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Comparative Study of Metal Uptake by Medicinal Plants Growing In Natural and Polluted Area of Eastern Ghats
Sr. Prema Kumari1*, K Joanny1, Louis Jesudas2, Raj Kumar, S. D. 3 1 St. Anns College for women, Malkapuram, Visakhapatnam-530011, AP; 2 Department of Plant Biology &Plant Biotechnology, St. Xaviers College, Palayamkottai, Nellai627002, TN; 3 St. Andrews PG College, Department of Botany, Gorakhpur-273001;
Metal pollution has become a major growing concern in the industrial area which is surrounded by many industries like Coromandel fertilizers, Hindustan Petroleum Corporation Limited, Hindustan Shipyard, Steel Plant etc. along the coast. Increasing industrial activity accumulated various heavy metals in varied degrees in the soil. The uptake of these metal pollutants is analyzed in the medicinal plants exhibiting luxuriant growth in the polluted sources as well as in natural sources. This survey focuses on heavy metals, that are essential (Cu, Co, Fe, Mo, Ni, Zn), beneficial (Na, Co etc.) and toxic (Cd, Pb, Hg). Keywords: Heavy metals, uptake, Industries, medicinal plants ICP MS.

INTRODUCTION
Of the 109 known elements, relatively a small fraction of elements are known to be biologically important. Natural abundance limits the availability of the elements for such use. Life evolved utilizing those elements that were abundant and available to it. Those elements that were rare were not utilized by the living system as they are not available; neither the system evolved the mechanism to cope with them. Among an array of heavy metals, Cu, Co, Fe, Mo, Ni and Zn are essential micronutrient mineral elements, whereas Cd, Pb, Hg, As etc. Have no known physiological function in plants and are potential toxins to living organisms (21 and 22). Large residential areas, especially near industrial belt are contaminated by heavy metals that mainly originate due to industrial activity in Malkapuram cluster. However, elevated levels of both essential and non-essential heavy metals pose serious threat for human health and agriculture. The excessive uptake of these metals from the soil can create dual problem: Firstly the harvested crops so contaminated serve as a source of heavy metals in our food supply, and secondly yields are reduced due to adverse effect on plant growth (Bala and Setia, 1990; Hall, 2000). These non-degradable heavy metals are important environmental insidious pollutants and also a cause of potential ecological risk (4). Heavy metals are well known to be toxic to most organisms when present in excessive concentrations. First observations of the effects of heavy metals on soil microbial processes date back to the beginning of this century2. But only when the large adverse effects of emissions of heavy metals from smelters on surrounding ecosystems was observed in the 1960-70s it was realized how severely soil microorganisms and soil microbial processes can become disrupted by elevated metal concentrations, sometimes resulting in severe ecosystem disturbance (1). In Visakhapatnam area itself there are 14 major hazardous industries are present among which 7 are present in Malkapuram cluster that has been selected

for our study. The industries are namely HPCL (R), HPCL (T), IOCL (T), LPG bolting plant, CFL, APCL and BPCL.

However, toxic metal pollution of the biosphere has intensified rapidly since the onset of the industrial revolution, posing major environmental and health threats (3). Many elements in our analysis are released by the industries that are mentioned above (20). There are 35 metals that concern us because of occupational or residential exposure; of these 26 elements are included in our study. Interestingly, small amounts of these elements are common in our environment and diet, and are actually necessary for good health, but large amounts of any of them may cause acute or chronic toxicity (poisoning). For some heavy metals, toxic levels can be just above the background concentrations naturally found in nature. Therefore, it is important for us to inform ourselves about the heavy metals and to take protective measures against excessive exposure. Our study concentrates on 26 elements which includes both heavy metals, essential and non-essential elements (11). Heavy metals become toxic when they are not metabolized by the body and accumulate in the soft tissues. Industrial exposure accounts for a common route of exposure for adults. However in plants the toxic range of elements may be altogether different as plants respond to abiotic stress through secondary metabolite elicitation. In general to determine the hazardous association with heavy metal contamination of the environment, the most sensitive (crucial) step is to determine the most sensitive environment organism. In contrast we have selected the medicinal plants thriving well in most polluted industrial area to study the effects of metal contamination. The earths crust is the ultimate source of all metallic elements found in the environment. Metals are neither created nor destroyed, but are redistributed by geobiological cycle. Due to industrial

