Phytochemical profiling of Garcinia gummi-gutta (Malabar tamarind) and in vitro analysis of cholesterol lowering effect

Table of contents

Table of figures

Table of tables

List of abbreviations

Phytochemical profiling of Garcinia gummi-gutta (Malabar tamarind) and in vitro analysis of cholesterol lowering effect

Abstract

1. Introduction
1.1 Objectives

2. Review of literature

3. Hypothesis

4. Materials and Methods
4.1 Study area
4.2 Collection of plant material
4.3 Preparation of Garcinia gummi-gutta fruit pulp extracts
4.4 Phytochemical screening
4.5 Preparation of cholesterol samples
4.7 Estimation of cholesterol
4.8 Statistical analysis

5. Results and discussion

6. Conclusions

Acknowledgements

References

ACKNOWLEDGEMENTS

Firstly we thank God Almighty whose blessing were always with us and helped us to complete this project work successfully.

We wish to thank our beloved Manager Rev. Fr. Dr. George Njarakunnel, Respected Principal Dr. Joseph V.J, Vice Principal Fr. Joseph Allencheril, Bursar Shaji Augustine and the Management for providing all the necessary facilities in carrying out the study. We express our sincere thanks to Mr. Binoy A Mulanthra (lab in charge, Department of Biotechnology) for the support. This research work will not be possible with the co-operation of many farmers.

Lastly, we extend our indebt thanks to patents, friends, and well wishers for their love and support.

Prem Jose Vazhacharickal*, Jiby John Mathew, Sajeshkumar N.K, Ratheesh Mohanan and Preeja Mol MR

*Address for correspondence

Assistant Professor

Department of Biotechnology

Mar Augusthinose College

Ramapuram-686576

Kerala, India

Table of figures

Figure 1. Map of Kerala showing the soil sample collection point. Authors own work

Figure 2. Details of Garcinia gummi-gutta (Malabar tamarind) plant. Photo courtesy: Wikipedia

Figure 3. Details of phytochemical analysis of Garcinia gummi-gutta fruit (methanol extract); A.Proteins, B.Alkaloids, C.Saponins, D.Steroids, E.Coumarins, F.Molischs, G.Flavanoids, H. Phlobatannins, I.Leucoanthocyanines, J.Anthocyanines, K.Phenols, L.Emodines, M.Anthraquinones, N.Glycosides, O. Terpenoids. Authors own image

Figure 4. Details of phytochemical analysis of Garcinia gummi-gutta fruit (water extract); A.Proteins, B.Alkaloids, C.Saponins, D.Steroids, E.Coumarins, F.Molischs, G.Flavanoids, H. Phlobatannins, I.Leucoanthocyanines, J.Anthocyanines, K.Phenols, L.Emodines, M.Anthraquinones, N.Glycosides, O. Terpenoids. Authors own image

Figure 5. Standard graph for cholesterol estimation by Zak’s method

Table of tables

Table 1. Nutritional value of Garcinia gummi-gutta fruit extracts

Table 2. Preliminary phytochemical analysis of Garcinia gummi-gutta fruit extracts

Table 3. Cholesterol estimation at different time intervals after treatment (n=3; values in mg/g sample)

List of abbreviations

Abbildung in dieser Leseprobe nicht enthalten

Phytochemical profiling of Garcinia gummi-gutta (Malabar tamarind) and in vitro analysis of cholesterol lowering effect

Prem Jose Vazhacharickal1*, Jiby John Mathew1, Sajeshkumar N.K1 , Ratheesh Mohanan2 and Preeja Mol MR1

* premjosev@gmail.com

1 Department of Biotechnology, Mar Augusthinose College, Ramapuram, Kerala, India-686576

2 Department of Biochemistry, St. Thomas College, Pala, Kerala, India-686574

Abstract

In our society cholesterol related diseases are increasing day by day. The major reason for this is our eating habit or food habit. Eating foods containing trans fats, which are often found in fast food and commercially baked breads, cookies, and snack foods. It was observed that while peoples taking the fruits of Garcinia gummi-gutta in their food preparations have low cholesterol level. Some peoples are eating boiled fruits and drinking water in which the fruits peels are boiled in order to reduce the cholesterol level. This study was carried out to explore the phytochemical constituents of the solvent extracts of Garcinia gummi-gutta fruit and in vitro analysis of its cholesterol lowering effect. The cholesterol lowering effect was carried out in vitro using common fatty food materials like ghee, fats of pork and chicken, hen egg and cod liver oil. Each food materials were treated with extract and incubated for a number of days and each day the cholesterol level was estimated by Zak’s method. From the data, pork and chicken fat, egg yolk and ghee shows significant reduction in the cholesterol level. From the present study it can be concluded that the constituents present in the extracts may be responsible for the cholesterol lowering activity.

