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9 protocols using ethion

1

Pesticide Standards and Calibration Curves

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Malathion, parathion, sulfotep, fenthion, disulfoton, demeton-O, chloropyrifos and bifenthrin certified analytical standards were purchased from AccuStandard (New Haven, CT, USA). In addition, ethion, carbofuran and azoxystrobin standards were acquired from Sigma-Aldrich (St. Louis, USA) and permethrin acquired from Supelco (St. Louis, USA). All standards presented purity grade of 97% or higher. Acetonitrile (99.99%) (JT Baker, USA), dichloromethane (99.9%) (JT Baker, USA) and methanol (99.9%) (Merck, Darmstadt, Germany) used in this study were spectroscopic and chromatographic grades.
Stock solutions were prepared in methanol at 500 mg L−1 for carbofuran, diazinon, chloropyrifos, ethion, bifenthrin, permethrin and azoxystrobin. The other stock solutions were at 960 mg L−1 (demeton-O), 984 mg L−1 (fenthion), 985 mg L−1 (sulfotep), 990 mg L−1 (molinate), 998 mg L−1 (parathion) and 1000 mg L−1 for Malathion. From the stock solutions, a mix working solution (1000 µg L−1) containing the 13 pesticides was prepared by dilution in methanol. On the other hand, matrix-matching calibration curves composed of seven concentration levels (0.5, 1.5, 2.5, 3.5, 4.5, 5.5, 6.5, and 7.5 µg L−1) were prepared for each compound adding variable volumes of mix working solutions in extracts of PM2.5 blank filters.
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2

Analytical Determination of Insecticides

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Standards for the carbamate insecticides methomyl, carbaryl, and methiocarb, and the organophosphate insecticides fenthion, chlorpyrifos, and ethion were purchased from Sigma-Aldrich (St. Louis, MO, U.S.A.). HPLC-grade methanol and acetone were purchased from Merck (Darmstadt, Germany), while hexane and ethyl acetate (EA) were purchased from J.T. Baker (Phillipsburg, NJ, U.S.A.). Acetic acid was purchased from Sigma-Aldrich (St. Louis, MO, U.S.A.). Distilled deionized water (purified with a PURE-LAB Classic UV from ELGA, Marlow, U.K.) was used to prepare the electrospray solution. All chemical reagents and solvents were used without additional purification. Human serum was purchased from the Innovative Research Incorporation (Oakland, CA, U.S.A.) and diluted 1000 times in methanol for analysis.
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3

Pesticide Extraction and Quantification

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Parathion-methyl, chlorpyrifos, captan, procymidone, α-endosulfan, prothiofos, cyproconazole, ethion, triazophos, and phosmet of 98% w/w purity grade were purchased from Sigma-Aldrich (St. Louis, MO, USA). The choice of pesticides was based on their use on the region of samples collection. A work solution of 20.00 mg L−1 was prepared by the appropriate dilution in HPLC-grade methanol, Sigma-Aldrich (St. Louis, Missouri, United States). This work solution was used for the matrix spike in different concentration levels (5.00 to 160.00 μg L−1) to optimize the extraction conditions during the validation study. Calibration standards were prepared at 5.00, 10.00, 20.00, 40.00, 80.00, and 160.00 μg L−1 concentrations using ultrapure water produced in a Purelab UVMK2 purifier from Elga (Marlow, Buckinghamshire‎‎, England). 1-Octanol HPLC grade was purchased from Sigma-Aldrich (St. Louis, Missouri, United States), ethyl decanoate, acetonitrile, and ethyl octanoate were purchased from J. T. Baker (Xalostoc, Edo MEX, Mexico). Hollow fiber was purchased from Underlying Performance Co. (Wuppertal, Germany).
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4