Corresponding Author: Sr. Prema Kumari, St. Anns College, Malkapuram, Visakhapatnam; Received 18-11-2011; Revised 21-02-2012; Accepted 20-04-2012
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Sr. Prema Kumari, et al.: Comparative Study of Metal Uptake by Medicinal Plants Growing In Natural and Polluted Area of Eastern Ghats

activities of ore phase of a metal shortens and forms new metallic compounds are released into the atmosphere. Heavy metal accumulation in the crop plants may have adverse effect on food safety, phytotoxicity and environmental health of soil organism. This article details the uptake of heavy metals by various medicinal plants grown in polluted areas and natural resources. The study includes 10 plants out of which few plants are specific to respiratory diseases and others are not very specific for respiratory problems. These plants that are subjected to metal stress may trigger the secondary metabolites of pharmacological importance which in turn may be beneficial to the mankind.

Dwell Time Integration Time Replicates Internal Standard 103 Rh

50 ms 2500 ms 3

Materials: A hot plate with digital temperature controller with a maximum temp of 250oC was used for digestion. Teflon beakers thoroughly cleaned, soaked in 1:1HNO3 for 6 hours and thoroughly rinsed with milliQ water were used for digestion. Thoroughly acid cleaned 100ml and 250ml standard flasks were used for volume make up. Whattman filter paper no.42 was used for filtration purposes. Determination of Trace Metals in Plant: Open acid digestion method was followed for the determination of metal concentration, wherein representative samples of dried plant tissues (approximately (~0.5g) were taken in Teflon beakers and 30ml conc. HNO3 were added in each. They were heated on hot plate (~1000C) for two hours keeping the lids. At boiling stage, about 4-5ml H2O2 was added drop-wise and heated further and the volume was reduced to about 10ml. During this entire process all organic material gets oxidized and the inorganic contents are extracted into the solution. To this 5 ml of 1g/g Rh solution was added to act as an internal standard and the solution was transferred to 250ml of volumetric flask and diluted to 250 ml with Milli-Q water. The solution was analyzed by ICP-MS for trace elements. Determination of Trace Metals in Soil: About 0.05g soil samples were weighed and taken in a clean PTFE Teflon beaker. Each sample was moistened with a few drops of water. Then 10mL of the acid mixture containing a 7:3:1 ratio of HF, HNO3, and HClO4 was added to each beaker and the sample swirled until completely moistened. The beakers were covered with lids and the samples left standing overnight after adding 5mL of 103Rh (1g/ml) as an internal standard. The next day, the beakers were heated on a hot plate at 220oC for about 1 hour, after which the lids were removed, the contents evaporated to near dryness. The evaporation process was repeated after adding 5mL of the above acid mixture in each case. Finally, the residue was dissolved by gently heating in 20mL of 1:1 HNO3. Clear solutions were obtained for all samples. After cooling to room temperature, the volume was made up to 250ml, and these final solutions were stored in polyethylene bottles. The concentrations of different metals in these solutions were analyzed by ICP-MS. International geochemical standards SO-1 and SO-2 was used for calibration as well as to check the accuracy and precision.

MATERIALS AND METHODS

Plants Used: Abutilon indicum (L.), Achyranthes aspera var. perphyristachya Hook. F., Adhatoda vasica Nees, Alternanthera sessilis DC, Azadirachta indica A. Juss, Recinus communis Linn, Eucalyptus globulus (LABILLE), Hyptis pectinata (L.), Ocimum sanctum Linn and Tinospora cordifolia Chemicals Used: water HF, HNO3, HClO4, H2O2 and MQ

