INTRODUCTION:

Phytoremediation:

Phytoremediationis a technique for remediating polluted soils through the use of plants. Pollutants can be both inorganic and organic chemicals. In several European countries, biological clean-up techniques are finding interest and are supported by Governments and research organizations. Phytoremediation has several benefits: It is inexpensive, it seems to be effective, it is in situ, and it is 'green' (Jackson, D. R.;et, al., J. Environ. Qual. 1977). A special advantage of phytoremediation compared to other techniques is that the soil functioning is maintained and life in soil is reactivated. Human evolution has led to immense scientific and technological progress. Global development, however, raises new challenges, especially in the field of environmental protection and conservation (Bennett et al., 2003). Nearly every Government around the world advocates for an environment free from harmful contamination for their citizens. However, the demand for a country’s economic, agricultural and industrial development outweighs the demand for a safe, pure, and natural environmental. Ironically, it is the economic, agricultural and industrial developments that are often linked to polluting the environment (Pandey AK. National Academy Science Letters, 2007).

Parthenium Hysterophorus L. (Asteraceae), also known as congress grass, is an annual herb, invasive weed throughout India and world. It is an aggressive colonizer of wastelands, pastures and road sides in India. The leaf proteins are reported to be better than cereal and legume proteins. It is used as spices in many parts of the world. Parthenin free dried fibers of plants are used as cattle feed [Narasimhan TR et.al; 1993]. All parts of the plant are reported to be used as bitter tonic, febrifuge, emmenagogue, antidysenteric [Oudhia P.; 2001]. It was found much traditional method for treatment of fertility, diabetes mellitus, cancer and many more disorders.

Lead:

Lead is one of the major heavy metals contaminations in soil, which has gained negative impact on environmental pollutant. It results from mining, paints, dyes, gasoline and explosives as well as disposal of municipal sewage sludge. So Lead contamination increase in soil, leads to soil toxicity (Yang et, al 2000). These hazardous will be uptaken by the plant, which leads plant product hazardous that is to harm human health. It enters by intake of plant product, which accumulates in body organ like brain. It may cause death (Pallavi sharma, et al.; Department of Biochemistry, BHU, 2005).

MATERIALS AND METHODS:

Collection of Plant Material and Authentication:

Fresh whole plant of Parthenium hysterophorus linn were collected from the Kelambakkam, Kanchipuram Dist.The collected plant was identified and authenticated by Dr. D. Aravind Assistant Professor, Department of Medicinal Botany,National Institute of Siddha,An Autonomous body under the Ministry of AYUSH, Govt. of India, Tambaram, Sanatorium, Chennai-600047, Tamil Nadu, India vide authentication certificate number NISMB2182O16 dated on 1.3.2016.

Collection and addition of heavy metals to soil :

Plantation of P. Hysterophorus linn seed to soil which has different concentration of lead to check phytoextraction of lead by plant .

Detection of Heavy Metals in the Soil by AAS:

The suitable amount of the soil samples in evaporating dishes were taken and acidified to methyl orange with conc.HNO₃. Further, 5ml conc. Nitric acid was added and evaporated to 10ml. Then it was transferred to a 125ml conical flask. 5ml of conc. Nitric acid and 10ml of perchloricacid (70%) were added. Then heated gently, till white dense fumes of HClO4 appear. The digested samples were cooled at room temperature, filtered through whatmann no41 or sintered glass crucible and finally the volume was made upto 100ml with distilled water. Then this solution was boiled to expel oxides of nitrogen and chlorine.This solution contained 0.8N inHClO4.The solution was used for the use of determination of heavy metals.

PROCEDURE:

Apparatus Set-up:

Inlet of reaction cell was connected with auxillary purging gas controlled by flow water. Incase of need of a dry IMF cell between the reaction cell and atomizer, only anhydrous CaCl₂was used. Before using the hydride generation, operating parameters were optimized. Dilute aqueous solution of as was aspirated directly into the flame to facilitate atomizer alignment. Quartz atomizer for max. Absorbance was aligned. A blank was aspirated until memory effects were removed. Recommended wavelength is 22nm for as determination. 0,1,2,5,10,15and 20 mg standard solutions of Pb( III) were transferred to 100ml volumetric flask and made-up to the volume with soil containing the same conc.of 50% v/v acid (40% HCl and10% H2SO4) used for sample preservation (commonly 2-5ml) conHNO3/L). This yielded blank and standard solutions of 0, 1,2,5,10,15 and 20 mg Pb/L. Preparation of samples and standards for total 50 ml sample or standard was added to a 200mL Berzeliusbeaker and then added 1ml, 2.5N H₂SO₄and 5ml K₂S₂O₈. Then the solution was gently boiled on pre-heated hotplate until a final volume of 10 ml was reached. After Digestion, it was diluted to 50 ml for subsequent as measurement.