Keywords: Cholesterol; Zak’s method; Hypercholesterolemia, Malabar tamarind, Emodins, Coumarins.

1. Introduction

Plants are the gifts of nature to human beings for basic preventive and curative healthcare since time immemorial. Plant extract contains various secondary metabolites that have been reported scientifically for biological activities and can also protect humans against infectious diseases (Jeeva et al., 2006; Balakumar et al., 2011). These metabolites include active ingredients like alkaloids, flavonoids, steroids, glycosides, terpenes and tannins (Sukumaran et al., 2012). Due to wide range of biological activity these phytoconstituents can be used for the treatment of various diseases (Dahanukar et al., 2000; Iwu et al., 1993). To promote the proper use and to determine their potential as sources for new drugs, it is essential to study the medicinal plants (Parekh et al., 2007). In India almost 95% of the traditional systems like Unani, Ayurveda, Siddha and Homeopathy have plant based prescriptions (Satyavati et al., 1976). According to world health organization (WHO) about 80% of population in developing countries relies on traditional medicines for their primary health care needs (Jayer et al., 2007).

Numerous phytoconstituents have been found to be active against many degenerative diseases and pathological processes in humans such as in ageing, coronary heart disease, Alzheimer’s disease, neurodegenerative disorders, atherosclerosis cataracts and inflammation (Birt, 2006). The Genus Garcinia contains around 200 species, where 35 species are found in the two ecosystems of India, the Western Ghats and the Himalayan foot hills (Parthsarathy et al., 2014). It is a hardwood underutilized medicinal fruit crop, where its fruits are used in food preparations, especially in curries (Heber, 1982). Previous studies had shown that, the leaf extract of the plant had shown increase the levels of erythrocytes, leucocytes, thrombocytes, hemoglobin and was found to decrease the level of glucose, total cholesterol and low density lipoprotein (LDL) level in catfish. This was due to the presence of a hypolipidaemic compound, hydroxy citric acid. It also increased immunity by increasing cellular immunological indicators (Prasad et al., 2007; Tian et al, 2009).

At present peoples are consuming more fatty food like meat and chicken, dairy products and palm and coconut oil. This fatty food product contains saturated fat. It can be found in processed foods like biscuits, pastries and takeaway foods that have used ingredients like butter, palm oil, cheese and meat. The over consumption of saturated fat food increases the LDL level in the blood. To reducing the LDL level peoples are using more ‘kudampuli’ (Garcinia gummi-gutta) in their food preparations. They believe that some chemical present in the Garcinia gummi-gutta fruits will lower cholesterol level. Hence this study was carried out to explore the phytochemical constituents of the solvent extracts of Garcinia gummi-gutta fruits and its cholesterol lowering efficiency.

1.1 Objectives

The objectives of this study to evaluate the phytochemical properties of methanol and water extract of Garcinia gummi-gutta fruit extracts and its cholesterol lowering effect on various fatty food materials.

2. Review of literature

Medicinal plants, medicinal herbs, or simply herbs have been identified and used from prehistoric times. Plants make many chemical compounds for biological functions, including defence against insects, fungi and herbivorous mammals. Over 12,000 active compounds are known to science. These chemicals work on the human body in exactly the same way as pharmaceutical drugs, so herbal medicines can be beneficial and have harmful side effects just like conventional drugs. However, since a single plant may contain many substances, the effects of taking a plant as medicine can be complex.

Many plants produce chemical compounds for defence against herbivores. These are often useful as drugs (Springbob et al., 2009; Elumalai and Eswariah, 2012).