Instrument: ICP-MS (PerkinElmer Sciex Instrument, model ELAN DRC II, Toronto, Canada) Experimental: Reagents and samples: All chemicals were of analytical reagent grade and were supplied by Merck (Darmstad, Germany). Concentrated HNO3 (65%) was used for acid digestions. This high purity Nitric acid was used to minimize the procedural blank values. Ultra pure water was prepared by passing doubly deionized water from a milliQ system (Millipore, USA) and was used throughout the analysis. Two certified reference materials (NIST 164Oa and NIST 1643e) were used for calibrating the system as well for checking the accuracy of the data. Instrumentation: A Perkin Elmer SCIEX model Elan DRC II ICP-MS (Ontario, Canada) at CSIR-National Geophysical Research Institute, Hyderabad, was used throughout for trace elements analysis of plant and soil samples. The sample introduction system consisted of a standard Meinhard nebulizer with a cyclonic spray chamber. Instrumental and data acquisition parameters have been listed in table.1. ICP-MS Instrumental and Data Acquisition Parameters Instrumental Parameters
RF Power Argon Gas Flow Nebulizer Auxiliary L/min Plasma Lens Voltage Sample Uptake Rate Data Acquisition Parameters 1100 W 0.84 L/min 1.20 15 L/min 7.75 V 0.8mL/min Quantitative Mode 1

RESULTS AND DISCUSSION

Eastern Ghats is known for rich vegetation in terms of medicinal plants. We have selected few plants such as Ocimum, Eucalyptus, Tinospora and Adathoda exhibiting luxuriant growth and are known to have curative action on respiratory ailments along with other general medicinal plants. To study the effect of metal pollution on curative action in the industrial area of Eastern Ghats, on above mentioned medicinal plants, it is necessary to distinguish between uptake arising from natural metal sources and from pollutant sources. Metals from both natural and pollutant sources have the potential for being assimilated by the plant through

Measuring Mode Peak Hopping Point per Peak Number of Sweeps

50

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Sr. Prema Kumari, et al.: Comparative Study of Metal Uptake by Medicinal Plants Growing In Natural and Polluted Area of Eastern Ghats

foliar or root absorption processes. The presence of heavy metals in medicinal plants at various degrees depends on their oxidations as well as their concentration (25) the plants growing in polluted industrial areas are experimentally showing Na, Fe, Cd, Pb, Al, Zn, V, Mg, Co, Mn, Ni and Cr in more concentration in various plants. The major factor governing availability of elements to plants from soils is likely to be based on the solubility and the thermodynamic activity of the uncomplexed ion (6). For root uptake the soluble species must exist adjacent to the root membrane for some finite period (7). It is generally considered that the uptake of nutrients (essential elements) by plants is metabolically regulated, at the same time the mechanism controlling the absorption of non nutrients species beneficial elements is controversial. Table I: Analysis of Metals in Soil Samples (concentration in ppm)
Soil Elements As Ba Co Cr Cu Mo Ni Pb Rb Sr V Y Zn Zr SiO2 A1203 Fe203 MnO MgO CaO Na2O K2O TiO2 P2O5 Natural Source 5.35 519.4 6.05 57.8 ND 5.2 ND 31.8 176.4 86.25 43.4 49.7 71.65 216.25 84.4 8.2 1.7 0.06 0.59 ND 1.1 11.9 0.68 0.1 Polluted Source 6.2 857.5 27.3 211 33.6 ND ND 43 44.5 104.2 181.4 25.7 338.8 177.3 53.6 6.38 6.19 0.16 0.8 3.26 1 4.88 1.86 1.79

