Effect of Varying Environmental Conditions on Mineral Status of Stem Bark of Anogeissus Latifolia

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Udaysing Hari Patil et al. / Journal of Pharmacy Research 2012,5(2),1140-1143 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Effect of varying environmental conditions on mineral status of stem bark of Anogeissus latifolia

Udaysing Hari Patil*1 and Dattatraya. K. Gaikwad2 *1Department of Botany, Bhogawati Mahavidyalaya, Kurukali. Tal-Karveer, Dist- Kolhapur [MS] India-416001. 2Department of Botany, Shivaji University Kolhapur, Maharashtra, India-416004 Received on:10-11-2011; Revised on: 15-12-2011; Accepted on:12-01-2012

ABSTRACT Anogeissus latifolia is used in various cases of Heart troubles. Present study deals with the investigation of different bark samples (Apical bark, middle bark and mature inner bark) of Anogeissus latifolia for inorganic nutrients. All the bark samples contain appreciable amount of key elements Nitrogen (0.69-1.77%), Phosphorus (0.031-0.103%), Calcium (0.94-1.46%), and Magnesium (0.134-0.223%), minor elements Copper (1.02-2.02mg/100g), Zinc (1.22-1.9mg/100g), Manganese (0.92-1.36mg/100g) and Iron (10-10mg/100g) and heavy elements Chromium (0.46-1.76mg/100g), Nickel (0.18-1.38mg/ 100g) and Cadmium ((0.06-0.24mg/100g). Lead was found absent in all the bark samples analyzed.

Key words: Anogeissus latifolia, Inorganic a status, macro-minerals, traces minerals, heavy metals.

INTRODUCTION Anogeissus latifolia, belonging to the family Combretaceae is medium sized colour was measured at 520nm on a double beam UV- spectrophotometer deciduous tree [1]. Grey-white bark is smooth and is effective in anemic (Shimadtzu UV-190) after 15 minutes. Total nitrogen was calculated by conditions, urinary discharges, and piles [2, 3]. Stem bark is useful in diar- using standard curve obtained by using different concentrations of standard rhea, dysuria, colic, liver complaints, snakebite and skin diseases [4]. The ammonium sulfate solution and employing the similar procedure as de- bark of this tree is useful in the treatment of fever [5]. Decoction of bark is scribed for the analysis of samples. remedy against cough [6]. Many of the inorganic constituents contribute equally as the plant secondary metabolite (phytochemicals) in the defense Preparation of acid digests mechanism against harmful diseases and at the same time many put in Acid digestion method developed by Toth et al.[8] was followed for the certain nasty diseases. This fact enabled to undertake the standardization analysis of inorganic constituents. 0.5g oven dried bark powder was trans- of this crud drug for macro, micro and heavy elements. ferred to 100ml capacity beaker and 20ml concentrated HNO3 was added to it. The beaker was kept covered with watch glass till the primary reactions MATERIALS AND METHODS completed. The beaker was heated gently on hot plate to dissolve solid particles of bark powder. After cooling to room temperature, 60%, 10ml Total nitrogen content perchloric acid was added to it and mixed thoroughly. The beaker was again Total nitrogen from the bark was estimated according to the method de- [7] heated strongly until a clear 2-3ml colorless solution was obtained. The scribed by Hawk et al. . Plant material, 0.5g oven dried powdered bark beaker was cooled and contents in the beaker were diluted to 100ml with was taken in Kjeldahl’s flask. To this a pinch of microsalt (200g K SO + 5g 2 4 distilled water and kept overnight. On the next day, acid digest was filtered CuSO dehydrated) and 5ml H SO (1:1) was added. To avoid bumping few 4 2 4 through ashless filter paper (Whatman No.44) and filtrate was used for glass beads were added to the flasks and the material was digested on low estimation of different inorganic constituents. flame. Faint yellow coloured solution obtained after complete digestion cooled to room temperature and diluted to 100ml with distilled water. The Phosphorus content solution was filtered through Whatman No. 1 filter paper and used for The phosphorus content was estimated according to the method developed estimation of total nitrogen. by Sekine et al.