:

The immunomodulatory effects of capsaicin are

varied and may be related to interactions with the neuropeptides

somatostatin and SP, a peptide made up of 11 amino acids and found

throughout the body in nerve cells and certain endocrine cells in the gut

In addition, increased vascular permeability promotes the

local delivery of both the protein and cellular components of adaptive

immunity, so that SP could augment the activity of T lymphocytes that

accumulate at the site of reaction (Payan et al., 1984).

(iii) Pharmacokinectics:

 With respect to topical applications of capsaicin, it

has been estimated that assuming 100% of a topically-applied dose is

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absorbed into the body, an application of 90 g capsaicin (2 tubes of

cream, 0.025% capsaicin) per week would result in a daily exposure of

0.064 mg/kg capsaicin for a 50 kg person. This represents less than 10%

of the dietary intake of a typical Indian or Thai diet (Rumsfield and West,

1991).

Safety Profile

a) Contraindications

Capsicum has been cited as contraindicated in topical

applications on damaged skin and near the eyes, and for internal use by

individuals who are sensitive to the herb and may in some cases develop

gastrointestinal irritations. Further contraindications in the German

Commission E are found in topical applications by individuals who are

allergic (sensitive) to Capsicum (Blumenthal et al., 1998).

  Lynn (1990) describes that relatively mild capsaicin topical

treatment has been reported to worsen contact dermatitis and allergic

contact dermatitis in some individuals. Willard (1991) considers use of

Capsicum contraindicated in ulcers and chronic bowel irritation states.

  Excessive doses have been said to cause severe irritation of

mucous membranes, nausea, vomiting and diarrhea. The active, pungent

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principle of Capsicum can be an irritant to the eyes and to tender skin,

producing a strong burning sensation.

 b) Pregnancy

No documented cases of adverse effects from use of capsaicin

during pregnancy have been found. Brinker (1983) states that Capsicum

oleoresin was found to be a uterine stimulant in animals.

 c) Side Effects

Initial topical application of capsaicin creams results in burning

sensations for most but not all people, which lessens or disappears with

repeated applications. Erythema often accompanies the burning,

sometimes with rash. Coughing and sneezing from aerosolized particles

from dried cream residues has also been noted in some studies.

Accidental contamination of other body parts, particularly the eyes,

mouth or perineal regions, can occur without careful hand-washing or the

use of rubber gloves for cream application (Mitchell and Rook, 1979). In

a controlled study, Jones et al. (1997) found that cool tap water was more

effective at providing immediate relief from the pain of chili burns of the

hands than room temperature vegetable oil. Immersing the hands in the

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vegetable oil provided significantly better long-term pain relief, provided

the hands were immersed in the oil for at least 1 hour.

Capsaicin and capsaicinoids are strongly irritating to mucous

membranes and can produce dermatitis. Inhalation can produce allergic

alveolitis (Mitchell and Rook, 1979). Oral use of Capsicum and its

extractives may cause gastrointestinal irritation, though it does not inhibit

the healing of duodenal ulcers and does not need to be avoided by

persons with such a condition (Leacock, 1985).

Source:

The samples (common chilies and the hybrid chili) were collected

and weighed in required amount then it was smashed well using motor

and pestle then it was homogenized using the homogenizer and dissolve

in respective solvents. Then the mixture was centrifuged the supernatant

obtained was taken and the quantitative estimation was proceeded and

their values were read at respective optical densities.

MATERIALS AND METHODS

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ESTIMATION OF TOTAL CARBOHYRATE

(Anthrone method )

Principle:

Carbohydrate was first hydrolysed into simpler sugar using dilute

hydrochloric acid. In hot acidic medium glucose in dehydrated to

hydroxyl methyl funeral. This forms a green colored products with

phenol and at colorimetrically 490nm.

Reagents:

1. 2.5 hydrochloric acid

2. anthrone 2%

3. sulphuric acid965%

4. standard glucose solution (Stock standard)

5. Working standard

6. Source

100mg of the sample was weighed and it was hydrolysed by

keeping in a boiling water bath for 3 hours with 5ml of 2.5 N

Hydrochloric acid and cooled at room temperature. They were

neutralized with solid sodium carbonate until the effervescence

ceases.