Twenty four elements have been studied in the soils of natural and polluted soils using ICP-MS (PerkinElmer Sciex Instrument, model ELAN DRC II, USA). Out of them nine essential trace elements (Fe, Co, Mn, Cu, Mo, V, Cr, Mn, Ni), Mo is the heaviest (Z=42) and is relatively less abundant in the earths crust. In our analysis of natural and polluted soils, Ni is not detected and Mo is in less concentration in the natural sources and not detected in polluted soils. Cu is not detected in natural soils and more abundant in polluted soils. Zn, Co, Cr and V are more in polluted soils approximately four times. Fe and Mn are in oxide form with no significant increase in polluted soils. Al, Ti and Zr are not essential, yet abundant in earths crust and form extremely insoluble oxides. The presence of Al, Ti and Zr show no significant variation in natural and polluted soils. Analysis of six essential non-metals (C, N, O, P, S, Cl) are not included in the analysis. Si is one of the abundant elements in the earths crust and has been found to be essential probably in one family of plants (30). Si is present at higher range in both the soils and in treated and control plants with not much significant variation. All the representative metals Na, K, Mg and Ca are in oxide form in soils and are found at higher concentration in both the plants grown in natural and polluted sources. Y, Sr, Rb, Pb, Ba, as appear to be non nutrient elements. They are neither included in the above essentiality list nor show significant variation in polluted and natural soils. In polluted soils, Ba is in highest concentration followed by Zn, Cr, V, Zr, Sr, Si, Rb, Pb. Metabolic adaptation as a mechanism of metal tolerance in plants appears to be a rule in plant populations derived from environment containing elevated levels of Lead (28). The uptake of both nutrient elements and non nutrient elements is metabolically regulated as relatively higher concentration ratio values are obtained not only for nutrient species (Mo, Mn, Cu and Zn) but also for non nutrient species such as Pb, Ni, Cd, Ti, As and Sn(29). Analysis of Metals in Plant Samples: A total of 26 elements V, Cr, Mn, Ni, Co, Cu, Zn, As, Se, Rb, Sr, Mo, Ag, Cd, Sb, Ba, Ti, Pb, U, Na, Mg, Al, Si, K, Ca, Fe were estimated in the powdered medicinal plant sample of ten, each of plants grown under natural resources and plants thriving under polluted sources. Out these the role played by 16 elements (Na, Mg, Al, Si, K, Ca, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Mo, as) as macro and micro nutrients to support the growth and development of the plants is established (13). From the study it was revealed that all the metals were accumulated to greater or lesser extent by all plants species studied. The plants showed large number of elements and were rich in Na, Mg, Ca, K, Fe and Al, Si which are earths crust elements, and the same were found to be abundant in earths crust also. The data in table.2 indicate the uptake of elements (macro, trace and heavy elements) by the various medicinal plants grown in natural and pollutant areas and their comparative analysis is drawn.

ND: Not Detected Table No.1 shows the elements that are present in the soil sample of the Malkapuram industrial area and the non-polluted agency area that have been selected for study (Ba, Co, Cr, Cu, Pb, Sr, Zn, Al, Fe, Mn, Mg, Mo, Ti, P are present in higher concentration in the soil of industrial area). This high concentration may be attributed to the release of the elements from the surrounding industries of Malkapuram as the same are not found in control soil samples at higher concentration. In contrast, in control soils Rb, Y, Zr, Si, K are at higher range and Na is equal in both. Of all the 24 elements reported in soils of industrial belt Co. Cr, V, Zn, Fe are almost four folds more than in control soils. We concentrate our study on the uptake of these metals by the medicinal plants of polluted and natural areas. Elements found in soil largely reflect the composition of earths crust all though some modifications occur by further weathering, chemical changes which includes mineral dissolution, while biological activity causes enrichment of Macro elements(15). Factors such as light intensity Oxygen tension and temperature are known to affect the uptake of minerals (23). Operator the energy derived from photosynthesis and Oxygen released can improve conditions for the active & absorption of elements, However interaction between metals is often complex as they are dependent on the metal concentration, PH and growth media (24).

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Sr. Prema Kumari, et al.: Comparative Study of Metal Uptake by Medicinal Plants Growing In Natural and Polluted Area of Eastern Ghats

