Temporal Assessment of Crop Protection Agents in Water Bodies of Haryana Districts

Water is one of the most precious natural resources, which is essential for life. The presence of xenobiotics, namely pesticides in water is a matter of global concern. Several pesticides, belonging to different chemical class are being used in agriculture throughout the world Sneha et al., (2012). Pesticides move into water bodies due to wet precipitation Kumar et al., (2007), leaching, field runoff, municipal, agricultural and industrial disposal. In many developing countries water from rivers, canals, wells, tube wells are used both for drinking and irrigation purpose.
       
The contamination of the ground water with several of the organochlorine pesticides (OCPs) has been reported (Singh et al., 2005). Surface water contamination may impact eco-toxicological effect on aquatic flora and fauna and also on human health through food chain Miyamoto et al., (1990). Contamination of ground water resources by pesticides is a matter of grave concern Papadopoulou-Mourkidou et al., (2004). DDT, is under restricted use for disease vector control (UNEP 2003). The major issue arises in the countries where ground water is the source of drinking water for rural and adjacent urban areas (Tuxen et al., 2000). 
       
The water supply in north of India is from the rivers, tributaries in the region. Haryana in northern part is mainly responsible for supply of water to the National Capital, Delhi. Haryana is predominantly an agricultural state in India extends between 27°39'N to 30°55'n latitude and 74°27' to 77°36'E longitude and covers a total geographical area of 442100 ha. The state’s economy is mainly based on agriculture and industry. The major sources of irrigation and drinking water of the region are Anangpur dam, Hathni Kund Barrage, Sutlej Yammuna link canal, Munak Canal, Agra Canal etc. The study highlights the contamination of pesticides in irrigation water bodies like wells, tube wells, canals, ponds and rivers of region from Panipat, Karnal, Rohtak, Sonipat and Hissar districts of Haryana, India.

Geographical location
 
Total 233 water samples collected from the water bodies from cites of Panipat, Karnal, Rohtak, Sonipat and Hissar districts of Haryana, India. The water samples from irrigation system i.e. wells, tube wells, canal and river system were collected throughout the different seasons of the year for the period from 2014-2018. Fig 1 shows districts map of Haryana. Water samples were collected in one-Litre capacity pre-cleaned and rinsed amber-colored bottles. Samples were stored in refrigerator at a temperature of 4°C till analysis to prevent degradation of pesticide residues before processing.
 

 
Na₂SO₄, (AR-grade), sodium chloride (AR grade) dichloromethane, acetone, methanol and n-hexane solvents of pesticide residue grade purchased from Merck (Darmstadt, Germany). 1L separating funnel, 250 ml round bottom flask, volumetric flask of 100, 50, 10 ml was used for standard stock solution and its mixture preparation. 1.5 ml sample vials were used for final sample extract injection to instrument. Turbo-Vap LV evaporator (Caliper Life Sciences, USA), Liquid dispensers (Coleparmer USA), adjustable pipette 1 to 5 ml (Eppendroff, USA) was used.  An electronic weighing balance (Mettler Germany) with digital display was used to weigh the certified reference materials (CRM) and other reagents purchased from Sigma Aldrich, USA whose purity were above 99%.
       
The samples were extracted using conventional liquid-liquid partitioning method for the determination of organic pollutants in water [AOAC Official Methods (2019) and Veeraiah and Prasad (1996)].  The method was validated for its effectiveness. The LOD, LOQ recovery, repeatability was calculated. The samples were processed for residue estimation of different groups of pesticides viz. organochlorines (OC), synthetic pyrethroids (SP) and organophosphates (OP).  The samples were extracted as per above method and were injected in gas chromatography–mass spectroscopy (GC-MS) at Institute of Pesticide Formulation Technology.
       
The water samples were screened for 55 pesticides (Table 1). The Certified Reference Material (CRM) were purchased from Sigma Aldrich (USA). Individual pesticide solution of 100 mg/L concentration was prepared in n-hexane. 55 individual pesticide standard stock solutions were prepared for the preparation of pesticides mixture of 10 mg/L. Working standards and were prepared by serially diluting 10 mg/L pesticides mixture stock as per the sensitivity of the instrument. The standard stock and working solutions were stored at 4°C.
 

