Evaluation and assessment of risk of pesticide residue in vegetables, soil and water samples of few district of Northern India

doi:10.21203/rs.3.rs-3808776/v1

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Evaluation and assessment of risk of pesticide residue in vegetables, soil and water samples of few district of Northern India

https://doi.org/10.21203/rs.3.rs-3808776/v1

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The main motive of the study was to determine pesticides residues in 16 different vegetables grown in four districts i.e Rohtak, Karnal, Hisar, Panipat of Haryana, India. In total 102 vegetables of field and market for the pesticide residue analysis during the year 2019-2020. Also along with that 46 soil and 50 water samples were a collected from the same four districts. 161 pesticideswere screened for their presence of pesticide residue. Analysis of samples were done using liquid chromatography–tandem mass spectrometry (LC-MS/MS) (90 pesticides) and gas chromatography–tandem mass spectrometry (GC-MS/MS) (71). The extraction of a multi-residue method using QuEChERS method for vegetables and traditional partitioning and shaking method for water and soil respectively. The evaluation of detected residues where compared with the maximum residue limit (MRL) of FSSAI and European regulations. A total of 31 samples (30.39%) of vegetables, 44 samples (95.65%) of soil and 34 samples (68.0%) of water were detected with pesticide residue. Alpha cypermethrin, hexaconazole and carbendizim were detected in low concentration below MRL and chlorpyrifos, cypermethrin, pendimethalin, buprofezin, butachlor and deltamethrin were deteted in higher concentration in some samples. Chlorpyrifos, cypermethrin, pendimethlin and butachlor were commonly detected. As the result of the study, it is necessary that the government need to support the the monitoring of pesticide residues in vegetables and other environmental samples for consumers’ health.

Vegetables

Soil

Water

Pesticide Residue

Haryana

Haryana plays major role for green revolution for food production. The agriculture sector’s production and productivity of the state has been increased. To improve the livelihood of farmers, the agriculture sector is based on technology whose focus on increased food production. Haryana is one of the 11th states of the country for vegetables production. Haryana is one of the 15th largest population states of farmers which are engaged in agriculture. Of the total 44.96 per cent population are engaged in agriculture (https// www.icfa.org ). In India annual estimated loss of crop due to pests are about US$ 42.66 million (Sushil, 2016). Agrochemicals used are not totally reached the target but maximum are lost to the surrounding environment. The presence of residue in food can affect human health and environment (Abhilash and Singh, 2009). The agrochemicals leads to genotoxic, carcinogenic, respiratory, neurological and mutagenic disruption (Hayat et al., 2011; Choi et al., 2004; Galloway et al., 2003). From the start of the Green Revolution, around 800,000 people died due to pesticides. Around 20,000 people lost their life due to use of pesticide every year for the production of food (Bhardwaj and Sharma, 2013). The Acceptable daily intake (ADI), estimated daily intake (EDI) and maximum residue limit (MRL) represents the maximum concentration of a pesticide residue (mg/kg) and Hazardous risk index (HRI) are the Health safety limits for human health. Pesticides exposure to humans are calculated on the average consumption per person per day, average adult weight, and pesticide residue data (FAO/WHO 2004; Seo et al., 2013).

The study is employed with sensitive, accurate and selective multi-residue methods for the quantitative and qualitative determination of several classes of pesticides of different chemicals and physical properties by GC-MS/MS and LC-MS/MS. The validated analytical method for analyzing pesticides from environmental matrices of vegetables, water, and the soil and the health risk due to the presence of pesticides in a wide range of vegetables are evaluated as per the existing regulations.

The pesticide residue in vegetables, soil and water samples were analyzed in Analytical Division at Institute of Pesticide Formulation Technology (IPFT), Gurugram under the Department of Chemicals and Petrochemicals of Ministry of Chemicals and Fertilizers, Govt. of India, India.

Study area

The study was commenced in Haryana state which is geographically located at 30.73° N and 76.78° E; and having very fertile called the Green Land of India. The collections of samples were done from Rohatk, Panipat, Karnal and Hisar, districts of Haryana. The selection of these districts as it consumed highest amount of agrochemicals in entire state (Fig. 1).

Collection and preparation of sample

Samples of vegetables (102), water (50) and soil (46) were collected from farmer’s farm and local vegetable markets of study areas, during July 2019 to March 2020 by Haryana Horticulture. The amount of each vegetable sample was 1kg, soil was 500gm and 1lit water sample. The collection of samples were done in a clean transparent airtight polyethylene bag/bottle to avoid cross-contamination and kept in cool temperature during transport. The collected samples were transported to the Institute of Pesticide Formulation Technology (IPFT), Gurugram. The analytical lab of IPFT is an accredited laboratory (NABL as per ISO IEC 17025/2017) for pesticide residue analysis. Each sample was homogenised and mixed properly. All the samples were refrigerated at -20 (± 5) °C until extraction and sample preparation.

Chemicals and reagents

Pesticide standards were procured from Sigma-Aldrich and Accustandard Inc. (USA) with certified purities and uncertainty value mentioned on it. The monitored pesticides, their classification and LC-MS/MS and GC-MS/MS pesticides technical data are listed (Wood, 2018; Pesticide Properties Data base A to Z, 2018). Solvents includes Acetonitrile, n-Hexane, Acetone, Methanol and Dichloromethane were HPLC-grade procured from Fischer Scientific and other reagents used for the extraction were anhydrous Magnesium sulphate and sodium sulphate (Merck Life Science, India), sodium chloride (NaCl), Primary Secondary Amine (PSA), C18 and graphite carbon black (GCB) were procured from Agilent Technologies ( USA).