In all the plants that have been studied for metal uptake six elements namely Na, Mg, Al, Si, K, Ca are found in high concentration in both control and polluted plants ranging from 4ppm to 354ppm and Fe shows in four plants Abutliton, Achyranthes, Alterlanthera and Tinospora respectively ranging between 001 to 5.48ppm. Where as all other 19 elemental falls below the 1ppm in all plants. In the comparative study the plants grown in natural and the polluted areas, out of 26 metal elements detected in all plants and 6 plants ( Abutilon, Alternanthera, Azadirachta, Recinus, Eucalyptus, Ocimum) show more than 10 elements in higher concentration and remaining four plants ( Achyranthes, Adhatoda, Hyptis, Tinospora) with less than 10 elements in higher concentration. Alternanthera, Abutilon and Ricinus are showing accumulation of maximum number of elements 19, 15 and 15 respectively. High element concentration (Plants Alternanthera, Abutilon, Ocimum) and less number is found in Achyranthus, Tinospora 4 and 5 number of elements respectively. The toxic elements like Pb, Cd, Cr, V, As, U and Ba are seen at different folds of increase. Abutilon shows 2 fold increases of toxic elements-V, Co, Cr, Zn and Fe.
Eleme nt Fe Co Zn Cu Mo V Cr Mn Ni 001 1.5368 0.0226 0.5606 3 0.1040 2 0.0053 4 0.0074 5 0.0887 9 0.2980 7 0.0250 6 111 5.4841 3 0.0041 2 1.2336 6 0.1439 1 0.0045 1 0.0173 3 0.1140 3 0.6423 3 0.0512 002 112 003

Pd, Cd are two fold in Achyranthus Pb 100 fold in Adathoda, Cd 3 fold in Alternanthera, Cd, Pb 3 fold high in Azadhirecta, Cd, Pb, U are 2-5 fold in Ocimum, Fe, As 1-5 to 2 fold in Ricinus. Lead accumulation has been found to be in various concentrations in different plants. Pb a heavy toxic metal is molecularly sticky not mobile and therefore not easy to get into plant from the soil (27). Yet it is very interesting from the scientific stand point to find out Pb in 100 fold in Adathoda. Cd has no toxic effect up to 50pp on plants. For concentration higher than 100ppt, plant fresh weight decreased & in plants showing up to 750ppm plant growth has been very difficult. The increase in Cd uptake by plants is associated with cation exchange capacity along with Cd uptake Na uptake has been found to be increasing accordingly. Therefore from the above observation the plants showing more accumulation of heavy metals like Pb, Cd need to be further analyzed in comparison with plants already included in the list of phytoremediation as there show luxuriant growth, and thus these plants may have potential exploitation for soil decontamination.

Table.2a ; Showing metal concentration in 10 natural and polluted plants

113 004 114 005 0.6867 2 0.0094 8 0.5560 1 0.0683 7 0.0253 2 0.0029 9 0.0806 6 4.9884 2 0.0476 7 115 1.0153 6 0.0008 4 0.6660 9 0.0487 9 0.0092 0.0047 2 0.0803 0.6440 6 0.0300 6 1.2655 1.3520 0.9879 1.7741 1.2277 2.9085 5 3 6 2 6 2 0.0022 0.0016 0.0024 0.0019 0.0019 0.0031 4 8 4 2 7 7 0.4970 0.5176 0.6723 0.5714 0.3549 0.8851 2 3 7 5 4 0.0973 0.0821 0.0827 0.0877 0.0790 0.1010 9 8 8 3 1 7 0.0038 0.0028 0.0057 0.0265 0.0040 0.0061 2 5 6 1 6 1 0.0048 0.0051 0.0033 0.0045 0.0050 0.0097 7 6 5 8 8 3 0.0906 0.0990 0.0800 0.0904 0.0928 0.0975 6 1 9 4 1 3 0.8658 0.3417 0.2382 0.2070 0.3996 0.4436 5 6 3 3 5 0.0219 0.0259 0.2768 0.0315 0.0209 0.0313 8 1 3 7 The concentration of essential trace elements in ppm in both treated and control plants 48.432 6.2486 12.837 24 87 153.38 327.54 191.06 33 59 41 24.118 39.726 28.752 42 41 54 117.66 125.57 79.876 29 14 9 The concentration of representative 5.4209 4.7242 10.521 6 4 06 230.64 223.86 347.21 354.64 76 16 56 86 32.274 64.463 22.598 56.512 37 14 208.21 261.43 69.836 92.973 51 21 78 elements in ppm in treated and control plants 1.3313 4 0.0002 8 plants 2.1488 1 0.0022 5 0.1437 2 0.296 0.3196 5 0.0017 1 0.0052 0.0003 2 0.1037 4.095