Procedure:

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The volume was made up to 0.2, 0.4, 0.6, 0.8 and 1ml of the

working standard were piptted out in a series of test tubes. 0.2 ml of the

sample solution was pipetted out in another test tube. The volume was

made up to 1ml with distilled water in all the tubes. 5ml of 96% sulphuric

acid was added and mixed well. After 10 min the contents in the tubes

were shaken and placed in a water bath at 25-30 0C for 20 min. 4 ml of

anthrone reagent was added and heated for 8 minutes in boiling water

bath cooled rapidly and read at 680nm. The total carbohydrate present in

the sample was calculated using the standard graph.

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ESTIMATION OF REDUCING SUGAR

(Somogyi, 1952)

Principle:

The reducing sugar when heated with alkaline copper tartarate reduce

the copper from the cuprous state and thus cuprous oxide was formed. When

cuprous oxide was treated with arsenomolybdic acid, the reduction of

molybdic acid to molybdenum blue takes place. The blue color developed

was compared with a set of standard in a colorimeter at 620nm.

Reagents:

1. Alkaline copper tartarate:

A. Dissolved 2.5 gm of anhydrous sodium carbonate, 2 gm of sodium

bicarbonate, 2.5 gm of potassium sodium tartarate and 20 gm of

anhydrous sodium sulphate in 80 ml of water and made up to 100

ml

B. Dissolve 15gm of copper sulphate in a small volume of distilled

water. Added one drop of sulphuric acid and made up to 100ml.

Before use , 4ml of B and 96ml of solution A were mixed.

2. Arsenomolybdate reagents:

Dissolve 2.5gm of ammonium molybdate in 45ml of water.

Added 2.5ml of sulphuric acid and mixed well. The 0.3gm of disodium

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hydrogen arewsnate was dissolved in 25ml of water mixed well and

incubate at 370C for 24 to 48 hours.

3. stock standard glucose solution

4. working standard

Procedure:

Weighed 100mg of the sample and the sugar were extracted with hot

80% ethanol twice . the supernatant was collected and evaporated by

keeping it on a water bath at 800 C. 10ml of water was added and the sugar

were dissolved 0.2, 0.4, 0.6, 0.8 and 1 ml of working standard solution and

0.2mml of the extract solution was pipetted out into a series of test tubes.

The volume in all tubes were made up to 2ml with distilled water was kept

as blank. 1ml of alkaline copper tatarate reagent was added to all tubes. The

tubes were kept in boiling water bath for 10 minutes. The tubes were cooled

and 1 ml of arsenomolybdic acid reagent was added to all the absorbance of

blue color was read at 620nm after 10minutes. From the graph the amount of

reducing sugar present in the sample was calculated.

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ESTIMATION OF STARCH

( Mc Cready et al., 1950

Principle :

Perchloric acid was found to be a very efficient solvent for starch.

The residue rich in starch was solubilised with perchloric acid and were

determined colorimetrically with anthrone reagent.

Reagents:

1. 0.2% Anthrone reagent: 0.2 gm of anottrone dissolved in 100ml of

concentrated sulphuric acid.

2. 80% ethanol

3. 52% perchloric acid.

Extraction:

To 100 mg of powered few drops of 80% ethanol and 5ml of water

were added and stirred well. The soluble sugars were eliminated with

80% hot ethanol by repeated extraction. To the sugar free residue added

3ml of water and 6.5ml of 25% perchloric acid was added with constant

stirring. The tube was kept cool by immersing in an ice for 15 minutes.

Then 5ml of distilled water was added, mixed well and centrifuged. This

residue was then extracted with perchloric acid to ensure complete

25

extraction of starch. The combination extract was diluted to 100 ml using

distilled water.

Procedure :

0.5ml of the above solution was made up to 1ml with distilled

water to which 5ml of freshly prepared anthrone reagent was added and

cooled in an ice bath for 10 minutes. Blank and standard were treated in

the same way. The color intensity was measured in the wavelength of

620m against blank. Starch content was expressed as mg of starch per gm

dry weight.

