蔬菜、水果及制品中农兽药残留检测方案(其它食品专用)

智能文字提取功能测试中

TELEDYNE TEKMAREverywhereyoulookResidue in Apple Juice Using theAutoMate-Q40 Determination of Pesticide Application Note Abstract QuEChERS is a Quick-Easy-Cheap-Effective-Rugged-Safe extraction method that has been developed for thedetermination of pesticide residues in agricultural commodities. While the original unbuffered method wasdeveloped for plant matrices， since 2003， two additional buffered methods were created and adapted to manyadditional matrices such as fruit juices. The rise in popularity of the QuEChERS technique and the increase insample testing have driven the need for automation for this extraction technique. The AutoMate-Q40 streamlinesthe two part QuEChERS method from the liquid extraction to the cleanup step. The aim of this project is to evaluate the performance and versatility of the Automate-Q40.A LC-MS/MS was usedto determine pesticide residues in fruit juices， particularly in apple juice. Pesticide residues were extracted fromthe apple juice by using the AutoMate-Q40. Quantification was based on matrix-matched calibration curves withthe use of internal standard to ensure method accuracy.QC samples were evaluated at levels of 10， 50， 100 ng/gto ensure precision and accuracy of the AutoMate-Q40. Recent regulations on food analysis require screening for pesticides using confirmation techniques， such as LC-MS/MS. With the ever increasing amounts of pesticides being employed， 500 or more pesticides must be analyzedon a wide range of commodities. Even though the QuEChERS is a simplified extraction technique， it still requires many manual steps ranging fromaddition of solvent， extraction salts， centrifugation， shaking， decanting and performing the dSPE cleanup. Tomodernize the traditional QuEChERS extraction through the use of automation， Teledyne Tekmar has developedthe AutoMate-Q40. This automated platform will streamline the two part QuEChERS method from the liquidextraction through the cleanup. The goal of this work is to utilize the AutoMate-Q40 an automated QuEChERS extraction in a multi-lab validationstudy (Teledyne Tekmar and Pacific Agricultural Lab) for the determination of pesticides in Apple Juice. Pesticideresidues were extracted from the apple juice by using the AutoMate-Q40. Quantification was based on matrix-matched calibration curves with the use of internal standard to ensure method accuracy. QC samples wereevaluated at levels of 10， 50， 100 ng/g to ensure the precision and accuracy of the AutoMate-Q40. Procedure Sample Preparation/Extraction Apple juice samples were prepared according to the procedure described in the"AOAC Official Method 2007.01Pesticide Residues in Foods by Acetonitrile Extraction and Partitioning with Magnesium Sulfate". The sampleswere stored at room temperature and then stored in the refrigerator after opening. Figure 1 shows the sample preparation and extraction steps that are needed to extract the pesticide residues fromapple juice. For this analysis， the AutoMate-Q40 used AOAC QuEChERS extraction salts (MgSO4 and NaOAc). The AutoMate-Q40 also used the AOAC version of MgSO4， and PSA for the dSPE cleanup step. The following extractionwill be utilized in both analytical labs (Teledyne Tekmar and Pacific Agricultural Lab). Weigh 15.0+/-0.1g Apple Juice in 50mL centrifuge tube Place Samples into AutoMate-Q40 Add75uL of IS (TPP) solution and QC spike solution if necessary Add 15.0ml of ACN ADD 7.5g of AOAC QuEChERS extraction salt Cap and shake vigorously for 1.0 minute Centrifuge for 1.0 minutes Transfer 8.0mL to AOAC dSPE 15mL tube Shake for1 minute， centrifuge for 1 minutes Transfer 100.0uL extract and 900.0uL of Mobile Phase A to autosampler vial ， analyze by LC-MS/MS Figure 1： AutoMate-Q40 extraction parameters. Instrumentation and Analytical Conditions Two LC-MS/MS were run in Multiple Reactions Monitoring mode (MRM) for the initial Retention Time (RT) setup.Once the RT was established， the LC-MS/MS was switched over to Schedule Reaction Monitoring (SRM) for rest ofthe analysis. For confirmation of the extracted compounds， two transitions were determined for each compound.The mass transitions used are presented in Table 1， while the LC conditions are presented in Table 2. Lab A (Tekmar) AB Sciex 4500 QTrap Lab B (Pacific Agricultural Lab) Waters Xevo TQ-MS Compounds PrecursorIon (m/z) Quantization Product Ion (m/z) ConfirmationProduct Ion (m/z) Compounds Precursorlon(m/z) QuantizationProduct lon (m/z) confirmationProduct lon (m/z) Acetamiprid 222.93 125.90 98.90 Acetamiprid 223.00 56.10 126.00 Azoxystrobin 403.92 328.90 307.00 