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Journal of Herbal Medicine and Toxicology 4 (1) 173-177 (2010) ISSN : 0973-4643

Original Article

CHARACTERIZATION OF POLYPHENOLS FROM CORIANDER LEAVES (CORIANDRUM SATIVUM), RED AMARANTHUS (A. PANICULATUS) AND GREEN AMARANTHUS (A. FRUMENTACEUS) USING PAPER CHROMATOGRAPHY: AND THEIR HEALTH IMPLICATIONS

Vanisha S. Nambiar1*, Mammen Daniel2, and Parul Guin1

2

Department of Foods and Nutrition, A WHO collaborating Center for Health Promotion Department of Botany, The Maharaja SayajiRao University of Baroda, Vadodara 390002. Gujarat. India. * Corresponding Author email: [email protected] Received- 15th Oct. 2009, Revised- 10th Nov. 2009, Accepted-23rd Dec 2009

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Abstract : Plant materials, especially GLVs, are recently recommended for prevention of several chronic degenerative disorders (CDDs) due to the antioxidant properties of various flavonoids and phenolic acids present in them. It is therefore important to identify the active phytochemicals present in fruits and vegetables. Scanty data are available on the characterization of polyphenols from vegetables commonly consumed in India. Thus an attempt was made to identify various flavonoids, phenolic acids and glycoflavones present in 3 common Indian green leafy vegetables (GLVs) namely, Coriander leaves (Coriandrum Sativum), Red amaranthus (A. Paniculatus) and Green amaranthus (A. Frumentaceus) leaves using paper chromatography. In Coriander Leaves quercetin, kaempferol and acacetin flavanoids were identified. The phenolic acids identified were vanilic acid, ferulic acid (cis and trans form) and p-coumaric acid. Similar results from red and green amaranth emphasize the role of these GLVs in the prevention of chronic degenerative diseases. Keywords: polyphenols, Antioxidant, Flavonoids, Kaempferol, Polyphenols, Phenolic compounds, Coriander leaves (Coriandrum Sativum), Green amaranthus (A. Frumentaceus), Red amaranthus (A. Paniculatus), Quercetin. INTRODUCTION Presence of phytochemicals in addition to vitamins/ provitamins and minerals, in fruits and vegetables has been recently considered crucial nutritional importance in the prevention of chronic diseases, such as cancer, cardiovascular disease and diabetes[1,2]. A significant inverse correlation has also been reported between total fruits and vegetables intake and cerebrovascular disease mortality. Thus the complex mixture of phytochemicals in fruits and vegetables provide a better protective effect on health than a single phytochemical. Many of these phytochemicals have been found to provide a much stronger antioxidant activity than vitamin C, vitamin E and ß- carotene within the same food. These dietary antioxidants provide bioactive mechanisms to reduce free radical induced oxidative stress. Oxidative stress results from either a decrease of natural cell antioxidant capacity or an increased amount of reactive oxygen species in organisms. When the balance between oxidants and antioxidants in the body is shifted by the over production of free radical, it will lead to oxidative stress and DNA damage [3]. Over the past 10 years, researchers and food manufacturers have become increasingly interested in polyphenols. The chief reason for this interest is the recognition of the antioxidant properties of polyphenols, their great

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abundance in our diet, and their probable role in the prevention of various diseases associated with oxidative stress, such as cancer cardiovascular and neurodegenerative diseases. Our recent study showed that polyphenols, especially, the flavonols such as kaempferol, quercetin and their derivatives are found to be present in leafy vegetables 4 . Flavonoids, as antioxidants, may inhibit the oxidation of LDL cholesterol, reduce platelet aggregation, or reduce ischemic damage. Major flavonoid classes include quercetin, apigenin, luteolin, catechins and soy isoflavones[5-8]. India's flora comprises of 6000 species of plants used for consumption of which around 0.70 metric tons are green leafy vegetables. An inventory on the available green leafy vegetables from the rural, tribal and urban areas of Vadodara and Panchmahal districts of Gujarat, revealed a wide array of plant foods including 37 GLVs, which could be good sources of beta-carotene [9-10]. Twenty-five of these GLVs were used for medicinal purposes by the local population. These health promoting properties along with the fact that several of these greens may be potential sources of beta carotene along with other micronutrients make these GLVs an important nominee for their use in the food based approach to combat vitamin A deficiency[11-18]. Though information is available on the nutrient profile of most of these, there is no data available on their polyphenols content. The literature search revealed data on GLVs familiar to the western world such as broccoli, lettuce, spinach, green tea. Consequently there is a need to generate data about the polyphenol profile of Indian GLVs so as to bridge the gap between the known and the unknown. The present paper highlights the results of the polyphenol composition of three GLVs commonly consumed in India, namely, Coriander leaves (Coriandrum Sativum), Red amaranthus (A. Paniculatus) and Green amaranthus (A. Frumentaceus), which may be a repository of several favourable chemicals. METHODS AND MATERIALS In the present study 3 green leafy vegetables were taken for the separation and identification of polyphenols. They were:(A) Coriander leaves (Coriandrum Sativum) (B) Red amaranthus (A. Paniculatus) (C) Green amaranthus (A. Frumentaceus). Five kg of each type of leaves were obtained from 3 different vegetable markets of Vadodara city, mixed, cleaned, shade dried and powdered using a laboratory blender, stored under refrigeration and used in triplicates for the identification of the polyphenols by paper chromatpgraphy as described in our earlier paper[4]. The Polyphenol separation included the isolation and identification of:(a) Flavonoids (b) Phenolic Acids (c) Glycoflavones The standard analytical procedures involving interaction with diagnostic reagents and paper chromatographic separation of compounds and their UV/Visible spectroscopic studies including hypsochromic and bathochromic shifts with reagents such as AlCl3, AlCl 3/HCl, NaOMe, NaOAc and NaOAc/H3PO3 were followed for the identification of flavonoids and other phenolics. The identities of all the compounds were confirmed by co-chromatogaphy (paper and thin-layer chromatography) with authentic samples[19-21]. RESULTS AND DISCUSSION In Coriander Leaves the flavonoids that were identified are quercetin, kaempferol and acacetin. The phenolic acids that were identified vanilic acid, ferulic acid (cis and trans form) and p-coumaric acid as can be seen from Table 1. No glycoflavones were detected. No literature is available regarding the polyphenol composition of coriander leaves, red or green amaranth by other investigators. Quercetin is found in abundance in onions, apples, broccoli and berries. Thus coriander leaves, rich in quercetin, can be an important food source for the prevention of chronic degenerative diseases. The flavonol quercetin (3, 3', 4', 5, 7- pentahydroxyflavone) is one of the most abundant dietary flavonoids. Quercetin and other flavonoids have been shown to modify eicosanoid biosynthesis (antiprostanoid and anti-inflammatory responses), protect low density lipoprotein (LDL) from oxidation (prevention of atherosclerotic plaque formation) and promote relaxation of cardiovascular