Na K Mg Ca

4.4918 9 199.91 87 30.564 26 166.75 33

9.9538 9 67.446 2 12.321 66 82.499 49

25.654 94 84.353 68 9.1504 3 52.900 36

Al TI

1.3390 7 0.0002 5

2.5630 1.3561 0.9678 0.6911 0.7559 1.6355 1 1 2 5 7 0.0006 0.0001 0.0001 0.0002 0.0002 0.0001 7 2 4 6 3 3 The concentration of non essential elements in ppm in treated and control 2.3846 2 0.0039 9 0.1805 9 0.0383 3 0.4625 7 0.0052 0.0090 7 0.0007 1 0.0759 1.9788 6 0.0084 6 0.1156 2 0.3376 8 0.3107 5 0.0018 4 0.0015 0.0002 7 0.2258 2.3350 1 0.0075 5 0.1590 3 0.1296 0.2658 4 0.0008 7 0.0046 6 0.0001 6 0.0919 2.0804 3 0.0012 0.1530 1 0.1446 8 0.48 0.0004 7 0.0033 1 0.0001 6 0.3036 2.4293 9 0.0020 5 0.1608 3 0.0914 0.7723 5 0.0004 6 0.0015 9 0.0002 5 0.1608 2.2912 8 0.0017 5 0.1444 3 0.6367 6 0.2153 0.0015 23 0.0013 4 0.0001 7 0.1870

0.5693 6 0.0001 3

0.6411 8 0.0002 1

Si As Se Rb Sr Ag Cd Sb Ba

2.3247 1 0.0018 3 0.1407 3 0.1616 7 0.5743 9 0.0320 7 0.0008 7 0.0002 6 0.4078

2.2319 8 0.0016 3 0.1775 3 0.1514 7 0.2140 4 0.0006 1 0.0013 4 0.0002 3 0.2890

2.2023 5 0.0017 3 0.1554 3 0.0259 5 0.2018 7 0.0003 5 0.0029 0.0002 6 0.2944

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Sr. Prema Kumari, et al.: Comparative Study of Metal Uptake by Medicinal Plants Growing In Natural and Polluted Area of Eastern Ghats

Pb U

1 7 1 7 6 4 0.0126 0.0431 0.0140 0.0271 0.0201 0.0247 0.0199 7 2 4 1 8 2 3 0.0007 0.0017 0.0003 0.0004 0.0002 0.0004 0.0003 3 2 1 7 6 9 6 The concentration of toxic, heavy metals in ppm in treated and control plants

1 0.0323 7 0.0007

9 0.0121 4 0.0002 2

0.0314 1 0.0009 2

Table.2b ; Showing metal concentration in 10 natural and polluted plants

Eleme nt Fe 006 116 007 117 008 118 009 0.7727 6 0.0015 3 0.4678 1 0.0643 1 0.113 0.0033 8 0.0785 4 0.3839 9 0.0224 6 119 2.1658 4 0.0018 5 0.6722 3 0.0916 2 0.0353 0.0065 7 0.0865 5 0.4353 4 0.0270 8 010 1.3892 2 0.0023 7 0.6825 4 0.1432 3 0.0019 3 0.0069 3 0.0838 1 0.4347 3 0.0384 3 120 0.6387 5 0.0007 2 0.5965 9 0.0742 6 0.0025 7 0.0029 3 0.0847 0.1411 0.0168 7 0.8909 4.2936 0.6261 0.1749 0.9675 1.0689 7 8 4 9 Co 0.0049 0.0047 0.001 0.0016 0.0019 0.0023 5 4 2 4 5 Zn 0.3813 0.8085 0.2756 0.4207 0.5024 0.6603 2 5 9 9 Cu 0.0941 0.1281 0.0687 0.1978 0.1327 0.1420 1 7 1 4 7 1 Mo 0.0025 0.0038 0.0012 0.0067 0.0034 0.0016 6 3 9 4 6 V 0.0033 0.0138 0.0028 0.0042 0.0043 0.0068 3 2 4 2 3 3 Cr 0.0818 0.1009 0.8199 0.0893 0.0853 0.0996 8 6 5 5 5 Mn 0.6059 0.6467 0.1802 0.1698 0.1703 0.2676 5 3 2 4 4 2 Ni 0.0232 0.0334 0.2844 0.0215 0.0233 0.0237 2 4 3 The concentration of essential trace elements in ppm in both treated and control plants Na 11.092 5.5944 13.459 56.839 42 4 75 63 K 122.72 83.152 144.10 79.593 02 34 61 73 Mg 33.824 25.772 30.697 15.637 06 27 26 8 Ca 133.74 144.42 236.64 122.97 89.426 83 96 94 58 18 The concentration of representative elements in ppm in treated and control Al 4.3345 1 181.50 15 27.363 55.802 26 120.48 09 15.472 17 84.217 59 plants