Plant medicines are ubiquitous in pre-industrial societies, while some 7,000 conventional medicines such as aspirin, digitalis, opium, and quinine derive directly from traditional plant medicines, accounting for around a quarter of the modern pharmacopoeia. They are in general far cheaper, and many can be home-grown or picked for free. Further, pharmaceutical companies have made use of the herbal knowledge of indigenous people around the world to search for new drug candidates (Duke, 2000; Kala and Sajwan, 2007). In India, where Ayurveda has been practiced for centuries, herbal remedies are the responsibility of a government department, AYUSH, under the Ministry of Health & Family Welfare. Traditional Chinese medicine makes use of a wide variety of plants, among other materials and techniques.

Plant medicines including opiates, cocaine and cannabis have both medical and recreational uses. Different countries have at various times made some uses of drugs illegal, partly on the basis of the risks involved in taking psychoactive drugs.

Cholesterol is both our friend and foe. At normal levels, it is an essential substance for the body. However, if concentrations in the blood get too high, it becomes a silent danger that puts us at risk of a heart attack.

Cholesterol is found in every cell of the body and has important natural functions when it comes to digesting foods, producing hormones, and generating Vitamin D. It is manufactured by the body but can also be taken in from food. It is waxy and fat-like in appearance.

Cholesterol is an oil-based substance and does not mix with the blood, which is water-based. It is carried around the body through the blood by lipoproteins.

Two types of lipoprotein carry the parcels of cholesterol:

Low-density lipoprotein (LDL) - cholesterol carried by this type is known as 'bad' cholesterol.

High-density lipoprotein (HDL) - cholesterol carried by this type is known as 'good' cholesterol.

Contributing to the structure of cell walls, making up digestive bile acids in the intestine, allowing the body to produce vitamin D, and enabling the body to make certain hormones.

High cholesterol is a major risk factor for coronary heart diseases, a cause of heart attacks. Reducing blood lipid levels may lower cardiovascular risk. Elevated levels of LDL lead to a build-up of cholesterol in the arteries, whereas HDL carries cholesterol to the liver for removal from the body. A build-up of cholesterol is part of the process that narrows arteries, called atherosclerosis, in which plaques form and cause restriction of blood flow.

Limiting intake of fat in the diet helps manage cholesterol levels .Cholesterol (from animal foods, meat, and cheese). Saturated fat (found in some meats, dairy products, chocolate, baked goods and deep-fried and processed foods) Tran’s fats (found in some fried and processed foods)

Being overweight or obese can also lead to higher blood LDL levels. Regular exercise may help manage this risk factor. The primary causes of high cholesterol are genetic - very high LDL levels are found in the inherited condition familial hypercholesterolemia.

Diabetes, liver or kidney disease, polycystic ovary syndrome, pregnancy and other conditions that increase levels of female hormones, underactive thyroid gland, drugs that increase LDL cholesterol and decrease HDL cholesterol (progestin’s, anabolic steroids, and corticosteroids)

Having high cholesterol levels, while a risk factor for other conditions, does not itself present any signs or symptoms. Unless routinely screened through regular blood testing, high cholesterol levels will go unnoticed and could present a silent threat of heart attack or stroke.

It is a sterol (or modified steroids), a type of lipids molecule, and is biosynthesized by all animal cells, because it is an essential structural component of all animal cell membranes; essential to maintain both membrane structural integrity and fluidity. Cholesterol enables animal cells to dispense with a cell wall (to protect membrane integrity and cell viability), thereby allowing animal cells to change shape rapidly and animals to move.

In addition to its importance for animal cell structure, cholesterol also serves as a precursor for the biosynthesis of steroid hormones, bile acid (Hanukoglu, 1992) and vitamin D. Cholesterol is the principal sterol synthesized by all animals. In vertebrates, hepatic cells typically produce the greatest amounts. It is absent among prokaryotes (Archae and bacteria), although there are some exceptions, such as mycoplasma, which require cholesterol for growth (Razin and Tully, 1970).