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Table 1 :

Polyphenol content of three Green leafy vegetables

Coriander leaves + + + + + Green amaranthus leaves Red amaranthus leaves -

Name of the compounds Apigenin 3',4'-di-OMe luteolin Kaempferol 4'-OMe kaempferol 7'4'-di-OMe kaempferol Quercetin 3'-OMe quercetin 4'-OMe quercetin 3',4'-di-OMe quercetin Acacetin Gossypetin Quercetagetin Proanthocyanidins Anthocyanins Coumarins + : Present; - : Absent

Table 2 :

Phenolic acids content of three Green leafy vegetables

Coriander leaves + + + + Green amaranthus leaves + + + + + Red amaranthus leaves + + + +

Name of the compounds Vanillic acid Syringic acid p-OH benzoic acid Melilotic acid Gentisic acid o-Coumaric acid p-Coumaric acid Cis-Ferulic acid Trans-Ferulic acid Phloretic acid Chlorogenic acid Resorcylic acid + : Present ; - : Absent

smooth muscle (antihypertensive, antiarrythemic effects). In addition, flavonoids have been shown to have antiviral and anticarcinogenic properties[22].

Several flavonoids, including quercetin results in a reduction in the ischemia-reperfusion injury by interfering with inducible nitric-oxide synthase activity.

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Nitric oxide reacts with free radicals, thereby producing peroxynitrite can directly oxidize LDL, resulting in irreversible damage to the cell membrane. When flavonoids are used as antioxidants, free radicals are scavenged and, therefore can no longer react with nitric oxide, resulting in less damage. Coriander is also reported to be a chelating agent and reported to be effective as pharmaceutical agents in removing heavy metals[23]. Results of the Green Amaranthus reveled no presence of either flavonoids or glycoflavones. However several phenolic acids were identified, namely: vanillic acid, cis and trans ferulic acid, p- OH benzoic acid, ocoumaric acid, p- coumaric acid and melilotic acid as can be seen from Table 1. In Red Amaranths also no flavonoids or glycoflavones were detected. The phenolic acids that were identified were vanilic acid, p- OH benzoic acid, p-coumaric acid and syringic acid. Two classes of phenolic acids can be distinguished: derivatives of benzoic acid and derivatives of cinnamic acids. The hydroxybenzoic acid content of edible plants is generally very low, with the exception of certain red fruits, black radish and onions. The hydroxylcinnamic acids are more common than the hydroxybenzoic acids and consist chiefly of pcoumaric, caffeic, ferulic and sinapic acids. These acids are rarely found in the free form, except in processed food that has undergone freezing, sterilization or fermentation. Phenolic compound ferulic acid has added health benefits as it battles cancer. Ferulic acid is the predominant bound phenolic form. The fact that ferulic acid could be identified in all the three GLVs, emphasizes their potential role in the fight against cancer[24-29]. These results emphasize that food-based approaches are more practical and sustainable to combat various health or nutritional disorders. Inclusion of these greens in the daily diet will not only add on to the nutritional value of the diet, but also serve as an additional advantage of being functional foods, which would maintain heart health due to their high antioxidant activity. These green could also be teamed up with several other functional foods such as foods containing plant sterols and stanols, fatty fish, foods containing fiber, nuts, and flaxseed, garlic, tea, grapes, dark chocolates and many more. Inclusion of these foods along with healthy dietary patterns and lifestyle can help fight against obesity, diabetes and heart disease. Further studies are suggested to quantify these polyphenols and assess their health benefits by conducting clinical trials. ACKOWLEDGEMENT We thank all the staff and students of the phytochemistry laboratory, Department of Botany for their cooperation and support towards completion of this study. REFERENCES

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