4.9609 7 234.12 93 23.809 12 134.99 43

7.0729 1 162.33 11 33.627 19 190.47 75

5.7742 4 223.08 15.418 53 85.058 83

10.177 36 247.79 14 12.733 24 55.856 01

0.9618 1.8782 0.5689 0.7119 1.0654 1.4748 1 1 3 4 5 7 TI 0.0000 0.0002 0.0001 0.0002 0.0000 0.0001 8 6 4 2 8 9 The concentration of non essential elements in ppm in treated and control plants Si 2.2822 6 0.0012 2 0.1449 7 0.2384 5 0.5833 6 0.0005 1 0.0018 1 0.0002 1 1.2874 4 0.0281 7 0.0002 5 109138 0.0056 9 0.1699 8 0.0900 3 0.3376

0.7441 3 0.0000 8

1.2688 8 0.0001 7

1.4686 8 0.0001

0.6201 2 0.0007

2.1359 2.3403 2.2645 2.2917 2.2423 2.5118 1.8668 2.3628 9 3 3 9 7 1 1 8 0.0010 0.0017 0.0013 0.0016 0.0012 0.0030 0.0018 0.0012 1 9 3 7 6 9 Se 0.1422 0.1509 0.1496 0.1821 0.1466 0.1579 0.1496 0.1612 9 7 1 5 1 2 Rb 0.1147 0.0933 0.0902 0.074 0.1618 0.0855 0.1105 0.2351 5 8 8 2 9 Sr 0.8427 0.3857 0.3023 0.3961 0.3858 0.4857 0.2603 0.1645 9 1 4 8 6 8 7 Ag 0.0005 0.0003 0.0004 0.0005 0.0006 0.0006 0.0006 0.0004 0.0004 9 3 2 6 9 4 31 6 6 Cd 0.0025 0.0009 0.0031 0.0009 0.0045 0.0010 0.0020 0.0012 0.0012 9 2 7 7 2 1 3 8 Sb 0.0005 0.0001 0.0002 0.0002 0.0004 0.0001 0.0003 0.0002 0.0003 7 9 6 9 1 6 7 3 4 Ba 12151 0.4417 0.0863 0.5616 0.4467 0.1891 0.1078 0.2400 0.0618 6 2 2 2 9 7 5 3 Pb 0.0361 0.0132 0.0578 0.0144 0.0391 0.0162 0.0258 0.192 0.0244 8 3 3 5 8 8 5 U 0.0012 0.0002 0.0003 0.0003 0.0006 0.0002 0.0006 0.0003 0.0004 2 1 6 2 4 7 4 2 The concentration of toxic, heavy metals in ppm in treated and control plants Control plants: 001,002,003,004,005,006,007,008,009,010. Polluted plants: 111,112,113,114,115,116,117,118,119,120. Abutilon indicum 001: Control 111: Treated Achyranthes aspera 002: Control 112: Treated Adhatoda vasica 003: Control 113: Treated Alternanthera sessilis 004: Control 114: Treated Eucalyptus globules 005: Control 115: Treated Hyptis pectinata 006: Control 116: Treated Azadirachta indica 007: Control 117: Treated Ocimum sanctum 008: Control 118: Treated Recinus communis 009: Control 119: Treated Tinospora cordifolia 010: Control 120: Treated As