Statistical Analysis:

Experiment was conducted in triplicate (N=3). Results are shown as mean () standard error. Experimental data were analysed using statistical software SPSS 16.0(M. Krishnaveni., 2012).

RESULT :

Soil Study Report:

Collected soil sample was characterized by Atomic Absorption Spectrophotometer (AAS) for the presence of heavy metals such as Cd, Pb, as and Hg. Background concentration of lead in the soil was 28.91 mg of lead /kg of soil and others were below detection level, was shown in table- {1-4}. Seeds of Parthenium hysterophorus linn. were grown into soil sample containing 200mg of lead /kg of soil, 400 mg of lead /kg of soil, 600 mg of lead /kg of soil, 800 mg of lead /kg of soil and 1000 mg of lead /kg of soil respectively in pots. After 60 days, soil was characterized by AAS. Percentage removal of lead mg/kg of soil were 17.8%, 28.87%, 37.3%, 19.6% and 14.35% respectively. This was shown in fig-2 and table {6-7}.

Fig. 1: shows Parthenium Hysterophorus Linn.seeds Germinated and Its growth (60 days).

Table-1: AAS Report of lead (Pb) toxicity in soil sample before germination of Parthenium hysterophorus linn. Seeds.

Sl. No.	Sample ID	Analyte Lead(Pb) nm	Mean (conc. mg/L)	Std. Dev	% RSD
1.	Calib Blank 1	283.31	0.00	0.00	34.66
2.	Calib Std 1	283.31	0.2 mg/L	0.00	0.70
3.	Calib Std 2	283.31	0.5 mg/L	0.00	10.46
4.	Calib Std 3	283.31	1.0 mg/L	0.00	1.00
5.	Calib Std 4	283.31	2.0 mg/L	0.00	0.51
6.	Calib Std 5	283.31	3.0 mg/L	0.00	1.00
7.	Calib Std 6	283.31	10.0 mg/L	0.00	1.00
8.	Calib Std 7	283.31	20.0 mg/L	0.00	0.82
9.	Blank	283.31	-0.1134 mg/L	0.00696	6.14
10.	ICV-1mg/l	283.31	1.015 mg/L	0.0417	4.11
11.	M-3258/15-16	283.31	0.2919 mg/L	0.00945	3.24
12.	CCV-1 mg/l	283.31	1.081 mg/L	0.0207	1.92

Table- 2: AAS Report of cadmium (Cd) toxicity in soil sample before germination of Parthenium hysterophorus linn. Seeds.

Sl. No.	Sample ID	Analyte Cadmium (Cd) nm	Mean (conc. mg/L)	Std. Dev	% RSD
1.	Calib Blank 1	228.80	0.00	0.00	44.55
2.	Calib Std 1	228.80	0.2 mg/L	0.00	16.64
3.	Calib Std 2	228.80	0.5 mg/L	0.00	0.04
4.	Calib Std 3	228.80	1.0 mg/L	0.00	2.31
5.	Calib Std 4	228.80	2.0 mg/L	0.00	3.29
6.	Calib Std 5	228.80	3.0 mg/L	0.00	0.12
7.	Calib Std 6	228.80	10.0 mg/L	0.00	1.32
8.	Calib Std 7	228.80	20.0 mg/L	0.00	1.76
9.	Blank	228.80	0.0056mg/L	0.00777	139.57
10.	ICV-0.1mg/l	228.80	0.1066mg/L	0.00383	3.59
11.	M-3258/15-16	228.80	0.0041mg/L	0.00089	21.78
12.	CCV-0.1 mg/l	228.80	0.1061mg/L	0.00262	2.47

Table-3: AAS Report of arsenic (As) toxicity in soil sample before germination of Parthenium hysterophorus linn. Seeds.

Sl. No.	Sample ID	Analyte Arsenic (As) nm	Mean (conc.) microgram/Litre	Std Dev	% RSD
1.	Calib Blank 1	193.70	0.00	0.00	8.32
2.	Calib Std 1	193.70	1.0 mg/L	0.00	5.84
3.	Calib Std 2	193.70	2.5 mg/L	0.00	4.36
4.	Calib Std 3	193.70	5.0 mg/L	0.00	0.72
5.	Calib Std 4	193.70	10.0 mg/L	0.00	3.20
6.	Calib Std 5	193.70	25.0 mg/L	0.00	0.02
7.	Calib Std 6	193.70	50.0 mg/L	0.00	0.45
8.	Calib Std 7	193.70	100.0 mg/L	0.00	0.43
9.	Blank	193.70	-0.2813mg/L	0.11332	40.29
10.	ICV-0.05mg/l	193.70	4.646mg/L	0.1484	3.19
11.	M-3258/15-16	193.70	0.2758mg/L	0.06650	24.11
12.	CCV-0.05 mg/l	193.70	4.858mg/L	0.2877	5.92

Table-4: AAS Report of mercury (Hg) toxicity in soil sample before germination of Parthenium hysterophorus linn. Seeds.