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ESTIMATION OF PROTEIN

( Lowry et al.,1951)

Principle:

The blue colour was developed by the reduction of the

phosphomolybdie phosphotungstic components. The Folin-ciocalteau

reagent by the amino acids tyrosine and tryptophan present in the protein

plus the colour developed by the biuret reaction of the protein with the

alkaline cupric tartarate was measured in the Lowry’s method.

Reagents:

1. Reagent A : 2% sodium carbonate in 0.1N sodium hydroxide.

2. Reagent B : 0.5% copper sulphate (CuSO4: 5H20) in 1%

potassium sodium tartarate

3. Reagent C : Alkaline copper sulphate solution : Mixed 50ml of

A and I ml of B prior to use.

4. Reagent D : Folin- Ciocalteau Reagent :

Refluxed gently for 10 hours a mixture consisting of

100g sodium tungstate, 25gm sodium molybdate, 700 ml water,

50ml water and a few drops of bromine water. Boiled the mixture

for 15 min without condenser to remove excess bromine. Cooled,

diluted to 1 liter and filtered.

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5. Protien solution : weighed accurately 50 gm of Bovine Serum

Albumin and dissolve in distilled water and made up to 50 ml in

standard flask.

6. Working standard : diluted 10ml of the stock solution to 50ml with

distilled water in a standard flask.

Procedure:

Extraction of protein from sample:

Weighed 500mg of the sample and grinded well a pestle and

mortar in 10ml of the phosphate buffer. Centrifuged and used the

supernatant for estimation.

Estimation of protein :

Pipetted out 0.2, 0.4, 0.6, 0.8 and 1 ml of the working standard into

a series of test tubes. Pipetted out 0.1 of the sample extract in another

test tube. The volume was made up to 1 ml in all test tubes. A tube with 1

ml of water served as a blank. Added 5 ml of reagent C to all the tubes

including the blank. Mixed well and allowed to stand for 10 min then

0.5ml of reagent D was added , mixed well incubated at room

temperature in the dark for 30 min . Blue color was developed. Reading

were taken in spectrophotometer at 660 nm. Standard graph was drawn

and the amount of protein in the sample was calculated.

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ESTIMATION OF ASCORBIC ACID

Di-chloro- Indo-phenol Method

Principle

The dye which is blue in alkaline solution and red in acid solution

is reduced by ascorbic acid to a colorless compound as the same time

ascorbic acid is oxidized to dichloroascorbic acid. The reaction is

quantitative and specific for ascorbic acid in the pH range of 1-3.5.

Reagents:

1. 4% oxalic acid solution

2. Dye solution

3. Stock standard ascorbic acid

4. Working standard

5. Source

100 mg of sample was weighed and the juice was extracted

with 4% oxalic aid and make up to 1000ml with 4% oxalic acid.

Procedure:

Titration – I

Std Ascorbic acid Vs dye

5ml of std Ascorbic acid solution was pipetted out into a clean

conical flask and added 5ml of 4% oxalic acid. This was titrated against

29

dye solution taken in the burette. The end point was appearance of

permanent pale pink colour. The titration was repeated for concordant

value.

Dye factor = 0.5

Titer value

Titration – II

Test solution Vs dye

5ml of sample was pipetted out into a clean conical flask and 5ml

of 4% oxalic acid. It was then titrate against the dye solution taken in the

bottle. The end point was the appearance of permanent pale pink color.

30

ESTIMATION OF CALCIUM

(Clark and Collip)

Principle:

Both ionic and protein bound calcium is precipitated as calcium

oxalate by adding ammonium oxalate solution. The calcium oxalate

precipitated is dissolved in acid and titrated against standard potassium

permanganate at about 800C. Calcium is calculated from the titer value.

Procedure:

Pipette out 1 ml of the sample in a centrifuge tube. Add 3ml of

distilled water and 1ml of 4% ammonium oxalate. Mix the contents and

allow to stand for 30 minutes. For the calcium to precipitate after 30

minutes centrifuge for about 15 minutes at 300rpm discard the

supernatant add 3ml of 2% ammonia to the precipitate mixed well and

centrifuge. Repeat the process of centrifuging and discarding for 2-3

times.

