Middle-East Journal of Scientific Research 26 (6): 621-629, 2018 ISSN 1990-9233 © IDOSI Publications, 2018 DOI: 10.5829/idosi.mejsr.2018.621.629

Essential and Trace Elements Analysis of ganitrus Fruit by Wavelength Dispersive X-Ray Fluorescence Spectrometer and its Impact on Health

1,2Sunil Jawla, 3 Ramesh Sharma and 4 D.V. Rai

1Atam Institute of Pharmacy, Garhi, Hansi, Hisar, NH 9, Haryana, India 125033. 2Adarsh Vijendra Institute of Pharmaceutical Sciences, Shobhit University, Gangoh, Saharanpur, UP, India 247341 3Sophisticated Analytical Instrumentation Facility, Panjab University, Chandigarh, India 160014 4Centre for Biological Engineering, Shobhit University, Gangoh, Saharanpur, UP, India 247341

Abstract: Elaeocarpus ganitrus (Rudraksh) is an important traditional medicinal in Indian sub-continent. The medicinal and healing properties of this plant and its fruit are documented in ancient literature reports. As per these reports and consumption of fruit by animals and birds indicates the nutritional importance of this plant. Elemental analysis of pulp and bead of fruit were carried out by using Wavelength dispersive X-ray fluorescence spectrometer. Various essential and trace elements were detected in fruits, which is having nutritional importance and having impact on human health. The elemental concentrations (mg/100 g) were also significantly (P < 0.05) varied and ranged in fruit pulp and bead. The major minerals like calcium, magnesium, phosphorus, potassium, chlorine, sodium and sulphur were detected in both bead and pulp. The pulp was found to be very rich in potassium (1470 mg / 100 g), followed by calcium (240 mg / 100 g) in bead. Trace elements iron, copper, zinc, molybdenum, palladium, rubidium and silicon were detected in both bead and pulp. However manganese, aluminium, strontium, nickel and selenium were detected in traces in beads, indicating bead are richer in trace elements, which supports its ethno-medicinal importance and nutritional value. These trace elements are essential prosthetic group of many metaloenzymes involved in body’s metabolism and various cellular processes. Hence increase in production and consumption of nutrient rich Rudraksh fruit may contribute to supplement/formulate the diets and alleviate the risk associated with malnutrition in the developing countries.

Key words: Essential and Trace Elements Fruit Pulp and Bead Elaeocarpus ganitrus Rudraksh Nutritional Value Wavelength Dispersive X-Ray Fluorescence Spectrometer

INTRODUCTION tissues is essential for preserving health. Micronutrients play a significant role in various regenerative processes Essential elements are vital for most of the metabolic and enhance immunity of the body against pathogens and physiologic processes of human body. The trace [1, 2]. elements are required in small quantities but play an Traditionally fruits remain a valuable sources of important role in maintaining various biological processes. essential and trace elements in addition to their medicinal They are part of metalloenzymes mediating vast number properties. Food and nutritional insecurity among adults of metabolic processes. Excess amount of these trace and children remains a challenge in many developing elements may be toxic and appropriate nutrition of cells & countries. family is distributed in tropical

Corresponding Author: Sunil Jawla, Atam Institute of Pharmacy, Garhi, Hansi, Hisar, Haryana, India 125033. Adarsh Vijendra Institute of Pharmaceutical Sciences, Shobhit University, Gangoh, Saharanpur, UP, India 247341.

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beads were dried at room temp under shade and powdered. Essential and trace elemental analysis of pulp and bead powder were carried out [23, 24].