Pesticide standard solution preparation

The stock solution was prepared from the Certified Reference Material (CRM) of each pesticide having specified purity. Weighted CRM of individual pesticide into 10 ml volumetric flask on an analytical precision balance (Sartorius) and make-up to the mark in volumetric flask. The concentration were about 100 mg/Kg which were stored at -20°C. A standard solution mixture for GCMSMS and LCMSMS analysis containing all the aforementioned pesticides in 10 ppm standard mixture of separate 71 and 90 pesticides were prepared respectively. Intermediate standard of 1ppm were prepared from standard mixture of 71 and 90 pesticides separately. Working standard solution at 10, 50, 100, 150, 200 and 250 µg/kg level were prepared for calibration. Procedural standards are prepared by spiking the control matrix sample at 10, 50, 100, 150, 200 and 250 µg/kg level with CRM solutions, prior to extraction. The samples are analysed as per calibration curve of procedural standards.

Plotting calibration curve

The dilutions (10, 50, 100, 150, 200 and 250 µg/kg) of mixture of standard solution of each pesticide group were prepared and injected at specified instrumental method conditions. The samples were analysed (retention time, peak area etc.) against 10, 50, 100, 150, 200 and 250 µg/kg level calibration curve. Each peak was identified by its retention time and mass fragment.

Extraction and cleanup procedure

Preparation of vegetable samples

QuEChERS technique was used for the extraction and clean-up of samples (Anastassiades et al., 2003). Samples were homogenized in a mixer grinder. 10 gram of homogenized sample was weighted in a 50 mL polypropylene centrifuge tube adding in it 10 mL of acetonitrile (MeCN) and shaken for 30 s in vortex mixer. Later added 4 gram anhydrous MgSO₄ and 1 gram of NaCl and shaken quickly for 1 minutes in vortex mixer. Afterwards, the sample tubes were centrifuged at 6,000 rpm for about 5 minutes. For removal of highly pigmented food samples cleanup was performed according to Lehotay et al., 2007. 1.5 mL of aliquot layer was taken into 15 mL micro-centrifuge tube and added with 150 mg anhydrous MgSO₄, 40 mg Primary Secondary Amine ( PSA) and 40 mg C18 thoroughly mixed by vortex for 1 minute and centrifuged for 5 min at 4000 rpm. 1 mL of supernatant was filtered by a 0.45mm PTFE filter and obtained extract were evaporated to dryness under mild nitrogen and reconstituted in 1 mL n-hexane for GC-MS/MS and 1 mL ACN: Water (60:40, v/v) for the LC-MS/MS analysis.

Preparation of soil samples

Homogeneously mixed soil samples were taken (Mishra et al., 2019) and weighed 20 g in 250 mL round bottle flask and added 50 mL of methanol (MeOH) to each flask which was subjected to auto-shaker for 1 hour at 160 rpm. The samples were left undisturbed on flat surface for half an hour for settling soil particles. 20 mL of supernatant was pipette out and filtered through anhydrous sodium sulphate (Na₂SO₄) into 50 mL flat bottom round flask. Also Na₂SO₄ bed was again washed by 10 mL of methanol into same round bottom flask. The collected supernatant solvent was evaporated for dryness under vacuum (500 mm of Hg) at constant temperature of water bath at 50°C. Final volume was reconstituted with 1ml of n-hexane for instrumental analysis using GC-MS/MS and LC-MS/MS.

Preparation of water samples

Collected samples were filtered to remove debris (Mishra et al., 2016). 500 mL of water sample was taken into 1000 mL separating funnel. Then add 100 mL of saturated (NaCl) brine solution. The water sample was extracted thrice with (100:50:50) mL dichloromethane: hexane (80:20) mix; shaken for 4–5 minutes each time with periodic venting. The DCM (organic phase) extract was collected in flat bottom round flask by passing through anhydrous sodium sulphate (Na₂SO₄) for absorbing the moisture present in solvent. The collected solvent was dried in a vacuum rotary evaporator. Later the sample was reconstituted with 5 mL n-Hexane for analysis in GC-MS/MS and LC-MS/MS.

Instrumental Parameters

LC-MS/MS

The samples was analysed using liquid chromatography (Thermo Scientific Accela 600 pump) with a triple quadruple mass detector with electrospray ionization (ESI) source (Thermo Scientific, TSQ Quantum Access) and a Thermo Scientific Hypersil Gold aQ column (50 x 2.1mm, 1.9µm particle size, C-18). The sheath gas flow rate was 48 units, the auxillary gas (AUX) flow rate was 15 units, collision gas pressure 1.5 mTorr the capillary temperature and vaporizer temperature were 320°C and 350°C, respectively, the spray voltage was positive polarity- 4500 V and negative polarity- 3500, the bar Pressure stability was 10, and the cycle time was 1.200 s. Mobile phase A- Aquas ( 0.1% formic acid and 5mM ammonium formate) and mobile phase B- Methanol (0.1% formic acid and 5mM ammonium formate ) were used for the gradient program, with 2% B over 0.40 min and sharply increased to 40% B over 2 min, then linear increased to 95% B over 20 min, and maintained 95% B for 2 min. The column was then reconditioned to 2% B for 3 min. The column’s temperature was set at 25°C. The injection volume was 10 µL at a flow rate of 300 µL/min. Total run time of acquisition was 25 minutes. MRM Transitions of pesticides selected for study for LCMSMS analysis are as follows

