Multi pesticide residue analysis in tobacco by GCMS/MS using QuEChERS as an extraction method
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1 PO-CON463E Multi pesticide residue analysis in tobacco by GCMS/MS using QuEChERS as an extraction method ASMS 04 TP76 Durvesh Sawant (), Dheeraj Handique (), Ankush Bhone (), Prashant Hase (), Sanket Chiplunkar (), Ajit Datar (), Jitendra Kelkar (), Pratap Rasam (), Kaushik Banerjee (), Zareen Khan () () Shimadzu Analytical (India) Pvt. Ltd., A/B Rushabh Chambers, Makwana Road, Marol, Andheri (E), Mumbai-40009, Maharashtra, India. () National Referral Laboratory, National Research Centre for Grapes, P.O. Manjri Farm, Pune-4307, Maharashtra, India.
2 Introduction India is the world s second largest producer (after China) and consumer (after Brazil) of tobacco with nearly $ 00.4 million revenue generated annually from its export. [] In countries like India, with tropical-humid climate, the incidences of insect attacks and disease infestations are frequent and application of pesticides for their management is almost obligatory. Like any other crop, tobacco (Nicotiana tabacum Linn.), one of the world s leading high-value crops, is also prone to pest attacks, and the farmers do apply various pesticides as a control measure. The residues of pesticides applied on tobacco during its cultivation may remain in the leaves at harvest that may even sustain post harvest processing treatments and could appear in the final product. Thus, monitoring of pesticide residues in tobacco is an important issue of critical concern from public health and safety point of view demanding implementation of stringent regulatory policies. [] To protect the consumers by controlling pesticide residue levels in tobacco, the Guidance Residue Levels (GRL) of 8 pesticides have been issued by the Agro-Chemical Advisory Committee (ACAC) of the Cooperation Center for Scientific Research Relative to Tobacco (CORESTA). Tobacco is a complex matrix and hence requires selective extraction and extensive cleanup such as QuEChERS (Quick Easy Cheap Effective Rugged Safe) to ensure trace level detection with adequate precision and accuracy. The objective of the present study was to develop an effective, sensitive and economical multi-pesticide residue analysis method for 03 pesticides in tobacco as listed in Table. Figure. Dried tobacco Method of Analysis Extraction of pesticides from tobacco Extraction of pesticides was done using QuEChERS method, as described below. [3] Take g of dry powdered tobacco leaves (Figure ). Add 8 ml of water containing 0. % acetic acid. Homogenize the sample and Keep it for 30. Add 0 ml ethyl acetate. Immediately, put 0 g sodium sulfate. Homogenize it thoroughly at 000 rpm for. Centrifuge at 000 rpm for for phase separation. Draw 3 ml of ethyl acetate upper layer from the extract for further cleanup.
3 Add ml toluene to it and vortex for 0.. Add cleanup mixture [PSA (0 mg), C8 (0 mg), GCB (7 mg) and anhydrous MgSO 4 (300 mg)] and vortex for. Centrifuge the mixture at 7000 rpm for 7. Collect the supernatant and filter through a 0. µm PTFE membrane filter. Inject.0 µl of the clean extract into GCMS-TQ8030 (Figure ). Figure. GCMS-TQ8030 Triple quadrupole system by Shimadzu Key Features of GCMS-TQ8030 ASSP (Advanced Scanning Speed Protocol) enables high-speed scan and data acquisition for accurate quantitation at 0,000 u/sec Capable of perforg simultaneous Scan/MRM UFsweeper technology efficiently sweeps residual ions from the collision cell for fast, efficient ion transport ensuring no cross-talk Two overdrive lenses reduce random noise from helium, high-speed electrons and other factors to improve S/N ratio Flexible platform with EI (Electron Ionization), CI (Chemical Ionization), and NCI (Negative Chemical Ionization) techniques Full complement of acquisition modes including MRM, Scan/MRM, Precursor Ion, Product Ion and Neutral Loss Scan 3