Mineral Analysis: In present study, the S8 TIGER wavelength dispersive X-ray fluorescence spectrometer (WDXRF) (Bruker, Germany) available at SAIF Department of Panjab University, Chandigarh, India was used for elemental analysis. The pulp and bead powder (Elaeocarpus ganitrus) was pressed using a pellet making machine with a pressure of 40 tons within 20s to form a Fig. 1: Elaeocarpus ganitrus Blume Tree Bearing Fruits stable 8gm pellet (3mm thickness and 35 mm diameter). The quantitative analysis was performed by inbuilt and subtropical region of the world. Around 25 species software with WDXRF system SPECTRA PLUS in are distributed around India, including Bangladesh, triplicate. The concentration of essential elements sodium, Bhutan, Nepal, Pakistan and Sri Lanka [3, 4]. The fruiting potassium, chloride, calcium, phosphorous, magnesium period of Elaeocarpus ganitrus is in the month of and sulphur were determined in bead and pulp powder of November and December (Fig. 1). Elaeocarpus ganitrus (Table 1) [25]. The plant is used as medicines due to The pulp was very rich in potassium (1470 mg / 100 pharmacological properties by local people for cure of g), the most abundant macro-element in Elaeocarpus various ailments as reported in literature. The are ganitrus pulp, followed by calcium (240 mg / 100 g) in explored continuously in search of new lead compounds bead and chlorine (200mg / 100 g) in pulp. The other [5-8]. In Indian subcontinent, Rudraksh (Elaeocarpus elements detected in descending order are magnesium, ganitrus; syn. E. sphaericus) is known as a holy tree as sodium, manganese, zinc, iron and copper. reported in Ayurvedic literature and dedicated to Lord Trace elements iron, copper, zinc, molybdenum, Shiva. The pharmacological properties like anti- palladium, rubidium and silicon were detected in both convulasnt, antihypertension, anti-inflammatory and bead and pulp. Chromium, bromine and arsenic were antimicrobial activity of Rudraksh are reported by various found in traces in fruit pulp. However manganese, researchers [9-14]. Research on different prospective of aluminium, strontium, nickel and selenium were detected Rudraksh plant is in progress [15, 16]. Indolizidine in traces in beads (Table II). alkaloids isolated from Elaeocarpus family, are considered as potential chemotherapeutic agents for cancer, diabetes, Data Analysis: The statistical analysis of elemental viral infections and immunosuppressive deficiencies composition of Elaeocarpus ganitrus fruit pulp and bead [17-22]. Consumption of fruit by animals and birds as were carried out to draw the significance information. reported in literature, indicates the nutritional importance The data analysis was carried out according to a of this plant. Therefore, the aim of present work to completely randomized design, representing the mean evaluate the nutritional importance of Elaeocarpus values of triplicate analysis or more, depending on the ganitrus fruit pulp and bead. availability of the data [26].

MATERIALS AND METHODS RESULTS AND DISCUSSION

Sample Preparation: Elaeocarpus ganitrus Blume fresh Minerals are vital for the mental & physical health fruits were collected from Dehradun, India, in February, and are main constituents of bones, teeth, muscles, blood, 2017. The plant material was authenticated from Botanical tissues and nerve cells. They help in maintenance of acid Survey of India, Northern Regional Centre, Dehradun, base balance, response of nerves to physiological India (Voucher specimen no. 115875). The pulp was stimulation and also blood coagulation [27]. There are removed manually from the fruit and dried under the two types of minerals needed to stay healthy: major shade at room temperature and lyophilised. Similarly or macrominerals and trace minerals. Body needs large

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Table 1: Macro-nutrients Analysis of Elaeocarpus ganitrus Fruit Pulp and (240 mg / 100 g) in bead. The details of macro nutrients Bead estimated in Elaeocarpus ganitrus fruit pulp and bead a Content (mg/100g) along with their biological importance are as follows. ------Mineral Pulp Bead Potassium: Potassium is a very significant body mineral, Macro Potassium 1470±0.127 110±0.223 Calcium 50±0.234 240±0.531 play a vital role in cellular and electrical function. Phosphorus 130±0.011 20±0.278 Potassium is the primary positive ion found within the Magnesium 80±0.075 60±0.342 cells other than sodium and chloride. The blood serum Sodium - 10±0.057 contains about 4-5 mg/100ml of the total potassium Sulphur 110±0.861 40±0.451 whereas RBC contain 420 mg. Potassium is useful in iron Chlorine 200±0.762 30±0.213 utilization [30] to control hypertension in patients on a Means values±SD of three determination diuretics and suffering from excessive excretion of