1. Mepiquat chloride	2. Thiobencarb	3. Thiodicarb
4. Chlormequat chloride	5. Oxycarboxine	6. Chlorfenviphos
7. Methamidophos	8. Phorate sulphoxide	9. Etoxazole
10. Ethephon	11. S-metolachlor	12. Meptyl dinocap
13. Diquat dibromide monohydrate	14. Myclobutanil	15. Flufenacet
16. Acephate	17. Imazethypyr	18. Phosalone
19. Paraquat dichloride hydride	20. Thiomethoxam	21. Kasugamycin hcl
22. Benomyl	23. Triadimenol	24. Carbosulfan
25. Cymoxanil	26. Bifenazate	27. Flupicolide
28. Simazine	29. Allethrin	30. Diafenthiuron
31. Dinotefuran	32. Fenazaquine	33. Difenthiuron
34. Dodin	35. Tebuconazole	36. Dimethomorph
37. Flonicamid	38. Diflubenzuron	39. Ethiprole
40. Diuron	41. Pretilachlor	42. Mandipropamid
43. Carboxine	44. Hexaconazole	45. Ethoxysulfuron
46. Carbofuran-3-hydroxy	47. Triazophos	48. Triasulfuron
49. Linuron	50. Flusilazole	51. Azoxy strobin
52. Clothianidin	53. Phenthoate	54. Difenconazole
55. Triadimefon	56. Iprovalicarb	57. Triafloxystrobin
58. Fenobucarb	59. Cyzophamid	60. Fenpyriproxyfen
61. Propoxur	62. Benalaxyl-m	63. Cypermethrin
64. Carbofuran	65. Iprodione	66. Temephos
67. Acetamiprid	68. Bitertanol	69. Mpp-dihydrochloride hydrate
70. Monocrotophos	71. Thiophanate	72. Cyantraniliprole
73. Bendiocarb	74. Oxadiazon	75. Chlorantraniliprole
76. Thiacloprid	77. Clodinofop propargyl	78. Flufenoxuron
79. Imidacloprid	80. Hexylthiazox	81. Iodosulfuron methyl
82. Indoxacarb	83. Azadiractin + adduct	84. Abamectin
85. Flubendaimide	86. Spinosad d	87. Chlofenopyr
88. Spinosad a	89. Emamectin benzoate	90. Carbofuran

GC-MS/MS

Shimadzu GCMS/MS system (QP8040) with split/splitless injector were used for analysis. The capillary column was DB-5MS (Agilent J&W GC column, 5% Phenylated methyl siloxane, length: 30 m, I.D.: 0.25 mm, film thickness: 0.25 µm). 1.0 µL of sample volume was injected in splitless injection mode with the injection port was set at 250°C and 1mL/min flow was used for the Helium carrier gas (grade-1). The oven temperature program was initially set to hold at 50°C for 2 minutes then ramp with 1 minutes hold to 190°C at 15°C/min, and finally ramp to 280°C with 10 min hold at 5°C /min. The oven program’s total length was 42 minutes. The interface temperature, ion source temperature and quadruple temperature were set at 280°C and 230°C and 150°Crespectively. The mass spectrometer was operated in electron impact (70ev), selected ion monitoring (SIM/MRM) mode with solvent delay time 5 min. GCMS/MS solution software was used for instrument control and data processing. For the determination of pesticides, the NIST library was used.

Pesticides selected for study for GCMSMS analysis are as follows

1. Propoxur	2. Transfluthrin	3. o,p'-DDD
4. 3,4-Dichloranilline	5. Metalaxyl (Mefenoxam)	6. Buprofezin
7. alpha-BHC	8. Pirimiphos-methyl	9. Kresoxim-methyl
10. Pencycuron	11. Linuron	12. Chlorfenapyr
13. Phorate	14. Malathion	15. Ethion
16. Thiometon	17. Metolachlor (S-Metolachlor)	18. Triazophos
19. Dimethoate	20. Chlorpyrifos	21. Benalaxyl
22. Dazomet	23. Fenthion	24. Propiconazole-1 / Propiconazole-2
25. Simazine	26. Flufenacet	27. Endosulfansulfate
28. Atrazine	29. Pendimethalin	30. p,p'-DDT
31. beta-BHC	32. Fipronil	33. Diclofop-methyl
34. Propetamphos	35. Heptachlor-exo-epoxide	36. Propargite-1 / Propargite-2
37. delta-BHC	38. Chlorfenvinfos	39. Etoxazole
40. beta-BHC / gamma-BHC (Lindane)	41. Captan	42. Fenpropathrin
43. Chlorothalonil	44. o,p'-DDE	45. Fenazaquin
46. Paraoxon Methyl	47. Butachlor	48. Phosalone
49. Etrimfos	50. Endosulfan-I	51. Pyriproxyfen
52. gamma-BHC (Lindane)	53. Hexaconazole	54. L-Cyhalothrin
55. Iprobenfos	56. Fipronilsulfone	57. Permethrin-1 / Permethrin-2
58. Propanil	59. Profenofos	60. Cyfluthrin-1 / Cyfluthrin-2 / Cyfluthrin-3 / Cyfluthrin-4
61. Metribuzin	62. p,p'-DDE	63. Cypermethrin-1
64. Malaoxon	65. Myclobutanil	66. Cyfluthrin-4
67. Alachlor	68. Oxyfluorfen	69. Etofenprox
70. Fenvalerate-1 / Fenvalerate-2 (Esfenvalerate)	71. Deltamethrin-1 (Tralomethrin deg.-1)
Qualitative and Quantitative Analysis
Qualitative assessment of the presence of pesticides in each sample, quantitative analysis was carried out to calculate the quantity of detected pesticides. For the purpose, the following formula was used:

Residue concentration (µg/g) = Csample (µg/L) x d /sample weight

Csample = instrument concentration of sample extract (µg/L). d = Dilution factor

Analysis by injecting calibrating standards of different concentration levels with samples in the same sequence was processed and concentrations of samples were calculated.