To the washed precipitate add 2ml of 1N H2SO4 keep the tubes in

the beaker containing water and heat to about 80oc for contents in a hot

condition against 0.01N KMNO4. The end point is the appearance of faint

pink color which persists for a minute.

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ESTIMATION OF TOTAL PHENOL

(Bray and Thorp,1954)

Principle:

Estimation of phenol with folin-phnol reagents was based on the

reaction between phenol and an oxidizing agent phosphomolybdate

which results in the formation of a blue color. The intensity of color

developed was read colorimetrically at 650nm.

Reagents:

1. Folin phenol reagent

2. Sodium carbonate 20%

3. Catechol stock standard

4. Working standard was prepared by diluting 10ml of stock to

100ml.

Procedure:

1ml of the extract was pipetted out into a test tube and 1 ml of

folin-phenol following by 2ml of sodium carbonate were added. The

tubes was shaken and heated in a boiling water bath for exactly 1 min

then it was cooled under the tap water. The blue colour developed was

diluted to 25ml with distilled water and its absorbance was measured

calorimetrically at 650nm. The unknowns were read from a standard

32

curve made from different concentration of catechol. A blank containing

all the reagents minus plants extract was used to adjust the absorbance to

zero.

33

ESTIMATION OF CHLOROPHYLL

Principle:

Chlorophyll was extracted in 80% acetone and the absorption at

663 and 645nm were read in a spectrophotomer. Using the absorption

coefficient the amount of chlorophyll was calculated.

Reagents:

80% Acetone

Procedure:

1 gm of fresh capsicum were cut into small pieces and

homogenized in a motor using pestle using 80% acetone. Decanted and

filtered the supernatant through a funnel using Whatman No. 42 filter

paper. Added sufficient quantity of 80% acetone and repeated the

extraction. Transferred the contents from the motor to a funnel and

washed the brei with acetone until it become colourless. Pooled the

filtrates and made up the volume to 100ml in a volumetric flask.

Transferred,5ml of the extract into a 50ml volumetric flask and diluted by

making up the volume with 80% acetone. Measured the absorbance at

645 and 663nm for the determination of chlorophyll-a and chlorophyll-b

and total chlorophyll.

34

The chlorophyll content was calculated on the fresh weight basis

using the formula:

mg chlorophyll a/g tissue = 12.7 (A663) -2069(A645) x V

1000 x W

mg chlorophyll b/g tissue = 22.9 (A645)- 4.68(A663) x V

1000 x W

mg total chlorophyll b/g tissue = 20.2(A645) +8.02 (A663) x V

1000 x W Where,

A - Absorbance at specific wavelengths,

V - Final volume of chlorophyll extract in 80% acetone

W – Fresh weight of tissue extract

35

ESTIMATION OF CAROTENOIDS

Carotenoid extraction:

Total carotenoid was extracted from the muscles, exoskeleton

abdomen, hepato pancreas and gills. For extraction of carotenoids

solvent such as acetone lacetonitrile in the ratio of 95:5 was used. The

sample were homogenized well until the pigment was completely

extracted. The sample were centrifuged at 500rpm for 10 minutes and the

supernatant was allowed to condensed to a minimum volume in a water

bath at 40oc. the sample were stored in deep freezers from this sample

both quantitative and qualitative analysis were carried out.

Estimation of Total carotenoids

The extracted carotenoids from each sample was diluted to

appropriate volume and the optical density was measured. The same time

solvent used for extraction of carotenoids was used for dilution. After

dilution optical density was measured at 440nm.

Total carotenoids in the sample was estimated using the formula

C- Carotenoid content V- Volume of sample

D- Optical density F- Dilution factor

36

C = DVF

2500

RESULTS AND DISCUSSION

As we are using a large number of common chilies in our day to

day life, our “PROJECT STUDY OF NUTRIENT CONTENTS OF

COMMON CHILIES IN COMPARISON WITH HYBRID CHILI”

has its own use.

Of the common chilies pepper is found to be more nutritious in all

respect to the all the ingredients present in it, so when it is compared with

Hybrid chili the result obtained is as follows.

