“STUDY OF NUTRIENT CONTENTS OF COMMON CHILIES IN COMPARISON ON WITH HYBRID CHILI”
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.
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).
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%),
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%).
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 milderflavored 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 Family : Capsicum annuum L. : Solanaceae pepper, tabasco paprika pepper, sweet pepper, bell pepper, green pepper. Scientific name Family Common name Synonyms ,African Pepper,Chillies ,African Chilies, Part Used Habitat : Fruit, ripe and dried. : Zanzibar,but now grown in most tropical and subtropical countries.
Common Names : Capsicum, chili pepper, hot pepper, cayenne, red
: Capsicum frutescens L. : Solanaceae : Capsicum(Cayenne,Red Pepper) : Red Pepper, Bird Pepper, African Bird Pepper
Therapeutic Applications of Capsicum: 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 Obesity.
Cholesterol, Headache, Neuralgia
Hence chilies were highly nutritive and widespread in use , we are aimed to determine the nutritive content of chili in comparison with hybrid chili.
To analyze the Nutrient contents in common chilies and hybrid chili. Quantitative analysis of Biomolecules 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 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.
a) Nitrogenous compounds: The most potent and predominant chemical entity in Capsicum is capsaicin (0.14%) (Cordell and Araujo, 1993). A series of homologous branched- and straight-chain alkyl vanillylamides, collectively known as capsaicinoids. Of the capsaicinoid fraction, capsaicin (48.6%) is quantitatively followed (7.4%), by 6,7-dihydrocapsaicin (2%), (36%), 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 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 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 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.)
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
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
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 soprepared (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
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 (12O-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
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
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
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).
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
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:
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.
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
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.
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
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.
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.
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.
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
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.
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.
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
curve made from different concentration of catechol. A blank containing all the reagents minus plants extract was used to adjust the absorbance to zero.
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.
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 V 1000 x W
mg chlorophyll b/g tissue = 22.9 (A645)- 4.68(A663) x
mg total chlorophyll b/g tissue = 20.2(A645) +8.02 (A663) x Where, A - Absorbance at specific wavelengths, V - Final volume of chlorophyll extract in 80% acetone W – Fresh weight of tissue extract
V 1000 x W
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=
C- Carotenoid content D- Optical density
V- Volume of sample F- Dilution factor
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 28/mg/dl and 13mg/dl in the hybrid chili.
in the pepper is
• 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.
Comparison of Nutrient contents in Chilies Total Carbohydrate
100 90 80 70 60 50 40 30 20 10 0
98 90 76 60
Pepper Capsicum Green chilli Dry chilli Pandichili
Nutritive contents Total Carbohydrate (Anthrone method)
Pepper (Black pepper) 98 mg/dl
Capsicum (Bell pepper) 3 mg/dl
Green Chili (Cayenne) 60 mg/dl
Dry Chili (Cayenne) 76 mg/dl
Pandi Chili (Pimento) 90 mg/dl
Total Reducing Sugar
160 140 120 100 80 60 40 20 0
Pepper Capsicum Green chilli Dry chilli Pandichili 35
Nutritive contents Total Reducing Sugar
Pepper (Black pepper) 150 %
Capsicum (Bell pepper) 32 %
Green Chili (Cayenne) 40 %
Dry Chili (Cayenne) 30 %
Pandi Chili (Pimento) 35 %
100 90 80 70 60 50 40 30 20 10 0 3.5 1.5 4.5 4 Pepper Capsicum Green chilli Dry chilli Pandichili
Pepper (Black pepper) 91 mg/dl
Capsicum (Bell pepper) 3.5 mg/dl
Green Chili (Cayenne) 1.5 mg/dl
Dry Chili (Cayenne) 4.5 mg/dl
Pandi Chili (Pimento) 4 mg/dl
160 160 140 120 100 80 60 40 20 0 96 80 70
100 Pepper Capsicum Green chilli Dry chilli Pandichili
Nutritive contents Protein ( Lowry’s Method)
Pepper (Black pepper) 160 mg/dl
Capsicum (Bell pepper) 80 mg/dl
Green Chili (Cayenne) 96 mg/dl
Dry Chili (Cayenne) 100 mg/dl
Pandi Chili (Pimento) 70 mg/dl
100 90 80 70 60 50 40 30 20 10 0 15 9.2
70 Pepper Capsicum Green chilli Dry chilli Pandichili
Nutritive contents Ascorbic acid ( Dichloro-indo-phenol method)
Pepper (Black pepper) 15 mg/dl
Capsicum (Bell pepper) 9.2 mg/dl
Green Chili (Cayenne) 96 mg/dl
Dry Chili (Cayenne) 100 mg/dl
Pandi Chili (Pimento) 70 mg/dl
30 25 20 15 10 5 0
14 12 8
Pepper Capsicum Green chilli Dry chilli Pandichili
Pepper (Black pepper) 28 mg/dl
Capsicum (Bell pepper) 13 mg/dl
Green Chili (Cayenne) 14 mg/dl
Dry Chili (Cayenne) 12 mg/dl
Pandi Chili (Pimento) 8 mg/dl
5 5 4.5 4 3.5 3 2.5 2 1.5 1 0.5 0 1.5 Pepper Capsicum Green chilli Dry chilli Pandichili 3.5 4.5 4
Pepper (Black pepper) 5 mg/dl
Capsicum (Bell pepper) 3.5 mg/dl
Green Chili (Cayenne) 1.5 mg/dl
Dry Chili (Cayenne) 4.5 mg/dl
Pandi Chili (Pimento) 4 mg/dl
0.035 0.03 0.025
Pepper 0.02 0.015 0.015 0.01 0.005 0 0.009 Capsicum Green chilli Dry chilli Pandichili
Pepper (Black pepper) 0.032 mg
Capsicum (Bell pepper) 0.033 mg
Green Chili (Cayenne) 0.30 mg
Dry Chili (Cayenne) 0.009 mg
Pandi Chili (Pimento) 0.015 mg
0.06 0.05 0.04 0.03
0.044 Pepper Capsicum Green chilli Dry chilli Pandichili
0.02 0.01 0
Pepper (Black pepper) 0.056 mg
Capsicum (Bell pepper) 0.056 mg
Green Chili (Cayenne) 0.044 mg
Dry Chili (Cayenne) 0.020 mg
Pandi Chili (Pimento) 0.027 mg
0.002 0.002 0.0018 0.0016 0.0014 0.0012 0.001 0.0008 0.0006 0.0004 0.0002 0 0.00032 0.0004 0.001 0.0008 Pepper Capsicum Green chilli Dry chilli Pandichili
Pepper (Black pepper) 0.001 mg
Capsicum (Bell pepper) 0.00032 mg
Green Chili (Cayenne) 0.0004 mg
Dry Chili (Cayenne) 0.002 mg
Pandi Chili (Pimento) 0.0008 mg
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 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