This review is emphasizes on traditionally used clinically potential plant Garcinia gummi-gutta. The plant Garcinia gummi-gutta {L.} Robson {G.cambogia, G.quaesita} belongs to the family Guttiferae {Clusiferae}. It is a wild tropical and subtropical plant. These polygamous trees or shrubs are distributed widely throughout the tropical Asia, Polynesia and African countries and have tremendous potential as spice and medicinal plants (Soni et al., 2004). The genus Garcinia includes 390 species, of which about 30 different species are reported to be grown in Andaman islands, north east hill region, west Bengal, Orissa, Bihar, Western Ghats covering Maharashtra, Goa, Karnataka, Kerala and Nilgiri hills in India (Krishnamurthy et.al, 2006; Parthasarathy and Karthikeyan, 1997; Ramesh and Pascal, 1997). The commonly used tamarind pant is said to be indigenous to Africa and was introduced to India (Thas 1979). Garcinia indica, Garcinia gummi-gutta, Garcinia xanthochymus and Garcinia spicata are widely distributed in the Western Ghats, of which the former two are the most wildly spread species in Goa region of Konkan belt and northern part of Kerala respectively, with tremendous natural variability. Garcinia gummi-gutta has a limited native global distribution, being restricted to India, Nepal and Sri Lanka but it has been introduced elsewhere where it is distributed in the subtropical region of Asia including China, Malaysia and the Philippines. These trees are found mainly in the semi evergreen to evergreen forests of southwest India, predominantly in the Western Ghats (Maharashtra, Karnataka, Kerala, and Tamil Nadu). This tree can grow on both hilltops and plain lands but grows best in dry or occasionally waterlogged or flooded soils in riverbanks and valleys. It is tolerant to fluctuating water tables and drought (Abraham et al., 2006; Orwa et al., 2014).

Garcinia gummi-gutta is an evergreen, small or medium- sized dioeciously under storey tree, 5-20 m tall, with a rounded crown and horizontal or drooping branch. Young branch lets are substerete and glabrous and the trunk and bark is reddish brown and lenticellate. The leaves are dark green, shiny, opposite and decussate with 5-16 cm long petioles and 5-13 cm laminate. The leaves are elliptic, oblanceolate to obovate in shape, and the apex is usually acute and rarely obtuse. The polygamous flowers are in axillaries or terminal clusters and the sepals are cream while the petals are pink, red in colour. The flowers in clusters of 4-20. Petals normally 4, each about 12 mm wide long; anthers attached to a pistil lode with a non functional stigma. Female flowers occur singly or in clusters of up to 4. The stigmatic surface is normally enlarged and there is no style. Pistil late flowers have rudimentary and non-functional staminodes. Neither male nor female flowers produce nectar. Flowering occurs during the summer (March-May) while fruiting occurs during the rainy season (June- September). The ovoid fruits are about 5 cm in diameter with 6 to 8 grooves. The fruit can be yellow, orange or red when ripe and has 6 to 8 seed surrounded by a succulent aril (Martin et al., 1987). Growth of the tree is slow and differentiation between male and female trees is known only at the flowering stage, at approximately 7- 9 years (Kalia et al., 2012).

Seed grown plants start bearing after 10- 12 years whereas grafts from the third year onwards and will attain the stage of full bearing at the age of 12 -15 years. In India, flowering occurs in January- march and fruits mature in July. There are also reports of off-season bearers, bearing twice annually. The orange yellow mature fruits either drop from the tree or are harvested manually. The rind is separated for processing immediately after harvested (Chacko and Pillai, 1997).

Garcinia gummi-gutta flowers in the dry season. It appears to be pollinated by wind, bees and small weevils of the genus Deleromus (Curculionidae). Monkeys (Presbytus entellus and Macacaradiate) and species of civets (Paradoxorus hermaphrodites and Paradoxorus jerdonii) disperse the fruits. The seeds are consumed by two species of arboreal squirrels (Ratufa indica and Funambulus palmaram) (Chacko and Pillai, 1997).

Garcinia cambogia is an economically important spice tree valued for the sun- dried smoked rind which is widely used as a flavour condiment, especially in fish curries. The dried rind of the fruit acts as a bacteriostatic agent and in combination with salt is used to cure fish in India and Sri Lanka (Anilkumar et al., 2002; Jena et al., 2002). It is also used as a substitute of kokum butter and as a common additive to make meals more filling (Burdock et al., 2005). The fruit rind is used medicinally to treat rheumatism and bowel complaints and it is employed as a purgative, hydrogogue, anthelmintic and emetic. It is also used in veterinary medicine where a rinse is used to treat mouth diseases in cattle (Abraham et al., 2006). The fruits are edible, but very acidic and are not generally eaten raw (Jena et al., 2002). A tonic prepared from the fruit, which contains high concentrations of vitamin c, Is used in India to treat various heart diseases (Pandey et al., 2013). Garcinia gummi-gutta is not only used medicinally; the rind is used to polish gold and silver ornaments and as a substitutes for acetic acid for the coagulation of rubber latex; the gum is used as a varnish and the resin is used as a pigment in miniature paintings and water colours (Abraham et al., 2006).