[9]. Phosphorus reacts with molybdate-vanadate reagent (MV reagent) to give yellow colour complex. The intensity of yellow 2ml plant extract was taken in a set of Nessler’s tubes. To each this tube, coloured complex was estimated by using spectrophotometer and by com- one drop of 8% KHSO was added and volume was adjusted to 35ml with 4 paring with the colour intensity of the known standards, phosphorus con- distilled water. 15ml of freshly prepared Nessler’s reagent (Reagent A: 7g tent was estimated. KI + 10g HgI2 in 40ml distilled water, Reagent B: 10 NaOH in 50ml water. ‘A’ and ‘B’ were mixed in proportion of 4:5 only at the time of estimation) 2ml acid digest was taken in the test tube and equal amount of 2N HNO3 was added to each test tube. The reaction between sample and the reagent was added followed by 1ml freshly prepared molybdate-vanadate reagent. gave orange brown coloured product of NH Hg I . The intensity of this 4 2 3 Final volume in test tube was adjusted to 10ml with distilled water. The ingredients were mixed well and allowed to react for 20 minutes. After 20 minutes, colour intensity was measured at 420nm using a blank containing no phosphorus. Total phosphorus was calculated with help of standard *Corresponding author. curve obtained by using different concentrations of standard phosphorus Dr. Udaysing Hari Patil. solutions. Other steps are essentially similar as described above. Phospho- Assistant Professor, rus in the plant material was expressed in g.100g-1 on dry weight basis. Department of Botany, Bhogawati Mahavidyalaya, The levels of Calcium, Magnesium, Iron, Manganese, Copper, Zinc, Chro- Kurukali. Tal-Karveer mium, Cadmium, Nickel and Lead were estimated using atomic absorption Dist- Kolhapur -416001 spectrophotometer (AAS-Model Perkins Elmer 3030). In case needed, ap- Maharastra, India propriate dilutions of plant extract were made with distilled water.

Journal of Pharmacy Research Vol.5 Issue 2.February 2012 1140-1143 Udaysing Hari Patil et al. / Journal of Pharmacy Research 2012,5(2),1140-1143 RESULTS AND DISCUSSION material, DNA and RNA of all cells. In the recent analysis apical stem bark Results of the present study are demonstrated in the Fig. 1, Fig. 2 and Fig. (0.103%) reported to contain higher amount during summer whereas, low- 3. Nitrogen is integral part of the nucleic acids DNA, RNA, proteins, amino est amount was quantified in mature inner bark (0.031%) during winter. In acids, some of the plant growth regulators such as IAA, cytokinins and case of middle bark, a total Phosphorus content was moderate (0.069- vitamins. Amount of total nitrogen content determined was higher in the 0.084%). These vales are significantly lower than phosphorus levels esti- apical stem bark (1.77%) and lowest in mature inner bark (0.69%) during mated in Lettuce (0.98%) and Spinach (0.95%) by Davidson and Le Clerc summer and during winter respectively. Total nitrogen share in the middle [11]. Calcium occurs as free Ca+2 as divalent cation in plants. It is also present bark was moderate and ranged from 0.89% to 1.41%. Total nitrogen content in the form of calcium oxalate, carbonates and phosphates (Mengel and determined in the bark of black locust (3.43% to 3.45%) [10] exceeds the Kikby, 1968)[12]. Calcium Deficiency causes osteoporosis [13]. In the present nitrogen content determined in present inspection. Phosphorus is both investigation higher Calcium concentration was determined in the mature obligatory for plant and human, is often referred as “energizer”, since it inner bark (1.46%) during winter and the lowest and reduced amount was helps to assemble and shuffle energy. It is also a constituent of genetic recorded in apical stem bark (0.94%) during winter, while, calcium content in middle bark was in between apical and mature bark. Calcium content in the middle bark was found to be fluctuated between 1.17 -1.40%. These values are lower than the Cinnamomum ovalifolium (9.99%) [14]. Magnesium is