Table 2: Micro-nutrients Analysis of Elaeocarpus ganitrus Fruit Pulp and potassium [31]. The potassium content of Rudraksh fruit Bead pulp and bead was found to be 1470 & 110 mg/100 Contenta (mg/100g) respectively indicating the importance of fruit in ------potassium nutrition. Potassium level is more in pulp as Mineral Pulp Bead compared to bead and may be main source of potassium Micro Iron 0.082±0.361 80±0.351 (Table 1). Copper 0.014±0.251 0.012±0.246 Zinc 0.012±0.045 0.018±0.119 Calcium: Calcium is the major mineral found in bones, Silicon 30±0.276 70±0.029 teeth and blood. Daily need of adult is about 1 g of Manganese - 11±0.053 Aluminium - 20±0.048 calcium per day. It is necessary for blood coagulation and Palladium 0.015±0.251 0.026±0.072 for intracellular cement [32]. The calcium content of Rubidium 0.023±0.451 0.026±0.028 Rudraksh fruit pulp and bead was found to be 50 & Molybdenum 0.014±0.052 0.016±0.059 240 mg/100 respectively indicating the importance of fruit Strontium - 0.011±0.117 in calcium nutrition. Bead contains more level of calcium Nickel - 0.007±0.341 as compared to pulp and may be a potential candidate for Selenium - 0.006±0.043 calcium supplement (Table 1). Chromium 0.009±0.122 - Bromine 0.006±0.041 - Arsenic 0.004±0.035 - Phosphorous: Phosphorous is the second most abundant aMeans values ± SD of three determination mineral in the human body, representing about 1% of total body weight. Due to its extracellular and intracellular amounts of major minerals and only very small amounts of distribution, phosphorus functions as a structural trace minerals for normal function. Major minerals are component of bones & teeth, DNA/RNA, enables the calcium, magnesium, phosphorus, potassium, chloride and bipolarity of lipid membranes and circulating lipoproteins sulphur (Table 1). [29]. Phosphorus has important role in production and Trace minerals are iron, copper, zinc, manganese, storage of energy in phosphate bonds (ATP), blood fluoride, selenium and cobalt (Table 2). Both major and buffer, gene transcription regulation, enzyme catalysis trace minerals are vital for body functions and processes. and signal transduction of various regulatory pathways. Without minerals body is not able to build new tissue, flex Extracellular space contains less than 1% of inorganic and contract muscles, transmit nerve impulses, clot blood, phosphate which is metabolically active and serum maintain a neutral pH and keep heart beating [28, 29]. concentration 2.5–4.5 mg/dL in adults [33]. The phosphorous content of Rudraksh fruit pulp and bead Macro-Nutrients: The concentration of essential was found to be 130 & 23 mg/100 g respectively indicating elements sodium, potassium, chloride, calcium, their nutritional importance (Table I). phosphorous, magnesium and sulphur were determined in bead and pulp powder of Elaeocarpus ganitrus by using Chlorine: Chlorine is a component of body secretions wavelength dispersive X-ray fluorescence spectrometer resulting from anabolism and catabolism. It is store to (Table 1). The most abundant macro-element in fruit pulp limited extent in the skin, subcutaneous tissues & was potassium (1470 mg / 100 g), followed by calcium skeleton and about 66% as anion in the blood. It is

623 Middle-East J. Sci. Res., 26 (6): 621-629, 2018 essential for the electrical neutrality of extracellular fluids Micro-Nutrients: Trace elements iron, copper, zinc, and help in maintenance of acid-base balance. Gastric molybdenum, palladium, rubidium and silicon were juices composed of chlorides in the form of hydrochloric detected in both bead and pulp. Chromium, bromine and acid. The level of chlorides may become dangerously low arsenic were found in traces in fruit pulp. However during dehydration, severe vomiting and diarrhoea and in manganese, aluminium, strontium, nickel and selenium diseases like severe alkalosis [34]. The chlorine content of were detected in traces in beads, indicating bead are richer Rudraksh fruit pulp and bead was found to be 200 & 30 in trace elements, which supports its ethno-medicinal mg/100 g respectively. Pulp contains more level of importance and nutritional value. The other elements, in chlorine as compared to bead and may be a potential descending order, included magnesium, sodium, candidate for chlorine supplement (Table 1). manganese, zinc, iron and copper (Table 2).