The variation in elemental concentration is mainly attributed to the differences in botanical structure, mineral composition of the soil, preferential

absorbability, use of fertilizers, irrigation of water, climatologic conditions. (9) Further the difference is

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Sr. Prema Kumari, et al.: Comparative Study of Metal Uptake by Medicinal Plants Growing In Natural and Polluted Area of Eastern Ghats

sampling andpresence pollution sources can also effect the concentration of 7 metals from species to species. Fe concentration of ferrous compounds is also influenced due to waste released from coromandel fertilizers on which these plants are grown. Iron is a structural component of heme proteins in plants many of which are enzymes of the Krebs cycle(10). In general the concentration of iron in plants from polluted area is higher (26). However in our study out of 10 plants some plants are showing more iron accumulation and few show accumulation in less concentration for unknown reason. Of all the elements that have been analysed in the above study, Na is accumulated in higher concentration in most of the plants 8 out of 10 plants growing in polluted sources and the plants growing in natural sources have significantly low concentration of Na. The role of Na to render the plants to adopt metabolically; to changing metal stresses is yet to be further analysed by studying accumulated metabolites in these plants. Of the 12 elements that are accumulated in higher concentration Na (8 plants) & Co (two plants) are the beneficial elements. In the above plants showing luxuriant growth in polluted sources, the beneficial elements could be compensating for toxic effects of other elements or replacing the mineral nutrients in some other less specific function such as maintenance of osmotic pressure. And also it is surprising to know that percentage of Na in soil of both natural & polluted sources is found to be almost same. And yet the Na uptake by about (8 out of 10 plants) growing in polluted sources is significantly more. The reason could be, the plants could have developed some mechanism such as (Ion flux) which may prevent the plant from the absorption of heavy toxic elements even though they are found in higher concentration in the polluted soil sources. Na & K metal conc are responsible to maintain normal hydration & osmotic pressure and K conc, is needed for cell growth & function (8). Thus the high content of k could be the reason for the luxuriant growth of the medicinal plants under heavy polluted areas. Next to Na, Fe accumulated in higher concentration in the plants grown in polluted metal sources. Fe is the 4th most abundant element in the earths crust. Special iron chelating agents could be available in the polluted; sources that facilitate the solubilization and uptake of Fe (5). Subsequently Pb, Cd & Fe also accumulated in more in plants growing in polluted sources. Higher concentration of Pb, Cd in plants grown in polluted sources might reflect the concentration of these commonly encountered metals in polluted soils, which are being continuously released from the surrounding industries such as Coromandel fertilizers, Hindustan Petrolium Corporation Limited, Hindustan Shipyard, Steel Plant etc. Chelate assisted phyto extraction has been developed because plants do not naturally accumulate important toxic elements eg. Pb, Cd, as that would be significant in remediation: Continuous Phyto extraction of metals relies on properties of plant that lead to accumulation in aerial plant tissues (14, 17).

Ba, Cr, Cu, V, Zn and Fe though present in more concentration in polluted sources, not absorbed much by the plants. Ca & Ti are accumulated in high but not in significant levels. Zn, Co and Mn are essential elements required for various biochemical process and enzymatic process (18, 19). The high concentrations of Mg, Ca and K in the plants represents that these elements are most abundant metal constituents in plants (12). For the higher plants to serve as monitors of metal pollution is dependent upon an understanding of the metabolic processes which enable plants to acquire needed nutrients in tolerate increasing level of toxic elements and analysis in this approach is another process. Trace elements play an important role in the production of Secondary metabolites which are responsible for pharmacological actions of medicinal plants (16). Further it is aimed to study the concentration of these metals that are listed at higher level in the selected medicinal plants and later to intensify the study on their biological effects.

ACKNOWLEDGEMENT
We sincerely thank NGRI (CSIR), Geochemistry division, Hyderabad who supported this research in all the ways possible and we owe to Dr. Varaprasad Bobbarala, Chief Scientist of TRIMS Labs, and Visakhapatnam for guidance and support.

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Source of support: Nil, Conflict of interest: None Declared

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