strongly electropositive, mobile element in the plant and average requirement for optimal plant growth varies 0.5%-1.0%. It functions as a cofactor for enzymes involved in protein synthesis, RNA and DNA synthesis, and maintenance of the electrical potential of nervous tissues and cell membranes [15]. Quite higher amount of Mg was recorded in apical bark (0.223%) during summer when compared with lowest level in mature inner bark (0.134%) during summer followed by moderate peak in middle bark (0.14-0.167%). The bark of Cinnamomum ovalifolium (1.915%) [14] contains much higher Mg than the bark of Anogeissus latifolia in the present investigation. Fig.1. Quantitative estimation of macro elements (N, P, Ca and Mg) from Anogeissus latifolia bark Copper is essential for proper functioning of copper-dependent enzymes [16]. Copper deficiency increases risk of cardiovascular diseases [17]. Apical bark (2.02mg/100g) of Anogeissus latifolia was reported to accumulate maximum copper during summer followed by least concentrated in mature inner bark (1.02mg/100g) during winter and fair in middle bark (1.24-1.34mg/100g). Copper reported in present study is in the array of Cinnamomum ovalifolium, (1.42mg/100g) [14]. Zinc is ubiquitous in plants, mi- croorganisms, and animals. The average adult ingests 10 to 15mg of zinc daily. Severe zinc deficiency in humans results in growth retardation, delayed sexual and bone maturation, skin lesions, diarrhoea, alope- cia, impaired appetite and the appearance of behav- ioral changes [18]. No large differentiations were ob- served over the changing climatic conditions in each bark sample. Zinc content in the apical bark (1.9mg/ Fig.2. Quantitative estimation of micro elements (Cu, Zn, Mn and Fe) from Anogeissus latifolia bark. 100g) was similar and highest during both seasons, in the middle bark 1.3-1.8mg/100g and lowest in mature inner bark (1.22-1.24mg/100g). Zinc estimated in Alternanthera pungens (3.786mg/100g)[19] is higher than diagnosed in the recent examination. A minimum intake of 2.5 to 7 milligrams of manganese per day meets human needs [20]. Manganese is involved in the formation of thyroxin, is required for normal thyroid function [21]. Manganese deficiency in human cause hypocholesterolemia, decrease in triglyc- erides and phospholipids, weight loss, transient der- matitis, and intermittent nausea [22]. A high concentration of Mn was reported in the apical stem bark (1.6mg/100g) during summer than middle and mature inner bark. Manganese concentration in middle bark (0.96-0.98mg/100g) and mature inner bark (0.92- 0.94mg/100g) was fairly constant during both sea- Fig.3. Quantitative estimation of heavy elements (Cr, Ni, Cd and Pb) from Anogeissus latifolia bark. Journal of Pharmacy Research Vol.5 Issue 2.February 2012 1140-1143 Udaysing Hari Patil et al. / Journal of Pharmacy Research 2012,5(2),1140-1143 sons. Manganese reported in the leaves of Lettuce (0.288mg/100g) and 2008; Pp166. Leek (0.173mg/100g)[23] is lower than the Mn content reported in the present 7. Hawk PB, Oser BL, Summerson WH. Practical Physiological chem- study. Iron, is one of the most important micronutrient in plant and ani- istry. Publ. The Blackiston Company, USA. 1948; 120-121. mals. Early iron deficiency leads to irreparable damage to brain cells, phago- 8. Toth SJ, Prince AL, Wallace A, Mikkelsen DS. Rapid quantitative cytosis and bacteria killing functions are impaired in iron deficiency [24]. determination of 8 mineral elements in plant tissues by systematic Highest iron content was determined in the apical bark (20.84mg/100g) procedure involving use of a flame photometer. Soil Sci,.1948; 66: during winter and minimum value was recorded in mature inner bark (10mg/ 456-466. 100g) during summer. The middle bark iron composition was 12-17.6mg/ 9. Sekine T, Sasakawa T, Morita Kimara, T, Karatomi K. Laboratory 100g). These values are much lower than Brassica campestris (188.97mg/ manual for physiological studies of Rice. Yoshida S, Forno D, Cook 100g) [19]. JH, Gomez KA. (Eds). International Rice Research Institute, Ma- nila 1965; 45-46. Chromium (Cr) is not naturally present in the earth crust. Trivalent chro- 10. Jones DB, Phillips S. Protein content of the bark of black locust, mium (Cr3+) is most found in living organisms. Cr plays important role in Robinia pseudacacia. J. Am. Chem. Soc., 1937; 59 (3):595–596. 