Sulphur: Sulphur is the found in sulphur containing Iron: Iron is the most abundant essential trace element in amino acids like methionine, cysteine, homocysteine of humans. It is required for hemoglobin biosynthesis, proteins. It help in amino acids bonding and for normal functioning of CNS and oxidation of biosynthesis of protein. It is mainly found in skin, joints, carbohydrates, protein and fats. Iron is required for the hair and nails. Its intake is helpful in arthritis for repair of synthesis of oxygen transport proteins i.e. hemoglobin the cartilage and in liver disorders. Insulin hormone is a and myoglobin, heme enzymes and other iron-containing protein rich in sulfur-containing amino acids. Glutathione enzymes (Flavin-iron enzymes, transferring and ferritin). sulfur-containing antioxidant, useful in detoxification Two third of total body iron is found in the hemoglobin during metabolism. Bile acid contains taurine, a sulphur present in RBC, 25% iron store in various body tissues containing amino acid useful in digestion. Daily intake of and the remaining 15% in myoglobin [37, 38]. Iron content sulphur is about 800 to 900 mg per day [35]. The sulphur of Rudraksh pulp and bead was found to be 0.082 & 80 content of Rudraksh fruit pulp and bead was found to be mg/100 g respectively. This indicates that Rudraksh bead 110 & 40 mg/100 g respectively indicating the nutritional is a rich source of Iron. importance of pulp as a good source of sulphur (Table I). Zinc: Zinc is the second most abundant transition metal Magnesium: Magnesium functions as a cofactor of many found inside the body after iron and appears in almost all enzymes involved in energy metabolism; protein, RNA & enzyme classes. It is an essential trace element which DNA synthesis. It maintains the electrical potential of plays a major role in various cell processes including nervous tissues and cell membranes. Low serum and normal growth, brain development, bone formation and dietary magnesium may be related to the etiologies of wound healing [38]. In plasma, zinc is transported by cardiovascular disease, hypertension, diabetes and albumin (60%) and transferrin (10%). Transferrin also atherosclerosis [33]. The magnesium content of Rudraksh transports iron, excessive iron can reduce zinc absorption pulp and bead was found to be 80 & 60 mg/100 g and vice versa. Zinc is stored in various body organs i.e. respectively indicating the nutritional importance of fruit prostate, eye, brain, muscle, bones, kidney and liver etc. as a good source of magnesium (Table 1). It plays a vital role in protein & carbohydrate metabolism, help in mobilizing vitamin A from its storage site in the Sodium: Sodium regulates blood volume, blood pressure, liver and facilitates the synthesis of DNA & RNA osmotic balance and maintains constant pH. It is essential necessary for cell growth [39]. It may be used in the for the functioning of neurons and osmoregulation treatment of viral infections, including AIDS, prostate between cells and the extracellular fluid. The presence of gland enlargement, rheumatoid arthritis, wound healing, sodium allows to retain water and helpful to maintain fluid acne, eczema and stress [37]. The trace element analysis volume. It also activates angiotensin II, a protein that revealed the presence of zinc in Rudraksh fruit pulp and constricts arteries to boost blood pressure. It supports bead 120 & 180 ppm respectively indicating the nerve functioning by playing a vital role on action importance of fruit as zinc supplement (Table II). potential. It allows small intestine to absorb amino acids, glucose and water from the foods. It also affects kidney Copper: Copper is the third most abundant trace element function, helping to retain nutrients in the body [33, 34, i.e. 75-100 mg present in human body. It is mainly 36]. Elemental analysis revealed the presence of sodium 10 distributed between the erythrocytes and blood plasma. mg/100 g in Rudraksh bead only (Table 1). It is present in most of the body tissues and stored mainly