4 4 Table. List of pesticides,6-dichlorobenzamide -Phenylphenol 3,4-Dichloraniline 3-Chloroaniline 4-Bromo -Chloro phenol 4,4-Dichlorobenzophenone Acetochlor Acrinathrin Alachlor Aldrin Azinphos-ethyl Azinphos-methyl Azoxystrobin Barban Beflubutamid Benfluralin Benoxacor Beta-endosulfan Bifenox Bifenthrin Bitertanol Boscalid Bromacil Bromophos-ethyl Bromopropylate Bromuconazole- Bromuconazole- Butralin Butylate Carbaryl Carbofuran Carfentrazone Chlordane-trans Chlordecone Chlorfenvinphos Chlormephos Chlorobenzilate Chloroneb Chlorothalonil Chlorpyriphos-ethyl Chlorpyriphos-methyl Chlorpyriphos-oxon Chlorthal-dimethyl Cinidon-ethyl Cis-,,3,6 tetrahydrophthalimide Clodinafop propargyl Clomazone Crimidine Cyanophos Cyfluthrin- Cyfluthrin- Cyfluthrin-3 Cyfluthrin-4 Cyhalofop-butyl Cypermethrin- Cypermethrin-3 Cypermethrin-4 Cyprodinil Delta-HCH Demeton-s-methyl Demeton-S-methyl sulphone Dialifos Diazinon Dichlobenil Dichlofluanid Diclofop Dicloran Dieldrin Diethofencarb Difenoconazole- Difenoconazole- Diflubenzuron Diflufenican Dimethipin Dimethomorph- Dimethomorph- Dimoxystrobin Diniconazole Dinoseb Dinoterb Dioxathion Edifenfos Endosulfan sulphate Endrin Epoxiconazole Ethalfluralin Ethoprophos Etoxazole Etridiazole Etrimfos Famoxadone Fenamidone Fenarimol Fenbuconazole Fenchlorphos Fenchlorphos oxon Fenhexamid Fenobucarb Fenoxycarb Fenthion sulphoxide Fenvalerate Fipronil Fipronil sulphone Flucythrinate- Flucythrinate- Flufenacet Flumoixazine Fluquinconazole Flurochloridone- Flurochloridone- Flutolanil Flutriafol Fluxapyoxad Folpet Fuberidazole Heptachlor Hexaconazole Iprobenfos Isoprocarb Isoprothiolane Isopyrazam Isoxaben Lactofen Lambda-cyhalothrin Malaoxon Malathion Mepanipyrim Mepronil Metalaxyl Metalaxyl M Metazachlor Metconazole Methabenzthiazuron Methacrifos Methidathion Methiocarb Metholachlor-s Methoxychlor Metribuzin Mevinphos Monolinuron Myclobutanyl Napropamide Nitrapyrin Oxadiargyl Oxadiazon Oxycarboxin p,p-dde Parathion-ethyl Parathion-methyl Penconazole Pencycuron (Deg.) Pendimethalin Permethrin- Permethrin- Pethoxamid Phosalone Phosmet Pirimicarb Pretilachlor Procymidone Profenofos Propanil Propaquizafop Propazine Propham Propiconazole- Propisoclor Propyzamide Proquinazid Pyraflufen-ethyl Pyrazophos Pyrimethanil Pyriprooxyfen Pyroquilon Quinoxyfen Simazine Spirodiclofen Sulfotep Swep Tebufenpyrad Tebupirimfos Tebuthiuron Tefluthrin Terbacil Tetraconazole Tetradifon Thiobencarb Tolylfluanid Tralkoxydim Triadimefon Tri-allate Triazophos Tricyclazole Trifloxystrobin Triflumizole Triflumuron Trifluralin Triflusulfuron Triticonazole Valifenalate Vinclozolin Zoxamide (Deg.)
5 GCMS/MS Analytical Conditions The analysis was carried out on Shimadzu GCMS-TQ8030 as per the conditions given below. Chromatographic parameters Column : Rxi-Sil MS (30 m L x 0. mm I.D.; 0. µm) Injection Mode : Splitless Sampling Time :.0 Split Ratio :.0 Carrier Gas : Helium Flow Control Mode : Linear Velocity Linear Velocity : 40. cm/sec Column Flow :. ml/ Injection Volume :.0 µl Injection Type : High Pressure Injection Total Program Time : 4.87 Column Temp. Program : Rate (ºC /) Temperature (ºC) Hold time () Mass Spectrometry parameters Ion Source Temp. : 3 ºC Interface Temp. : 8 ºC Ionization Mode : EI Acquisition Mode : MRM Results For MRM optimisation, well resolved pesticides were grouped together. Standard solution mixture of approximately ppm concentration was prepared and analyzed in Q3 scan mode to detere the precursor ion for individual pesticides. Selected precursor ions were allowed to pass through Q & enter Q, also called as Collision cell. In Collision cell, each precursor ion was bombarded with collision gas (Argon) at different energies (called as Collision Energy-CE) to produce fragments (product ions). These product ions were further scanned in Q3 to obtain their mass to charge ratio. For each precursor ion, product ion with highest intensity and its corresponding CE value was selected, thereby assigning a characteristic MRM transition to every pesticide. Based on MRM transitions, the mixture of 03 pesticides was analyzed in a single run (Figure 3). Method was partly validated for each pesticide with respect to linearity (0. to ppb), reproducibility, LOQ and recovery. The validation summary for two pesticides namely Mevinphos and Parathion-ethyl (Sr. Nos.40 and 49 in Table ) is shown in Figures 4 and. The summary data of linearity and LOQ for 03 pesticides is given in Table and 3 respectively.