7 lipid metabolism, reducing the risk of atherogenesis. Chromium supple- 11. Davidson J, Eclebc JAL. The variation in the mineral content of mentation increases HDL-cholesterol [25] and decreases total cholesterol, vegetables. Journal of nutrition., 1994; 23:55-66. 8 LDL-cholesterol and triacylglycerols [26] in humans. Chromium toxicity 12. Burstrom HG. Calcium and plant growth. Biological Reviews, 1968; causes oxidative DNA impairment [27] and genotoxicity [28]. The Chromium 43(3):287–316.9 concentration among the three bark samples was higher in the mature inner 13. Nordin BEC. Osteomalacia, osteoporosis and calcium deficiency. bark (1.76mg/100g) during summer and drops down to lowest amount in Journal of Clinical Orthopaedics and Related Research, 1960; apical bark (0.46mg/100g) during summer. Cr concentration fluctuated from 17:235–258.10 0.92-1.58mg/100g for middle bark. Chromium content in spinach (0.660mg/ 14. Chandrajith R, Nadeesha Koralegedara KB, Ranawana HJ, Tobschall 100g)[29] and clover shoot (0.265mg/100g)[29] are lower than Cr amount re- CB. Major and trace elements in plants and soils in Horton Plains ported in present work. In biological systems, Ni+2 form of Nickel is the National Park, Sri Lanka: An approach to explain forest die back. most common [30]. Nickel is heavy metal, short term expose also causes Environ Geol., 2009; 57:17–28. 11 nausea headache, vomiting [31]. While evaluating concentration of nickel in 15. Classen HG. Magnesium and potassium deprivation and supplemen- the bark of Anogeissus latifolia, nickel concentration was increased from its tation in animals and man: aspects in view of intestinal absorption. lowest level 0.18mg/100g in apical bark during summer to maximum in Magnesium, 1984; 3:257–264. 12 mature inner bark ((1.38mg/100g) during winter. Nickel was determined in 16. Solomons NW. Biochemical, metabolic, and clinical role of copper moderate concentration in middle bark (0.34-1mg/100g). These values lower in human nutrition. J Am Coll Nutr 1985; 4(1):83-105. 13 than reported in Cannabis sativa (1.58mg/kg) and Hordeum vulgare 17. Klevay LM, Inman L, Johnson LK, Lawler M, Mahalco JRD, Milne (1.41mg/kg)[19]. Cadmium is regularly found in ores together with zinc, B. Increased cholesterol in plasma in a young man during experi- copper and lead. Exposure to high Cd toxicity cause bone damage [32] and mental copper depletion. Metabolism, 1984; 33:1112–1118. 14 develop acute respiratory distress syndromes (ARDS) [33]. In the present 18. Hambridge KM, Casey CE, Krebs NF. Zinc Trace elements in hu- test, mature inner bark (0.24mg/100g) showed upper limit of cadmium man and animal nutrition, 5th ed. Academic Press, 1987; 2:1–137. during summer and smallest amount in apical bark (0.06mg/100g) during 19. Jabeen N, Ahmed M. Possible allelopathic effect of three different winter, while, moderate in middle bark (0.08-0.12mg/100g). These values weeds on germination and growth of maize (Zea maize) cultivars. exceed the values reported for spinach shoot (0.023mg/100g) [29]. According Pak. J. Bot., 2009; 41(4): 1677-1683. 16 to WHO, exceeding 0.3mg/Kg of lead which is threshold limit for safety, 20. Hegsted B M. The presence of tightly bound manganese. Biol. results in lead poisoning, blindness, deafness, hypertension, impairment of Chem., 1976; 241:196-207. kidney function and neurological disorder[34]. All the bark samples of 21. Pfeiffer CC. Mental and Elemental Nutrients. Keats Pub. Conn. In: Anogeissus latifolia under investigation showed absence of lead (Pb). Phosphate and magnesium metabolism. Edinburgh, Churchill Livingstone, 1976; 1–35. 18 CONCLUSION 22. Doisy CA. 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Journal of Pharmacy Research Vol.5 Issue 2.February 2012 1140-1143