Health Risk Assessment

The assessment of risk associated with the consumption of each pesticide detected in the sample was monitored and evaluated. The health risk index was calculated by EDI and ADI. ADIvalues was used from literature studies. The EDI value was calculated by the average consumption of a person per day (kg/day) multiplying with residual concentration of pesticide (mg/kg) and dividing by the average weight of person (60 kg). The average consumption of vegetables for adults was 0.345 kg/person/day. The health risk index for each pesticide was calculated using the following formula (Hossain et al., 2015):

Health Risk Index = Estimated Daily Intake/Acceptable Daily Intake

EDI= __ concentration of detected pesticide residue X food consumption

body weight of an adult

Method Verification

QuEChERS method for extracting multi-residue pesticides in vegetable samples which was validated for recovery, repeatability and detection limits. The recovery of pesticides were from 70.5–119.6%, and the precision values (expressed as RSD, %) were below 20% for all of the pesticides (DG SANTE/11813/2017). The limit of detection (LOD) and limit of quantitation (LOQ) were calculated by multiplying the standard deviation of repeatability by factors of 3 and 6, respectively (Hovind et al., 2012). In this study, 161 pesticides from different chemical classes are commonly used in vegetables of four districts of Haryana. A total of 90 pesticides were analyzed by LC-MS/MS, and the remainders 71 of were analyzed by GC-MS/MS.

Evaluation by Commodity

The concentrations of pesticide residues in 102 vegetable samples, 46 soil and 50 water samples were analysed. A total of 161 different pesticides were screened for the presence in liquid chromatography–tandem mass spectrometry (LC-MS/MS) (90 pesticides) and gas chromatography–tandem mass spectrometry (GC-MS/MS) (71). Detectable residues were found in 109 samples (55.05%), while 89 samples (54.05%) were found to be residue-free. In total 30.39% vegetables, 95.65% soil and 68% water samples were found with detectable residues (Fig. 2).

Cypermethrin, Pendimethlin, Buprofezin, Oxyflurofen, Tubeconazole and Mycobutanil were detected residues with higher concentration. Alphacypermethrin, Hexaconazole and Carbendazim were detected residues with lower concentration. The detailed contaminations are given in Table.1.

Table 1

Chemical contamination level in analysed samples of vegetable, soil & water during 2019-20.
S.No..	Chemical Contamination(2019-20)			Contamination level
	Name of Chemical	Contamination level found (ppm)	Reference Range (ppm)	Low	Medium	High
1.	Chloropyrifos	0.002–2.023	0.2		✓
2.	Cypermethrin	0.001–24.546	0.2			✓
3.	Pendimethal in	0.005–1.091	0.05			✓
4.	Buprofenzin	0.016-1.0	0.01			✓
5.	Butachlor	0.008–0.27	0.05		✓
6.	Deltamethrin	0.010–0.708	0.05		✓
7.	Metalaxyl	0.008-0.70	0.01		✓
8.	Alpha Cypermethrin	0.0019–0.119	0.5	✓
9.	Hexaconazole	0.009–0.255	0.5	✓
10.	Propiconazole	0.008–0.287	-		✓
11.	Oxyflurofen	0.0016–5.727	-			✓
12.	Carbendizim	0.006–0.272	0.5	✓
13.	Tebuconazole	13.368	-			✓
14.	Mycobutanil	13.62	-			✓

From 25 samples of vegetables, 13 (52%) from Karnal district were found contaminated (Table. 2). Triazophos reported in higher concentration in cucumber (0.480 ppm) followed by cypermethrin with residual amount of 0.036 ppm. Heaxconazole (1,040 ppm), chlorpyrifos (0.035–0.039 ppm) and etofenprox(0.030 ppm) were reported in okra samples. In Rohtak 10 (41.6%) samples were found contaminated out of 24 samples collected. Two samples of cucumber were contaminated with metalaxyle (0.030–0.032 ppm) and carbendazim (0.035 ppm), Tomato and lobia samples were reported with cypermethrin (0.024 ppm), chlorfenpyr (0.104 ppm) and triazophos (0.387 ppm) contamination. Triazophos (0.010–0.012 ppm), propagrite (0.014 ppm), dimethoate (0.11 ppm)were reported in bottle gourd samples. Tori samples were reported with maximum contamination of thiamethoxam (0.155 ppm), dimethoate (0.142 ppm), linuron (0.020 ppm), malathion (0.047 ppm), pendimethalin (0.016 ppm), propagrite (0.649 ppm) and cyfluthrin (0.011 ppm). In Hisar district 38% of the samples were reported contaminated. Cucumber was reported to be maximum contaminated (7 samples) with metaxyl (0.010-0.053ppm). chlorfenpyr (0.038–0.043 ppm) and cyfluthrin(0.021–0.061 ppm) followed by cauliflower with cypermethrin (0.013 ppm) and carbendazim (0.135ppm), One tomato sample was reported to contain mycobutanil (0.010 ppm) pesticide residue.