       
Gas chromatography system (Agilent Technology 7890A) with DB-5MS fused silica capillary column (Agilent JandW, GC Column, 30 m × 0.25 mm × 0.25 µm) was used for quantitative analysis.  Agilent Chem Station System software was used for instrument control and data analysis. Gases were passed through purification filters which can trap oxygen and moisture, before supply to the column. Agilent mass spectrometer (5975C inert XL EI/CI MSD) was used for identification and quantification and confirmation of 55 pesticides. The mass spectrometer was operated in the positive ion electron impact energy of 70 eV and an emission current 60 μA. Full scan data were obtained with a mass range of m/z 45–500. Scanning interval and SIM sampling rate were kept at 0.5 and 0.2 s, respectively. The full scan TIC chromatogram of Standard Mixture (0.5 ppm) and Control water samples are given in Fig 2 and 3 respectively. The presence of pesticides residues were quantified and confirmed by comparing the retention time, peak area, m/z of the sample with those of the standards. The fragmentation patterns of detected pesticides are given in Table 2.
 

 

 

       
Liquid-liquid partitioning extraction followed by gas chromatographic detection (USEPA 1980; AOAC Official Methods (2019) and Veeraiah and Prasad (1996) was used for the determination of pesticide residues. The percent recovery was performed to evaluate the procedure, by spiking 55 pesticides mixture with known concentrations. For water samples, the mean recovery ranged from 75 to 90%, with a relative standard deviation between 5-20%, at 0.050 μg/L level. The detection limit was 0.010 μg/L. The efûcacy of the methodology was within the 95% conûdence level. The instrument is repatable with the precision (RSDr), precision (RSDwr) is below 5% and LOQ for the pesticides is 0.050 ppm (As per SANTE guideline).

Total 233 water samples were analysed for the presence of 55 pesticides, which were collected from cities of Panipat, Karnal, Rohtak, Sonipat, Hissar and Narnaul districts of Haryana, India (Table 3). Samples were collected during pre and post monsoon and pre and post winter season. Most of the crops are grown during these seasons due to conducive weather conditions. Intensive agriculture during this period leads to high irrigation and frequent use of fertilizers and pesticides. This may lead to runoff from fields into water bodies and aquifers. Temporal assessment of pesticides in water bodies during these months may give an insight into the levels of crop protection agents in water and their safety towards consumption for drinking purpose. The pesticides selected for analysis are based on the consumption pattern of pesticides in the area and the crops grown.
 

       
Analysis of results of Karnal districts indicate that 19 water samples showed the presence of pesticides, out of which multiple pesticide residues were detected in 11 samples. The pesticides detected in the samples were chlorpyrifos, triazofos, fipronil, bifenthrin, pendimethalin, butachlor, cypermethrin, pretilachlor, lambda-cyhalothrin, deltamethrin and buprofezin (Table 4). Chlorpyrifos was detected in 12, butachlor in 5, pretilachlor in 7 and lambda cyhalothrin in 7 samples. Water samples along the 230 km stretch of the riverin Haryana were analyzed for  the  presence  of organochlorine insecticide residues (Kaushik et al., 2010). Among 100 farmers, a survey was conducted in cauliflower and tomato cultivating areas of district Faridabad, Haryana, India from December 2012 to February 2013. Cypermethrin (62%) and profenofos (58%) were found as the most popular insecticides (Tyagi et al., 2015).
 

       
The range of contamination of pesticides in water samples was 2-1500 µg/L. Total 45 samples were collected from Rohtak in which 7 samples showed the presence of pesticides. Butachlor was detected in 7 samples (Table 4). From 30 and 15, samples from Sonipat and Narnaul, one sample each showed the presence of pesticides, (Table 3). A total 233 water samples were analyzed; out of which 70 samples were from Panipat, 43 samples from Karnal, 45 samples from Rohtak, 30 water samples from Sonipat, 26 samples from Hissar and 15 samples from Narnaul. Amongst the samples analyzed, 25 water samples showed the presence of at least one pesticide and in 11 samples multiple pesticide residues were recorded. Chlorpyrifos, butachlor, pretilachlor and cypermethrin were detected in 14, 12, 8 and 7 samples, respectively, whereas 2 samples showed the presence of triazofos and cypermethrin and 1 sample recorded the presence of five pesticides namely, fipronil, bifenthrin, pendimethalin, deltamethrin and buprofezin respectively (Table 3 and 5).
 