6 (x00,000) Figure 3. MRM Chromatogram for 03 pesticides mixture Calibration overlay Linearity curve Recovery overlay (x0,000). Area(x00,000) (x0,000) MRM : 9.00>7.00 MRM : 9.00> Post extraction spike Pre extraction spike Conc Linearity (R ) LOD (ppb) LOQ (ppb) S/N at LOQ % RSD at LOQ (n=6) % Recovery at LOQ Figure 4. Summary data for mevinphos Calibration overlay Linearity curve Recovery overlay (x0,000) Area(x00,000) 8.0 (x,000) 3. MRM : 9.0> MRM : 9.0> Post extraction spike Pre extraction spike Conc Linearity (R ) LOD (ppb) LOQ (ppb) S/N at LOQ % RSD at LOQ (n=6) % Recovery at LOQ Figure. Summary data for parathion-ethyl 6
7 Table. Linearity Summary Table 3. LOQ Summary Linearity (R ) Number of pesticides LOQ (ppb) Number of pesticides % RSD range (n=6) S/N Ratio range % Recovery range Conclusion A highly sensitive method was developed for quantitation of 03 pesticides in complex tobacco matrix by using Shimadzu GCMS-TQ8030. The MRM method developed for 03 pesticides can be used for screening of pesticides in various food commodities. For 90 % of the pesticides, the LOQ of 0 ppb or below was achieved. Ultra Fast scanning, UFsweeper and ASSP features enabled sensitive, selective, fast, reproducible, linear and accurate method of analysis. Reference [] Tobacco Board (Ministry of Commerce and Industry, Government of India), Exports performance during 03-4, (04),. [] CORESTA GUIDE Nº, The concept and implementation of cpa guidance residue levels, (03), 4. [3] Zareen S Khan, Kaushik Banerjee, Rushali Girame, Sagar C Utture et al., Journal of Chromatography A, Volume 343, (04), 3. First Edition: June, 04 For Research Use Only. Not for use in diagnostic procedures. The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. The information contained herein is provided to you "as is" without warranty of any kind including without limitation warranties as to its accuracy or completeness. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication. This publication is based upon the information available to Shimadzu on or before the date of publication, and subject to change without notice. Shimadzu Corporation, 04
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Comprehensive Quantitation and Identification of Pesticides in Food Samples Using the SCIEX UltraLC 100 and the SCIEX QTRAP 4500 System André Schreiber SCIEX, Concord, Ontario, Canada Overview Liquid Chromatography
Index 1 Residual Pesticides 3 Food Additives 4 Others 2 Veterinary Drugs (Antibiotics and Synthetic Antibacterials) 5 Reference Materials
C9-E7A Index Residual Pesticides. Analysis of 4 Pesticides With Capillary GC-FTD and GC-ECD () - GC Analysis of 4 Pesticides With Capillary GC-FTD and GC-ECD () - GC Analysis of 4 Pesticides With Capillary
Determination of pesticides in baby food by UHPLC/MS/MS using the Agilent 1290 Infinity LC system and the Agilent 6460 triple quadrupole LC/MS
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תוספת שלישית )תקנה 1, הגדרת "שארית"(
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DETERMINATION OF PESTICIDES IN FOOD USING UPLC WITH POLARITY SWITCHING TANDEM QUADRUPOLE LC/MS/MS
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GC and GC-MS Applications for Food Safety Analysis
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Expect more productivity
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Screening for Hundreds of Pesticide Residues Using a GC/Q-TOF with an Exact Mass Pesticide Database in Food
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Application Note GCMS-01 Comparison of Ionization Techniques for the Analysis of Trace-Level Pyrethroid Insecticides by GC-MS/MS
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Advanced Data Acquisition and Data Processing Workflows to Identify, Quantify and Confirm Pesticide Residues
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Reduce Cost of Pesticide Residue Analysis
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Date ACCURATE-MASS DATABASE FOR SCREENING OF PESTICIDE RESIDUES IN FRUITS AND VEGETABLES BY GAS CHROMATOGRAPHY TIME-OF-FLIGHT MASS SPECTROMETRY
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Quantitative Determination of Dioxins in Drinking Water by Isotope Dilution using Triple Quadrupole GC-MS/MS
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Validation Report 18
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Fast Screening and Quantitation of Perfluorinated Sources from Textiles using Chemical Ionization GC-MS
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Analytical Test Report 10
OLCC #022892C 9 SE Division St. Laboratory ID: 805024-02 Batch ID: AC Date Sampled: 05/07/8 Date Printed: 05//8 Report cannot be used for OLCC/OHA compliance. Date Sampled: 05/07/8 00:00 Date Accepted:
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U N T E R S U C H U N G S B E F U N D
U N T E R S U C H U N G S B E F U N D Auftraggeber: schön & gut Katharina Schenke Lorettostrasse 30 79100 Freiburg Prüfsubstanz: Natives Olivenöl extra ex Griechenland (Ernte 2016 2017) ChargenBez.: 3517
Simplified Workflow and Improved Confidence in Screening Methods on Time of Flight Mass Spectrometry
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Batch 1. Batch 2. Batch 3. Batch 4. Batch 5
MACHEREY-NAGEL Chromatography Based on core-shell technology nenhanced π-π interactions and remarkable efficiency nexcellent stability in 00 % aqueous mobile phase nsuitable for LC/MS due to low bleeding
Waters Application Notes. Food Testing
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