Table 2

Pesticide contamination in vegetable samples collected from districts in Haryana
District	Commodity	Pesticide Detected	Conc. mg/Kg	ADI (mg/Kg body weight)	Samples above MRL	^*MRL
Rohtak (Total no. of samples = 24; Total contaminated samples = 10)	Tori	Thiomethoxom Dimethoate Linuron Malathion Pendimethalin Propargite Cypermethrin	0.155 0.142 0.020 0.047 0.016 0.649 0.015	- 0.002 - 0.03 - 0.01 0.05	- - - - - - -	- 2 - 3
	Bottle guard	Trizophos Propargite Dimethoate Trizophos	0.010 0.014 0.011 0.012	0.001 0.01 0.002 0.001	- - - -	- - 2 -
	Brinjal	BDL			-	-
	Chilli	Malaoxan	0.014	0.03	-	3
	Tomato	Chlorfenpyr Trizophos	0.104 0.387	0.2 0.001	- -	- -
	Lady Finger	BDL	-	-	-	-
	Cucumber	BDL	-	-	-	-
	Lobia	Cypermethrin	0.024	0.05	-	-
Hisar (Total no. of samples = 26; Total contaminated samples = 07)	Tomato	Myclobutanil	0.010	0.03	-	-
	Cucumber	Metalaxyl	(0.010–0.053)	0.08	-	-
	Cucumber	Cyfluthrin Chlofenopyr	(0.021–0.061) (0.038–0.043 )	0.02 0.05		- -
	Cauliflower	Cypermethrin Carbendazim	(0.013) 0.135	0.05 0.03	- -	- 0.5
	Cabbage	BDL	-	-	-	-
	Brinjal	BDL	-	-	-	-
Panipat (Total no. of samples = 27; Total contaminated samples = 8)	Cauliflower	Cypermethrin	0.017–0.025	0.05	-	-
	Brinjal	BDL	-	-	-	-
	Raddish	Chlorpyriphos Cypermethrin	2.023 0.062	0.01 0.05	-	- -
	Spinach	BDL	-	-	-
	Bottle Guard	BDL	-	-	-
	Ridge Guard	BDL	-	-	-
Karnal (Total no. of samples = 25; Total contaminated samples = 1)	Cauliflower	BDL	-	-	-
	Bitter Guard	BDL	-	-	-
	Cucumber	Trizophos Alpha- Cypermethrin	0.480 0.036	0.001 0.05
	Brinjal	BDL	-	-	-
	Bottle Guar	BDL	-	-	-
	Capsicum	BDL	-	-	-
	Sponge Guard	BDL	-	-	-
	Okra	Hexaconzole Chlorpyrifos	0.040 0.039	0.005 0.01	- -	- 0.2
^*FSSAI, Compendium_Contaminants_Regulations_28_01_2022.pdf

MRLs and ADI

Pesticide residue concentrations above the MRLs by EU regulations (National Sample Survey Office (NSSO) 2014) were identified in vegetables samples detected in a total of 31 samples (30.39%). ADI values were surpassed most often in tori, bottle guard, tomato, cucumber, raddish and okra samples. Calculated concentration of some of the pesticides like Dimethoate, Malathion, Propergite, Cypermethrin of tori samples, Trizophos, Propergite, Dimethoate of bottleguard, Trizophos of tomato and Cypermethrin detected in lobhia samples of district Rohtak exceeds the ADI limit. Cyfluthrin of cucumber, Carbendazim of cauliflower detected in district of Hisar, Chlorpyrifos and cypermethrin of raddish were detected in Panipat and Trizophos and Alpha-cypermethrin of cucumber and Hexaconazole and Chlorpyrifos of okra of Karnal district of Haryana. Non of the detected residue exceeds above MRL.

Soil

From 12 soils samples of Karnal district 8 (66.6%) samples were found contaminated with chlorpyrifos (0.018–0.058 ppm), Butachlor (0.014–0.124 ppm), Lambda-cyhalothrin (0.016ppm), Pendimethlin (0.013 ppm), Cypermethrin (0.003–0.826 ppm), Metaxyle (0.012 ppm), Propiconazole (0.008ppm) and Oxyflurofen 0.011 ppm ). In Panipat district 11 soil samples were collected and tested out of which 9 sample were found contaminated with Buprofezin (0.56 ppm), Cypermethrin (0.015–0.045 ppm), Pendimethalin (0,010-0.037 ppm), Hexaconazole (0.014 ppm), Fipronil sulphone (0.024 ppm), Oxyflorfen (0.091ppm), Emamectin benzoate (0.033 ppm), Lambda-cyhalothrin (0.013ppm), Metaxyle (0.032–0.305 ppm), Chlorpyrifos (0.013–0.014 ppm), Deltamethrin (0.010 ppm) and Propiconazole (0.010 ppm).

13 soil samples of Rohtak district were tested and all were detected with pesticide residue. 13 sample were found contaminated with Pendimethlin (0.0014–0.027 ppm), Hexaconazole 0.009 ppm), Fipronil sulphone (0.024 ppm), Metalaxyl (0.008 ppm), Chlorpyrifos (0.002–0.014 ppm), Deltamethrin (0.0017 ppm), Carbendazim (0.028–0.166 ppm), Difenconazole (0.009 ppm), Butachlor (0.008 ppm), Alpha-cpermethrin (0.019–0.119 ppm), Pretilachlor (0.009 ppm), Temephos (0.136ppm), Lambda cyhalothrin (0.0044–0.0283 ppm), Atrazine (0.0038 ppm), Oxyflurofen (0.0016 -0.0140ppm), Ethion (0.0017 ppm) and Thiophanate methyl (0.009-0.024ppm). All the 12 samples collected from Hisar districts were found contaminated with Cypermethrin (0.016–0.165 ppm), Carbendazim (0.006–0.272 ppm), Pendimethlin (0.010–0.017 ppm), Imidacloprid (0.017-0.022ppm), Thiophanate methyl (0.308 ppm), Hexaconazole (0.013 ppm), Metalaxyl (0.049 ppm), Chlorpyrifos (0.016–0.029 ppm), Pendimethlin (0.010–0.017 ppm), Ethion (0.008ppm), Tubeconazole (0.008ppm) and Mycobutanil (0.034 ppm).