Sl. No.	Sample id	Anylate Mercury (hg) nm	Mean (conc. µg /L)	Std dev	% RSD
1.	Calib Blank 1	253.65	0.00	0.00	80.86
2.	Calib Std 1	253.65	0.001 µg /L	0.00	10.30
3.	Calib Std 2	253.65	0.005 µg /L	0.00	0.14
4.	Calib Std 3	253.65	0.01 µg /L	0.00	0.04
5.	Calib Std 4	253.65	0.02 µg /L	0.00	0.30
6.	Calib Std 5	253.65	0.03 µg /L	0.00	0.08
7.	Calib Std 6	253.65	0.04 µg /L	0.00	0.04
8.	Calib Std 7	253.65	0.05 µg /L	0.00	0.04
9.	Blank	253.65	-0.0007 µg /L	0.00002	2.32
10.	ICV-0.01mg/l	253.65	0.0099 µg /L	0.0000	0.05
11.	M-3258/15-16	253.65	-0.0003 µg /L	0.00001	4.38
12.	CCV-0.01 mg/l	253.65	0.0097 µg /L	0.00000	0.04

Table-5: AAS Test report of given soil sample before germination of Parthenium hysterophorus linn. Seeds.

S. No.	Parameter	Units	Results
1.	Lead (Pb)	mg/kg	28.91
2.	Cadmium (Cd)	mg/kg	BDL (DL:0.2)
3.	Arsenic (As)	mg/kg	BDL (DL:0.2)
4.	Mercury (Hg)	mg/kg	BDL (DL:0.2)

Table-6 : Effect of Parthenium hysterophorus linn. to lead contaminated soil

S. No.	Concentration of Lead (mg/kg of soil )	Concentration of Lead (Pb) after remediation	Reduction of lead (mg/kg of soil )
1.	200	164.4±0.01	35.6
2.	400	284.5±0.05	115.5
3.	600	376.2±0.02	223.4
4.	800	643.2±0.06	156.8
5.	1000	856.5±0.04	143.5

Table-07 : Concentration of lead before and after phytoremediation

Metal	Metal Concentration In Soil (mg/kg)			% Reduction of lead in soil after maturation
Metal	Background concentration	concentration added in the soil	Total concentration	% Reduction of lead in soil after maturation
Lead	28.91	171.09	200	17.8
	28.91	371.09	400	28.87
	28.91	571.09	600	37.3
	28.91	771.09	800	19.6
	28.91	971.09	1000	14.35

Graph-1: Phytoremediation of lead by P. hysterophorus Linn.

Graph-2: % Reduction of lead in soil by P. hysterophorus Linn.

DISCUSSION:

Contamination of soil by heavy metals is of wide spread occurrence as a result of human, agriculture and industrial activities. The contamination is a risk to all organism including human. Entrance to the toxicity metals into the human body beyond threshold limits, causes many disease and health abnormalities(Marimutthu Krishnaveni, Periyar University;2014). Among those hazardous heavy metals lead is a potential pollutant that readily accumulates in soil and sediments. Thus, remediation of heavy metals pollutants in the soil deserve due attention(N. Kumar.,et.al.,1995). Phytoremediation is a better solution to this problem. Phytoremediation is technique with use of plant to reduce the concentration of toxic effect of contaminants in the environment(Harborne JB, New York,1984). Although lead is not an essential element for the plants, it gets absorberd and accumulated in different parts of the plant. Parthenium hysterophorus linn. is a weed, distributed through out the tropics of the world and plains in India(Krishnavignesh Lakshmanan et .al.,2012). The present work established that Parthenium hysterophorus linn. Contains most of the phytoconstituents when tested qualitatively. The methanol extract of parthenium hysterophorus linn was found to be rich in phytocompounds such as polysaccharides, proteins, aminoacids, saponins, tannins, terpenes and phenolics. Hence, it indicates that chemical compounds present in the plant are involved in the removal of lead (Pb) from the soil (Bennett LE,et.,al2003). Emerging cleans up technologies for phytoremediation of lead (Pb) as phytoextraction using Parthenium hysterophorus linn. having the potential to provide environmentally sound and economically viable remedies for the cleaning of lead contaminated soils(Ensley BD, 2000). Further study can be carried out to genetically modified Parthenium hysterophorus linn. For better phytoremediation of lead and molecular studies greatly help in understanding the mechanism and enhancing the efficiency of phytoremedation of Parthenium hysterophorus linn. Furthermore, advance research can be achieved by isolating the active principle from Parthenium hysterophorus linn, involved in great potential for cleans up soil contaminated with lead.

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Received on 03.03.2017 Modified on 30.03.2017

Res. J. Pharmacognosy and Phytochem. 2017; 9(2): 64-68.

DOI: 10.5958/0975-4385.2017.00011.5