624 Middle-East J. Sci. Res., 26 (6): 621-629, 2018 in liver along with the brain, heart, kidney and muscles Selenium: The trace element selenium is an important [40]. Out of total copper reserve, 90% found in component of antioxidant enzymes, glutathione peroxides ceruloplasmin which is responsible for its transport to and thioredoxin reductase. It is an essential trace element, various body cells. Copper-zinc metalloenzyme superoxide which is component of selenoproteins found in enzymes, dismutase contains 60% of copper in RBC and the glutathione peroxidase and other peroxidases. It helps in remaining as loosely bound to other proteins & amino biosynthesis of thyroid hormone and active acids [34, 36]. The trace element copper was found to be immunomodulator, much more potent anti-oxidant than 140 & 120 ppm in the Rudraksh fruit pulp and bead vitamins E, C & A and beta-carotene. It as an antioxidant respectively indicating the nutritional importance of fruit which protect vascular endothelium, low-density (Table 2). lipoproteins, protection of DNA, chromosomes. Selenium helps in protection from atherosclerosis, coronary Aluminum: Aluminium is potent aggregators of ischemic disease and cancer [46]. The trace element bio-molecules and its biological function includes analysis showed the presence of 60 ppm of selenium in significant role in biomolecular compaction. In some Rudraksh bead (Table 2). pathological conditions also molecules are ectopically aggregated by aluminium. Aluminium biocompaction may Molybdenum: Molybdenum is a component of be useful in the condensation of A+T-rich chromatin molybdoprotein, which is essential for formation of active domain and in silencing the expression of some specific sites of various metallo-enzymes. These enzymes help in genes [41]. The trace element analysis revealed that purine catabolism, effect protein synthesis and growth of Rudraksh bead contains 20 mg /100g aluminium (Table 2). the body tissues. Molybdenum containing enzymes are xanthine dehydrogenase/oxidase, aldehyde oxidase and Manganese: Manganese acts as a cofactor in many sulphite oxidase [47]. The trace element analysis of enzymes that are essential for energy production and are Rudraksh fruit pulp and bead shows presence of 140 and involved in the metabolism of glucose, glycogen storage 160 ppm of molybdenum respectively (Table 2). in the liver, protein digestion, synthesis of cholesterol & fatty acids. It plays a vital role in the synthesis of genetic Rubidium: Rubidium has no reported biological effect but molecule DNA and RNA. It is essential for the growth and has a slight stimulatory effect on metabolism, similar to maintenance of the nervous system, the development and potassium. These two elements occur in minerals and maintenance of bones & joints, the function of female sex soils, though potassium abundance is much more than hormones and thyroid hormones. rubidium. During stress or deficiency of potassium some An antioxidant enzyme superoxide dismutase which plants, such as sugar beet, absorb rubidium from the soil. provides protection against free radicals damage also Rubidium enters the food chain and contributes to daily contains manganese [42]. In addition, this enzyme intake 1-5 mg [48]. The trace element analysis revealed the protects brain cells from damage in stroke & Alzheimer presence of 230 & 260 ppm of rubidium in Rudraksh fruit disease [43]. It also protects the liver from damage in pulp and bead respectively (Table 2). alcoholic patients [44]. The trace element analysis revealed the presence of manganese in Rudraksh bead Palladium: Palladium is commonly used in dental casting 11mg /100 g (Table 2). alloys, due to gold cost. However, there have been controversies, over possible adverse biological effects of Chromium: Trivalent Chromium is an essential trace palladium in dental products. Palladium allergy may occur element, playing an important role in glucose metabolism in individuals sensitive to nickel [49]. Rudraksh fruit pulp by serving as a cofactor for insulin action. Hexavalent and bead were found to contain 150 & 260 ppm of chromium is a toxic industrial pollutant and has been palladium respectively (Table 2). classified as carcinogenic. The total body content of chromium is relatively low and is about 0.006g in an Silicon: Silicon is a bioactive element found in high average healthy human adult. It is excreted mainly in the abundance in soil and reaches in biological system urine & feces and in small amount in hairs, sweat and bile through plants. It plays an important role in growth & [45]. The trace element analysis revealed the presence of development of plants and also protects plant tissues chromium in Rudraksh fruit pulp is very low i.e. 9 ppm from various biotic & abiotic stresses. Silicon strengthens (Table 2). bones and improves immune response, also found in