Water

13 water samples were taken from Karnal district in which 2 (15.38%) samples were found residue of Cypermethrin (0.035pm) and Buprofezin (1.016 ppm). In Panipat district 18 water samples were collected and tested out of which 8 samples were detected with Cypermethrin (0.21 ppm), Pendimethlin (0.005 ppm),Chlorpyrifos (0.007-0,013 ppm), Deltamethrin (0.011–0.162 ppm), Alpha cypermethrin(0.013–0.042 ppm and Metribuzin (0.010 ppm). 13 water samples of Rohtak district were tested and in which 12 samples were detected with Cypermethrin (0.001ppm), Phorate (0.006 ppm), Metribuzin (0.34–3.9 ppm), Chlorpyrifos(0.2–0.55 ppm), Pendimethlin (0.26–0.29 ppm), Fipronil (0.91 ppm), Butachlor (0.18–0.27 ppm). Profenfos (0.29 ppm), Oxyfluorfen (0.34- 1.68ppm), Buprofezin ( 0.13-1.0 ppm), Trizofos (0.34–0.55 ppm), Propiconazole (0.31 ppm), Beta HCH (0.35ppm),Chlorthanonil (0.39 ppm), Mycobutanil (0.25 ppm),Lambda Cyhalothrin (0.04ppm), Permethrin (0.12 ppm) and Malathion (0.25 ppm). All the 12 samples collected from Hisar district were found contaminated with Chlorpyrifos (0.673–1.324 ppm), Butachlor (0.115–0.166 ppm), Oxyflrofen( 2.264–5.727 ppm), Cypermethrin (0.856-24.546ppm), Pendimethlin (0.124- 1.09ppm), Cyhalothrin (0.370 ppm), Propiconazole (0.135–0.287 ppm), Permethrin (0.284 ppm), Tebuconazole (13.368 ppm), Azoxystrobin (0.263 ppm), Iprobenfos (0.155ppm), Metalaxyl (0.700 ppm), Hexaconazole (0.255 ppm) and Mycobutanil (16.7^,22 ppm). The contamination of samples are represented in figure below

Pesticide Residue Evaluation

In this study, the concentrations of 161 different pesticides were determined in 14 different vegetable commodities, soil and water. From the total 198 samples (Vegetables, soil and water) i.e 109 samples were detected with pesticide residue. Of all detected pesticides Chlorpyrifos, Cypermethrin, Pendimethlin and Lambda cyhalothrin were found the most often. Triazophos reported maximum in Cucumber followed by Alpha Cypermetrin and Hexaconazole. Chlorpyrifos & Etofenprox were reported in Okra in Karnal. Chlorpyrifos was reported in very high concentration in Raddish followed Thiamethoxam and Azoxystrobin in Okra samples of Panipat. Cucumber samples were contaminated with Metalaxyle and Carbendazim. Tori samples of Rohtak were reported with maximum pesticide residue contamination of Thiamethoxam. Dimethoate. Linuron & Melathion. Cucumber samples collected from Hisar were reported maximumwith Metaxyle, Chlorfenpyr & Cyfluthrin followed-by Cauliflower with Cypermethrin and Carbendazim. Pesticide residue contamination level of soil of Karnal (52%) > Rohtak (41.6%) > Hisar (38%) > Panipat (29.62%). Also Chlorpyriphos, Buprofezin, Pretilachlor, Cypermethrin, Pendimethlin Deltamethrin, Lambda-cyhalothrin, Metalxyle were the major contaminates reported in soil samples of Karnal, Panipat, Rohtak & Hisar districts of Haryana. Pesticide contamination level of Rohtak (100%) ,Hisar(100%) > Panipat (81%) > Karnal (66%). Chlorpyriphos, Buprofezin, Pretilachlor, Cypermethrin, Pendimethlin, Deltamethrin. Lambda-cyhalothrin and Metribuzin were the major contaminates reported in water samples of Karnal, Panipat, Rohtak & Hisar. Pesticide residue contamination level in water samples of Hisar (100%) > Rohtak (92.3%) > Panipat (44.4%) > Karnal (15.38%). The contamination percent of tested samples (water, soil and vegetables) from Panipat, Hisar, Rohtak and Karnal are given in Fig. 4.

Pesticides detected in samples was confirmed by ion ratios and retention times from the matrix-matched standards with the reference values and most were within the required tolerances (± 30% and ± 0.1 min, respectively). According to the results obtained, fourteen pesticides were detected in different samples of vegetables, soil, and water. Detected pesticides were Chloropyrifos, Pendimethalin, Tebuconazole, Carbendizim, Oxyflurofen, Propiconazole, Hexaconazole, Alpha Cypermethrin, Metalaxyl, Deltamethrin, Butachlor, Buprofenzin, Metribuzin and Mycobutanil. Chlorpyrifos was present in Okra, raddish which protect crop from foliage insects and boring insects. Alpha cypermetnrin and Cyfluthrin present in cucumber is used to control pests. Tebuconazole, Propiconazole and Hexaconazole are pesticides used to protect the crop fungus. Linuron, Butachlor, Buprofenzin, Metribuzin are herbicide class of pesticides used to protects crops from surrounding herbs. Thiamethoxam works against sucking and chewing pests. Dimethoate detected in tori used to kill insects and pests by distrupting central and peripheral nervous system. Chlorphenapyr, Propergite residue present in tori, tomato, cucumber and brinjal used to protect crop from termites and other insects by inhibiting magnesium stimulated ATPase. Trizofos is an araricides and nematicides class whose residue present in bottle guard, tomato and cucumber. Among all these, the irrigation water from Hisar was more contaminated from other districts. Water samples contaminates with phorate, profenophos, Beta-HCH, Chlorothalonil, Permethrin and Iprobenfos were present but it was not present in vegetable and soil samples. Almost all the vegetables detected with pesticides residue where found below Maximum Residue Limit. However, the MRLs for the pesticide detected in soil samples are not reported. The exposure of pesticides causes irritation, headache, dizziness, nausea, allergies, asthma, nasal discharge, bronchitis, sinusitis, sneezing and also carcinogenic in the long run (Chensheng et al., 2006). The pesticide residue present in most of the water samples of Rohtak and Hisar but Karnal with the least. The Drinking Water Directive 98/83/EC²¹ sets a maximum concentration of 0.1 mg/L for each individual pesticide and their degradation products and 0.5 mg/L for the sum of all pesticides present in a sample. Almost all water samples from Rohtak and Karnal were above maximum concentrations sets by Drinking water Directive 98/83/EC. Extreme higher level of contamination of tubeconazole (13.37 mg/L), myclobutanil (16.322 mg/L) and Oxyflorfen (2.32–5.72 mg/L) were present in water samples of Hisar Districts. Only one water samples was contaminated with buprofezin (1.02 mg/L) at higher concentration level.