Comprehensive Analysis of Pesticide Residues

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CRM for organochlorine pesticides comprised a mixture of 20 compounds viz. α-BHC, β-BHC, γ-BHC, Δ-BHC, heptachlor, Aldrin, heptachlor epoxide, trans-chlordane, cis-chlordane, endosulfan-I, endosulfan-II, endosulfan sulfate, 4,4′-DDE, 4,4′-DDD, 4,4′-DDT, dieldrin, endrin, endrin aldehyde, endrin ketone and methoxychlor; a herbicide mix of 5 compounds viz. atrazine, simazine, alachlor, metolachlor and butachlor and were procured from Sigma Aldrich (USA).
Individual standards for hexachlorobenzene, and several organophosphate pesticides including ethion, disulfoton, malaoxon, fenthion, phosalone, methyl parathion, methyl chlorpyrifos, ethyl parathion, fenitrothion, propazine and methyl paraoxon, were all procured from Sigma Aldrich (USA). Standards for malathion, acephate, phosphamidon, captafol, diazinon and monocrotophos were procured from Accustandard, Inc. (USA). A mixture of pyrethroid compounds including Cyfluthrin, L-cyhalothrin, cypermethrin-I, cypermethrin-II, cypermethrin-III, deltamethrin, pendimethalin, permethrin-I, permethrin-II, fenvalerate, and tefluthrin were also purchased from Accustandard, Inc. (USA).
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5

Liquid Crystal Biosensing Protocol

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All the chemicals used were of analytical grade and the aqueous solutions were prepared with deionized (DI) water (resistivity of 18.2 MΩ.cm). Anhydrous n-heptane, CH2Cl2, H2SO4 (95 %), H2O2 (30%), CH3OH, and C2H5OH were procured from Daejung Chemicals & Metals Co., Ltd. of South Korea. Nematic liquid crystal 4-cyano-4’-pentylbiphenyl (5CB) was purchased from Tokyo Chemical Industrial Co., Ltd. of Japan. Malathion, ethion, fenthion, fenobucarb, carbofuran, phosmet, CTAB, phosphate-buffered saline (PBS, pH 7.4), and trichloro(octyl)silane (OTS) were obtained from Sigma-Aldrich, USA. The DNA aptamer with the sequence 5’-ATCC GTCA CACC TGCT CTTA TACA CAAT TGTT TTTC TCTT AACT TCTT GACT GCTG GTGT TGGC TCCC GTAT-3’ was synthesized by Mbiotech (Hanam, Korea).
Polarized optical images of 5CB were captured using a digital camera (DS-2Mv, Nikon, Tokyo, Japan) affixed to a polarizing optical microscope (Eclipse LV100 POL, Nikon, Japan). All images were taken in transmission mode using a 4× objective lens. The gray-scale intensities (GIs) of the optical images were calculated using Adobe Photoshop CC2019 software (Adobe Inc., San Jose, CA, USA). Tests were conducted in at least triplicate for each measurement.
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6

Pesticide Standards for Analytical Methods

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Diazinon, methyl-parathion, fenitrotion, malathion, chlorpyrifos, triadimephon, penconazole, imazalil, myclobutanil, ethion, trifloxystrobin, propiconazole, bromopropylate, lindane, endosulfan, aldrin, heptachlor epoxide A, trans-chlordane, dieldrin, endrin, p,p′-DDT, and p,p′-DDD standards of high purity (>98%) were supplied by Sigma-Aldrich (Seelze, Germany). The stock solutions (1 g l -1 ) were prepared by dissolving the standards in methanol HPLC grade (99.9%) from Sintorgan (Buenos Aires, Argentina) and stored under freezing condition (-18 °C) in dark bottles sealed with PTFE/silicone caps. The working standard solutions (50 mg l -1 ) were prepared in hexane HPLC grade (99.9%) purchased by Sintorgan (Buenos Aires, Argentina). Deionized water obtained from an E-pure water purification system (Barnstead/Thermolyne, Bedford, MA, USA) was used for physical-chemical analysis.
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7