625 Middle-East J. Sci. Res., 26 (6): 621-629, 2018 connective and nerve tissues [50]. This element is found needed only in trace amounts, enables body to use in abundance 30 & 70 mg/100 g in Rudraksh fruit pulp and vitamin A to keep eyesight healthy. Vitamin D helps body bead respectively (Table 2). more readily to absorb calcium [57]. A deficiency in major or trace mineral can lead to Arsenic: Arsenic may cause heritable changes in gene poor health and serious illness that affects body systems. expression, including DNA methylation, histone A calcium deficiency can lead to osteoporosis or weak, modification and RNA interference without changing brittle bones as well as impaired heart function. A DNA sequence. Toxic levels of arsenic cause significant deficiency of copper, a mineral needed in trace amounts, DNA hyper methylation increasing risk of carcinogenesis can lead to slowed growth in babies, low immune function [51]. The trace element analysis revealed the presence of and changes in the bone marrow. 40 ppm arsenic in Rudraksh fruit pulp (Table 2). The essential elements magnesium, potassium, calcium, phosphorous, sulphur and chloride were Bromine: Bromine is a cofactor for enzymes involved in determined in bead and pulp powder of Elaeocarpus post-translational modification of proteins and it is ganitrus indicates its nutritional importance. The daily essential for growth and development of tissues. It is an requirement of macro-elements for an adult person is essential trace element for all organisms and its deficiency above 100?mg/day [58]. The pulp was found to be very may be cause basement membrane alterations in rich in potassium (1470 mg / 100 g), followed by calcium nutritional deficiency and smoking patients [52]. The trace (240 mg / 100 g) in bead. Trace elements iron, copper, zinc, element analysis revealed the presence of 60 ppm bromine molybdenum, palladium, rubidium and silicon were in Rudraksh fruit pulp (Table 2). detected in both bead and pulp. Chromium, bromine and arsenic were found in traces in fruit pulp. However Strontium: The trace element strontium is reported as manganese, aluminium, strontium, nickel and selenium potential therapeutic agent for osteoporosis. Recent were detected in traces in beads. These trace elements are reports shows that low doses of strontium have beneficial essential prosthetic group of many metaloenzymes effect on bone formation in osteoporotic patients [53]. involved in body metabolism and various cellular Strontium promoting action on calcium uptake into bone processes. Hence the consumption of whole fruit may at moderate strontium supplementation [54]. The trace fulfil the body’s requirement of essential & trace elements element analysis shows 110 ppm of strontium in Rudraksh for metabolism, growth and development. The availability bead (Table 2). of the plant in Indian subcontinent may be used as a source of nutrients especially in developing countries in Nickel: The most of studies on the role of nickel are done the region. on animals, its relevance in humans yet to be verified. Nickel is generally distributed in the body slightly in ACKNOWLEDGEMENT larger amount in the nucleic acid, especially in ribonucleic acid (RNA). It is considered to affect the structure or We are thankful to Mahamandaleshwar Swami function of the proteins associated with nucleic acids. Maartand Puri, Chairman, Centre for Spirituality Research Nickel is also found in enzymes involved in glucose and Studies, Shobhit University, Gangoh, Saharanpur, UP, metabolism. It also helps in production of milk in the India for his valuable guidance and support during the mammary glands by production of prolactin hormone. studies. Nickel also affects overall growth, healthy skin and bone structure. It helps in absorption of iron from food and Conflict of Interest plays an important role in RBC production [55, 56]. None. The trace element analysis revealed the presence of 70 ppm of nickel in Rudraksh bead (Table 2). REFERENCES

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