Risk Assessment

Estimation of risk assessment can be calculated by the detected pesticide residues in samples, whereas an exposure analysis was carried out by monitoring the results. The EDI for vegetables was 0.45. Hazard index for Trizophos in tomato of Rohtak district and cucumber of Panipat district was 2.225, and 2.760 respectively. Hazard index for Chlorpyrifos in raddish of panipat district was 1.163. Hence HRI > 1 indicates the potential health risk. Pesticides like Dimethoate, Malathion, Propergite, Cypermethrin of tori samples, Propergite, Dimethoate of bottleguard, Cypermethrin detected in lobia samples of district Rohtak, Cyfluthrin of cucumber, Carbendazim of cauliflower detected in district of Hisar, cypermethrin of raddish were detected in Panipat and Alpha-cypermethrin of cucumber and Hexaconazole of okra of Karnal district of Haryana had no risk due to the values below 1. The results indicate that some samples pesticidal residue concentration was higher but had no health risks (Table 3).

Table 3

Health risk assessment of pesticide residues in vegetables.
District	Commodity	Pesticide Detected	Conc. mg/Kg	MRL	ADI	EDI	HRI	If HRI > = 1 then ,health risk Yes
Rohtak		Thiomethoxom	0.155		-			No
		Dimethoate	0.142	2	0.002	0.001	0.408	No
	Tori	Linuron	0.02		-			No
		Malathion	0.047	3	0.03	0.000	0.009	No
		Pendimethalin	0.016		-			No
		Propargite	0.649		0.01	0.004	0.373	No
		Cypermethrin	0.015		0.05			No
	Bottle guard	Trizophos	0.01		0.001	0.000	0.058	No
		Propargite	0.014		0.01	0.000	0.008	No
		Dimethoate	0.011	2	0.002	0.000	0.032	No
		Trizophos	0.012		0.001	0.000	0.069	No
	Brinjal	BDL						No
	Chilli	Malaoxan	0.014	3	--	0.000		No
	Tomato	Chlorfenpyr	0.104		0.2	0.001	0.003	No
		Trizophos	0.387		0.001	0.002	2.225	Yes
	Lady Finger	BDL	-		-			No
	Cucumber	BDL	-		-			No
	Lobia	Cypermethrin	0.024		0.05	0.000	0.003	No
Hisar	Tomato	Myclobutanil	0.01		0.03	0.000	0.002	No
	Cucumber	Metalaxyl	(0.010–0.053)		0.08	0.000	0.004	No
								No
		Cyfluthrin	(0.021–0.061)		0.02	0.000	0.018	No
								No
						0.000		No
		Chlofenopyr	(0.038–0.043 )		0.05	0.000	0.005	No
						0.000		No
						0.000		No
	Cauliflower	Cypermethrin	-0.013		0.05	0.000	-0.001	No
		Carbendazim	0.135	0.5	0.03	0.001	0.026	No
	Cabbage	BDL	-		-			No
	Brinjal	BDL	-		-			No
Panipat	Cauliflower	Cypermethrin	0.017–0.025		0.05	0.000	0.003	No
						0.000		No
	Brinjal	BDL	-		-			No
	Raddish	Chlorpyriphos	2.023	0.2	0.01	0.012	1.163	Yes
		Cypermethrin	0.062		0.05	0.000	0.007	No
	Spinach	BDL	-		-			No
	Bottle Guard	BDL	-		-			No
	Ridge Guard	BDL	-		-			No
Karnal	Cauliflower	BDL	-		-			No
	Bitter Guard	BDL	-		-			No
	Cucumber	Trizophos	0.48		0.001	0.003	2.760	No
		Alpha- Cypermethrin	0.036		0.05	0.000	0.004	No
	Brinjal	BDL	-		-
	Bottle Guar	BDL	-		-
	Capsicum	BDL	-		-
	Sponge Guard	BDL	-		-
	Okra	Hexaconzole	0.04		0.005	0.000	0.046	No
		Chlorpyrifos	0.039	0.2	0.01	0.000	0.022	No

Out of 198 samples (Soil + Water + Vegetables) 109 (55.05%) samples were found above maximum residue limit (MRL), LOQ (Limit of quantification) for vegetable and soil samples are 0.01 ppm and for water it is 0.1ppb. Cypermethrin, Chlorpyriphos, triazophos, buprofezin, dimethoate, cyhalothrin, emamectin benzoate, thiomethoxam are major contaminatents from the insecticides category. Major herbicide reported is pendimethlin, oxyflrofen„ atrazine, pretilachlor. Major fungicides reported in soil, water and vegetables are Hexaconazole ,Propiconazole, Mycobutanil, Carbendazim, Difenconazole and Thiophanate methyl in vegetables. Karnal district samples shows maximum contaminated samples of soil & water. Hisar and Rohtak disrtrict samples of soil and water were reported with pesticide residue and level of contamination in Hisar water samples were of serious concern as Cypermethrin was 24.5 ppm Tebuconazoie 13.368 ppm and Mycobutanil at 13.62 ppm level. The study revails that pesticides such as Trizophos and Chlorpyrifos had health risk indices above HRI greater than 1. The results indicates health risk indices of Trizophos and Chlorpyrifos ranging from 1.16 to 2.760 in vegetables which causes serious human health risk.