Trace-level Pesticide Extraction

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LC-grade methanol and acetonitrile were purchased from TEDIA. The analytical standards (ethoprophos, coumaphos, profenofos, diazinon, ethion, parathion, phosalone, quinalphos, dicrotophos, purity >95%) were purchased from Sigma Aldrich. Azinphos methyl, tribuphos (purity >95%) were from Dr. Ehrenstorfer Gmbh. Other details are given in Table 1. The analytical-reagent grade anhydrous magnesium sulfate (MgSO4), sodiumchloride (NaCl) was from Kermel. The powder of C18 and primary secondary amine (PSA, 50 μm) were purchased from Agela Technologies. The guarantee dreagent-grade formic acid was purchased from ROE. Ultrapure water was obtained from a Milli Q purification system (Millipore, Molsheim). Individual pesticide stock standard solutions (1000 mg L -1 ) were prepared in methanol and stored at -18°C. A multistandard mixture containing 10 mg L -1 of each pesticide was prepared in methanol and stored at 4°C for less than 3 months. Mixed multi-standard working solutions were prepared fresh to avoid degradation of pesticides. The concentrations of the multi-standard working solutions were 10, 25, 50, 100 and 250 μg kg -1 .
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8

Pesticide Residue Analysis Protocol

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All solvents like n-hexane, acetone and ethyl acetate (HPLC grade) were purchased from Sigma-Aldrich. Co. USA, Spectrochem Pvt. Ltd. India and were glass distilled before use. acetone was refluxed over potassium permanganate for 4 hr and then distilled. Sodium chloride (NaCl), anhydrous sodium sulphate (Na2SO4) and magnesium sulphate (MgSO4) were procured from Himedia Pvt. Ltd. India. Before use sodium sulphate (Na2SO4) and magnesium sulphate (MgSO4) were purified with acetone and baked for 4 hr @ 600°C in muffle Furness to remove possible phthalate impurities. Primary secondary amine (PSA) bondasil 40 µm part 12213024 of Varian was used.
Pesticide standards (Dichlorvos 98.9%, phorate 96.0%, phorate-sulfone 96.8%, phorate-sulfoxid 94.8%, Dimethoate 99.6%, diazinon, methyl-parathion 99.7%, chlorpyrifos-methyl 99.9%, fenitrothion 95.2%, Malathion 97.2%, chlorpyriphos 99.9%, chlorofenvinfos 98.7%, profenofos 95.0%, Ethion 97.8%, Edifenphos 99.9%, anilophos 97.5% and phosalone 95.2%) were purchased from Supelco Sigma Aldrich USA, Fluka Sigma-Aldrich Schweis and Rankem Pvt. Ltd. New Delhi, India.
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9

Pesticide Standards Characterization

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Pesticides standards (chlorfenvinphos 97.5%, chlorpyrifos 99.9%, diazinon 99.0%, fenthion 98.3%, malathion 97.2%, parathion 99.5%, phosalone 99.5%, pirimiphosmethyl 99.5% and quinalphos 96.2%) were purchased from Fluka. Dimethoate 96.2% and ethion 99.3% (the later was chosen as internal standard as its commercialization in Portugal is prohibited since 2003), were purchased from Supelco (Saint Louis, USA). Ammonium acetate, acetic acid, potassium chloride, sodium chloride, chloroform, diethyl ether, n-hexane, isooctane, potassium dihydrogen phosphate, di-sodium hydrogen phosphate, isopropanol were purchased from Merck Co. (Darmstadt, Germany). Acetonitrile and methanol were purchased from Fisher Chemical and Sigma-Aldrich, respectively. Water was purified by a Milli-Q system obtained from Millipore (Molsheim, France). All solvents were of analytical or gradient grade. Extraction procedures were performed in a manual SPE equipment, using Oasis ® HLB (3 cc, 60 mg) and Sep-Pak ® C18 (3 cc, 500 mg) SPE cartridges, obtained from Waters™ (WATERS Corporation, USA).
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