Nutritional Values of Garcinia gummi-gutta

The rind of Garcinia gummi-gutta comprises of tannin (1.7%), pectin (0.9%), fat (1.4%), moisture (80.0 g/100 g), protein (1%) and sugars (4.1%). The seed is an excellent source of stearic triglycerides, stearic and oleic acid. The leaves contain fibre (1.24 g), carbohydrates (17.2 g), moisture (75%), calcium (250 mg), protein (2.3 g), ascorbic acid (10 mg), fat (0.5 g), iron (15.14 mg), and oxalic acid (18.10 mg). Also, the plant contains a minimal amount of citric and hydroxyl citric acid lactones.

Table 1. Nutritional value of Garcinia gummi-gutta fruit extracts.

Abbildung in dieser Leseprobe nicht enthalten

Common names are Bitter kola, brindali berries, brindal berry, garcinia kola, Malabar tamarind, Garcinia gummi-gutta.

In Ayurveda, it is said that the sour flavours, such as those from Garcinia, activate digestion. Garcinia has also been considered to make foods more filling and satisfying, and has been used routinely for many centuries with no known toxicity. This herb has been used historically in India to support the treatment of various health conditions. The yellow resin obtained from the fruit is soluble in turpentine and used as varnish.

Apart from the traditional uses of Garcinia cambogia, it finds a wide value of applications in "Neutraceutical" field. Neutraceutical is the word used for any nutritional 1 herbal product which is marketed as OTC (over the counter sale) products. The major market is in US, followed by Japan and Europe. Garcinia cambogia is a revolutionary component in neutraceutical 1 dietary supplement areas as a source of hydroxy citric acid (HCA), which is known as a weight reducing agent (Sullivan and Triscari, 1977). In addition to tablets and capsules, it is marketed as biscuits, chewing gum, snack bar etc. Water-soluble HCA is available as soft drinks and beverages the modern applications are based on the fact that in the body carbohydrates of the food are broken down into glucose which is stored as glycogen. When glycogen storage is saturated, excess glucose is converted into fat and cholesterol. Garcinia gummi-gutta extract inhibit body's conversion of glucose into fat and cholesterol by inhibiting certain enzyme process (Sullivan et al., 1973). The increase in glycogen stores, help significantly reduce cravings for food, reduce appetite and induce weight loss.

It is a hardwood underutilized medicinal fruit crop, where its fruits are used in food preparations, especially in curries (Nair et al., 2005; Heber, 1873). Previous studies had shown that, the leaf extract of the plant had shown strong antifungal activity on three fungal species namely Phytopthora sp., Curvularia sp. and Corynesporia sp., which could be used as natural fungicide (Dhanya and Benny, 2013). Garcinia gummi-gutta had also shown to increase the levels of erythrocytes, leucocytes, thrombocytes, hemoglobin and was found to decrease the level of glucose, total cholesterol and LDL level in catfish. This was due to the presence of a Hypolipidaemic compound, hydroxy citric acid. It also increased immunity by increasing cellular immunological indicators (Prasad and Priyanka, 2011; Tian et al., 2009; Olila and Opuda-Asibo, 2001; Nikhat et al., 2009).

The bioactive molecules like HCA, flavonoids, terpenes, polysaccharides, procyanidines and polyisoprenylatedbenzophenone derivatives like garcinol, xanthochymol and guttiferone were isolated from the genus Garcinia. It was found that the polyisoprenylatedbenzophenone and xanthone derivatives have antioxidant, apoptotic, anti-cancer, anti-, antibacterial, anti-viral, anti-fungal, anti-ulcer and anti-protozoan properties (Naveen and Krishnakumar, 2013). The long term medication of antibiotics such as tetracycline, erythromycin etc., and over usage of synthetic antioxidants like Propylgallate etc., has a possibility for causing health hazards and side effects (Tian et al., 2009). The main aim of this study was to find a natural therapeutic compound which has antibacterial property and antioxidant property so that it can be an alternative in treating drug resistant bacteria.

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