Acknowledgements

The authors sincerely thanks to Director IPFT and Directorate of Horticulture Haryana, for providing necessary facilities and funds respectively to carry out this work.

Author contribution

Sudeep Mishra: Extraction of samples, sample preparation, analysis of samples and drafting of article.

Mukesh Kumar Singh: Extraction of samples, sample preparation and analysis of samples.

Mahesh Kumar Saini: Extraction of samples, sample preparation and analysis of samples.

Samsul Alam: Conception and designs of works, statistical analysis and presentation of the data.

Lalitesh Kumar Thakur: Conception and designs of works, statistical analysis, presentation of the data and critically revising the article.

All authors read and approved the final manuscript.

Data availability The data releated to the study are made available from the corresponding author on reasonable request.

Funding: Directorate of Horticulture Haryana has provided samples and funds for their analysis.

Ethical approval No any animals were harmed nor be a part of this study.

Conflict of interest The authors declare no conflict of interests.

Competing interests The authors declare that they have no competing interests.

https://www.icfa.org.in/assets/doc/reports/haryana-agriculture-and-farmers.pdf
Abhilash P.C. and Singh N. (2009). A study on Pesticide use and application: an Indian scenario.
Anastassiades, M., Lehotay, S. J., Stajnbaher, D., Schenck F. J. (2003). Fast and easy multiresidue method employing acetonitrile extraction/partitioning and “dispersive solid-phase extraction” for the determination of pesticide residues in produce. Journal of AOAC International, 86, 412-431.
Bhardwaj, Tulsi and Sharma, J.P. (2013). Impact of pesticidesapplication in agricultural industry: An Indian scenario. International Journal of Agriculture and Food Science Technology, l4(8), 817-822.
Chensheng, L., Toepel, K., Irish, R., Fenske, R. A., Barr, D. B., and Bravo, R. (2006). Organic diets significantly lower children’s dietry exposure to organophosphorus pesticides. Environmental Health Perspectives, 114(2), 260–263.
Choi, S. M., Yoo, S. D., Lee, B. M. (2004). Toxicological characteristics of endocrine disrupting chemicals: Developmental toxicity, carcinogenicity, and mutagenicity. Journal of Toxicology and Environmental Health, Part B, 7, 1–24.
Directorate General Health and Food Safety (DG SANTE). Guidance Document on Analytical Quality Control and Method Validation Procedures for Pesticide Residues and Analysis in Food and Feed. SANTE/11813/2017; Directorate General Health and Food Safety: Luxembourg, 2017.
FAO/WHO (Food and Agriculture Organization/World Health Organization). Food Standards Programme. In Proccedings of Codex Alimentarius Commission. Twenty-Seventh Session, Geneva, Switzerland, 28 June–03 July 2004, pp. 1–103.
Galloway, T., Handy, R. (2003). Immunotoxicity of organophosphorous pesticides. Ecotoxicology, 12, 345–363.
Hayat, K. Ashfaq, M. Ashfaq, U. Saleem, M. A. (2011). Determination of pesticide residues in blood samples of villagers involved in pesticide application at District Vehari (Punjab), Pakistan. African Journal of Environmental Science and Technology, 4, 666–684.
Hovind, H., Magnusson, B., Krysell, M., Lund, U., Mäkinen, I. Internal quality control—Handbook for chemical laboratories. NT TR 2012, 569, 04038.
Hossain M. S., Fakhruddin, A. N. M., Chowdhury, A. Z. M., Rahman, M. A. and Khorshed, A. M. (2015). Health risk assessment of selected pesticide residues in locally produced vegetables of Bangladesh. International Food Research Journal, 22(1), 110–115.
Lehotay, S .J. (2007). Determination of pesticide residues in foods by acetonitrile extraction and partitioning with magnesium sulphate: Collaborative study. Journal of AOAC International, 90(2): 485-520.
Mishra, S., Singh, M. K., Saini, M. K., Vikash, Alam, S. et al., (2019). Pesticide residues in vegetable, soil and water samples collected from some districts of Haryana. Agriculture Research Journal, 56 (2), 261-266.
Mishra, S., Deotale, V. D., Pande, M. A., et al., (2016). GC-MS analysis of multiclass pesticide residues found in farmland water of Haryana districts. World Journal of Pharmaceutical Research, 5 (8): 744-752.
National Sample Survey Office (NSSO) 2014. Household consumption of various goods and services, India. Retrieved in April, 2020 from http://www.icssrdataservice.in/datarepository/index.php/ctalog/MoSPI-NSS.
Pesticide Properties Data Base A to Z List of Pesticide Active Ingredients. Available online: https://sitem.herts.ac.uk/aeru/ppdb/en/atoz.htm (accessed on 15 July 2018).
Sushil, S. N. (2016). Emerging Issues of Plant Protection in India. Natural Resource Management: Ecological Perspectives. International Conference, SKUAST, Jammu.
Wood, A. M. Compendium of Pesticide Common Names According to ISO 1750. Available online: http://www.alanwood.net/pesticides/ (accessed on 15 June 2018).

No competing interests reported.

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Evaluation and assessment of risk of pesticide residue in vegetables, soil and water samples of few district of Northern India

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