       
Temporal distribution of pesticides in water samples during different seasons is presented in Table 5. The pesticides detected during seasonal variation in 68 water samples of monsoon (pre and post) and 165 samples of winter (pre and post). It is observed that about 70% of crops are cultivated during winter season from mid-October to mid-April whereas only 30% of crops are grown during rainy season and localized farming is done during the months June to October. Thus, leading to increased inputs during winter season and subsequently prompting collection of larger number of water samples in winter season than during the monsoon. A total of 68 samples were collected from Panipat, Rohtak, Karnal, Hissar and Sonipat during monsoon seasons (pre and post), among which 16 samples showed the presence of pesticides residues and 10 samples recorded the presence of more than one pesticide. Among the 165 samples of winter season, only 11 samples recorded pesticides and multiple pesticide residues were detected in 3 samples. The low pest incidence during the winter season leads to lesser use of pesticides and resulting in lower detection of pesticides in water samples during the winter months. In contrast, the pest and disease attack during the monsoon season is high due to the conducive climatic conditions, thus prompting high use of crop protection agents and leading to higher detection of pesticides. Regulatory limits by various regulatory bodies (Hamilton et al., 2003) has been compared with the contamination of the detected pesticides. The water samples of Panipat and Hissar region detected pesticide levels below the quantification level (Table 4 and 1). Chlorpyrifos was detected in the range of 1-1515 µg/L, BQL (<0.050 µg/L) -30 µg/L and BQL (<0.050 µg/L)-1 µg/L in Karnal, Sonipat and Narnaul samples, respectively. The level of concentration in 13 water samples of Karnal districts were above EPA 8081 and 8141 value of 0.3 µg/L but 4 samples were below UAS health advisory life time value of 20 µg/L. Chlorpyrifos detected in one samples from Sonipat was above EPA and USA health advisory life time, however one sample of Narnaul was above EPA but below USA Health advisory life time. Triazofos, pretilachlor, bifenthrin, fipronil and buprofezin were detected in the range of 122-439 µg/L; 87-906 µg/L, BQL (<0.050 µg/L-33µg/L; BQL (<0.050 µg/L-18 µg/L and BQL (<0.050 µg/L-2 µg/L, respectively in Karnal water samples, no limit has been ascertained by any of the regulatory authorities. Butachlor pesticide detected in Karnal and Rohtak water samples ranged from 16-51 µg/L and 0.2-28 µg/L, respectively. Seven water samples from Karnal recorded the presence of lambda cyhalothrin in the range of 5-520 µg/L and were above the EPA value of 0.05 µg/L. Similarly, cypermethrin residues in 2 water samples of Karnal and 1 sample of Rohtak was in the range of 728-935 µg/L and 60 µg/L, respectively were above EPA value of 0.25 µg/L. In one sample from Karnal, deltamethrin was detected at 292 µg/L concentration which was above EPA value of 0.25 µg/L. A similar study conducted on groundwater samples from tubewells during the year 2002-2003 in Hissar revealed that synthetic pyrethroids like deltamethrin was detected in water samples varying from  0.017-0.06 µg/L and cypermethrin  from 0.022-0.090 µg/L. The most predominantly detected pesticide was chlorpyrifos, in the range 0.607-1.121 µg/L, (Kumari et al., 2008). Chlorpyrifos was detected from groundwater samples of the agricultural area of Amravati, Bhandara and Yavatmal region of Maharashtra in the concentration of 0.25, 0.11 and 0.18 µg/L, respectively (Lari et al., 2014). The studies conducted by Kumar et al., (2007), in Hissar district of Haryana revealed that the rain water were detected with cypermethrin (0.2-1.0 µg/L), deltamethrin (0.1-0.8 µg/L) and triazofos (0.05-0.009 µg/L) pesticides.

Panipat, Karnal, Rohtak, Sonipat, Hissar and Narnaul are the prominent agricultural districts of Haryana, India. Sufficient numbers of samples were collected for analysis of pesticides from these districts of Haryana. Temporal and seasonal pesticide residue variation was focused in the study. Pesticide residues in the water samples from Panipat and Hissar regions were below the quantification levels. 40.4; 15.5; 0.03 and 0.07% of pesticides were present in Karnal, Rohtak, Sonipat and Narnaul samples, respectively. In 23.4 and 0.04% of water samples of Karnal and Rohtak multi pesticide residues were detected Temporal variation reveals that 23.5% and 6.6% of pesticides were recorded in 68 and 165 samples of monsoon and winter samples. 0.15 and 0.02% of water samples from Karnal and Rohtak area showed the presence of multiple pesticide residues Chlorpyrifos, pretilachlor, butachlor and lambda chalothrin were predominantly present in all water samples analyzed. Water samples from Karnal detected with chlorpyrifos were above EPA method 8081 B and 8141 but few samples were below UAS Health advisory life time. Some water samples from Karnal region, detected with chlorpyrifos, deltamethrin, lambda cyhalothrin were above EPA method 8081 B and 8141. One sample each from Sonipat and Narnaul showed the presence of chlorpyrifos above EPA method 8081B and 8141. Cypermethrin residues in water sample from Karnal and Rohtak were above EPA method 8081B and 8141 value.

The authors are highly thankful to Directorate of Horticulture, Haryana and Director IPFT for their help and support.

This work was supported by Institute of Pesticide Formulation Technology.

No potential conflict of interest was reported by the author.