The fig: 1 shows that Total carbohydrate content is 98mg/dl in

pepper and while in hybrid chili it is only 3 mg/dl.

The fig: 2 shows that Total reducing sugar content in the common

chili is 150% in pepper where as the hybrid chili contain only 32%.

Fig: 3 shows that Starch content is 91 mg/dl in pepper while in

hybrid chili it is 3.5 mg/dl.

The fig: 4 shows that the Protein content is 160 mg/dl in pepper

where as the hybrid chili contain 80mg/dl.

The fig:5 shows that the Ascorbic acid content is 15mg/dl in

pepper while the hybrid contains 9.2mg/dl.

The fig: 6 shows that the Calcium content in the pepper is

28/mg/dl and 13mg/dl in the hybrid chili.

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The fig:7 shows that the Phenol content is 5 mg/dl in pepper and in

hybrid it is 3.5 mg/dl.

The fig:8(a) and fig: 8(b) shows that the chlorophyll a and

chlorophyll b content is more over equal in all the chilies.

Fig: 9 shows that the carotenoid content is found to be higher in

Dry chili, 0.002 mg/dl and least in Hybrid chili, 0.00032 mg/dl.

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Comparison of Nutrient contents in Chilies

Total Carbohydrate

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Total Carbohydrate (Anthrone method) 98 mg/dl 3 mg/dl 60 mg/dl 76 mg/dl 90 mg/dl

39

Total Reducing Sugar

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Total Reducing Sugar 150 % 32 % 40 % 30 % 35 %

40

Starch

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Starch 91 mg/dl 3.5 mg/dl 1.5 mg/dl 4.5 mg/dl 4 mg/dl

41

Protein

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Protein ( Lowry’s Method) 160 mg/dl 80 mg/dl 96 mg/dl 100 mg/dl 70 mg/dl

42

Ascorbic acid

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Ascorbic acid ( Di-chloro-indo-phenol method)

15 mg/dl 9.2 mg/dl 96 mg/dl 100 mg/dl 70 mg/dl

43

Calcium

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Calcium 28 mg/dl 13 mg/dl 14 mg/dl 12 mg/dl 8 mg/dl

44

Phenol

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Phenol 5 mg/dl 3.5 mg/dl 1.5 mg/dl 4.5 mg/dl 4 mg/dl

45

Clorophyll a

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Chlorophyll a 0.032 mg 0.033 mg 0.30 mg 0.009 mg 0.015 mg

46

Chlorophyll b

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Chlorophyll b 0.056 mg 0.056 mg 0.044 mg 0.020 mg 0.027 mg

47

Carotenoids

Nutritive contentsPepper (Black

pepper)

Capsicum (Bell

pepper)

Green Chili (Cayenne)

Dry Chili (Cayenne)

Pandi Chili (Pimento)

Carotenoids 0.001 mg 0.00032 mg 0.0004 mg 0.002 mg 0.0008 mg

48

CONCLUSION

The project “Study of Nutrient Content of Common Chilies in

Comparison with Hybrid Chili.” We came to the conclusion that common

chilies are more nutritious than the hybrid one and of them Pepper is

highly nutritious because it contains higher concentration of

Biomolecules such as Total protein, Reducing sugar, Starch, Protein etc.

mineral content is also higher in pepper. While analyzing the Vitamin

content that also showed positive result for pepper only. So from all these

we can say that pepper is highly nutritious.

While analyzing the pigment chlorophyll it is often more or less

equal in all chilies. But the Carotinoid content is some what higher in red

chili.

The secondary metabolite such as phenol was also found in more

amounts in the Pepper When compared with others and this is a notable

point because even though the chilies have good qualities like the

capability of wound healing, it also helps in some metabolic activities.

Due to the presence of higher concentration of Phenol content, the over

49

consumption of chilies like Pepper may lead to carcinogenic diseases and

other toxic effects.

Project work done by

Aneesh.D

Sreejith.S.Nair

Jones Kensy

Starlet Priya

Priya.L

Submitted to

Department of Biochemistry

Malankara Catholic College, Mariagiri

Kaliakkavila, Kanyakumari,

Tamil Nadu,

India-629 153

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