Azoxystrobin 404.10 329.00 372.00 Boscalid 344.80 307.00 139.00 Boscalid 342.90 139.90 307.00 Carbaryl 201.97 144.90 126.90 Carbaryl 202.00 127.00 145.00 Carbendazim 191.91 159.90 131.90 Carbendazim 192.00 160.00 132.00 Clofentezine 304.83 137.90 101.90 Clofentezine 303.00 102.00 138.00 Cyprodinil 225.93 93.00 108.10 Cyprodinil 226.00 93.00 108.00 Difenoconazole 405.79 250.80 187.90 Difenoconazole 406.00 251.10 111.10 Hexythiazox 352.90 227.70 167.90 Hexythiazox 353.00 168.00 228.00 Metconazole 320.01 124.90 98.90 Metconazole 320.10 70.00 125.00 Methoxyfenozide 369.02 148.80 91.00 Methoxyfenozide 369.10 149.10 313.20 Oxamyl 237.00 72.00 90.00 Oxamyl 237.00 72.00 90.00 Propiconazole 341.89 158.80 122.80 Propiconazole 342.00 159.00 69.00 Pyraclostrobin 387.89 193.90 162.80 Pyraclostrobin 388.10 163.00 193.90 Pyrimethanil 200.72 106.90 169.00 Pyrimethanil 200.00 82.00 107.00 Tebufenozide 353.03 132.90 296.90 Tebufenozide 353.10 133.00 297.10 Thiabendazole 201.86 174.90 131.00 Thiabendazole 202.00 131.00 175.00 Thiophanate Methyl 342.89 150.70 93.10 Thiophanate Methyl 343.00 93.00 151.00 Table 1： SRM transitions for LC-MS/MS parameters. Lab A (Teledyne Tekmar) Lab B (Pacific Agricultural Lab) Shimadzu Nexera LC Parameters Waters LC Parameters Column Restek Ultra AQ C18 Column Waters BEH C18 Dimensions 100 mm x 2.1 mm， 3 um Dimensions 2.1 mm x100 mm， 1.7 um Mobile Phase A 5mM Ammonium Formate in H，O Mobile Phase A：20mM Ammonium 0.2% acetic Acidin H20 B 5mM Ammonium Formate in MeOH B： 20mM Ammonium 0.2% acetic Acid in MeOH Gradient Time (min) %B Gradient Time (min) %B 0.10 10 Initial 30 1.50 10 2.00 30 4.00 60 8.00 50 8.00 70 14.00 90 11.00 100 17.00 90 12.00 100 18.00 30 12.01 10 15.00 10 Injection Vol (uL) 10.0 Injection Vol (uL) 10.0 Flow Rate (mL/min) 0.31 Flow Rate (mL/min) 0.45 Column Temperature (C) 30.0 Column Temperature (C) 60.0 Table 2： LC conditions parameters. Determination of Pesticide Experimental Results By automating the QuEChERS extraction， it enables a fast， easy， reliable and more reproducible extraction. TheAutoMate-Q40 offer significant labor savings， while it improves the repeatability and consistency between thesamples. A precision and accuracy study was performed in both labs using the AutoMate-Q40. A 6 ug/mL stock pesticidesolution was used to fortify the apple juice samples. Using the AutoMate-Q40， the system is able to automaticallyspike the samples with 25.0， 125.0 and 250.0 uL of the pesticide standard. These volumes produce a 10.0， 50.0 and100 ug/L check samples respectively. Also， the AutoMate-Q40 spiked in 75.0 uL of the internal standard TPP ineach sample to produce a concentration of 100.0 ug/L of. These QC samples were quantitated against theircorresponding matrix matched calibration curve. The analysis was performed in two parts. The AutoMate-Q40， can extract the apple juice samples with and withouta dSPE cleanup. Two sets of data will be presented in this precision and accuracy study.Table 3 shows data usingthe AutoMate-Q40 to extract unclean apple juice samples. While Table 4 shows data that was sent through thedSPE cleanup option using the AutoMate-Q40. Unclean Apple Juice Samples LAB A (Teledyne Tekmar) LAB B (Pacific Agricultural Lab) Compounds Low Spike Medium Spike High Spike Compounds Medium Spike High Spike % Recovery %RSD %Recovery %RSD %Recovery %RSD %Recovery %RSD %Recovery %RSD Acetamiprid 96.99 9.8 97.70 2.9 95.55 3.0 Acetamiprid 92.75 13.2 100.70 6.6 Azoxystrobin 104.09 2.9 99.90 3.4 101.13 1.7 Azoxystrobin 99.07 15.3 106.94 8.3 Boscalid 118.73 21.6 107.00 4.8 104.46 3.6 Boscalid 93.11 10.2 101.06 8.1 Carbaryl 106.86 3.5 103.95 2.8 103.33 3.4 Carbaryl 92.72 10.3 97.73 8.9 Carbendazim 98.02 2.7 100.63 2.4 96.60 6.5 Carbendazim 93.79 13.5 102.93 9.8 Clofentezine 104.73 8.3 101.37 3.2 102.24 4.0 Clofentezine 83.97 13.1 91.77 6.1 cyprodinil 102.93 6.2 101.13 2.2 102.27 5.6 Cyprodinil 91.43 11.3 95.43 4.9 Difenoconazole 105.14 2.6 95.22 2.0 98.83 3.5 Difenoconazole 95.43 10.0 104.46 2.6 Hexythiazox 113.82 9.8 97.63 2.6 97.79 4.9 Hexythiazox 90.18 10.4 93.66 3.2 Oxamyl 99.50 5.4 100.36 1.9 103.33 3.3 Oxamyl 86.36 15.9 93.87 7.4 Pyraclostrobin 90.13 7.9 98.77 3.1 98.82 5.3 Pyraclostrobin 81.32 13.5 88.31 8.0 Pyrimethanil 103.24 8.8 101.97 6.9 104.44 3.3 Pyrimethanil 92.39 11.5 102.44 4.4 Table 3： Unclean apple juice sample extracted using AutoMate-Q40 Cleaned Apple Juice Samples LAB A (Teledyne Tekmar) LAB B (Pacific Agricultural Lab) Compounds Low Spike Medium Spike High Spike Medium Spike High Spike % Recovery %RSD %Recovery %RSD %Recovery %RSD Compounds %Recovery %RSD %Recovery %RSD Acetamiprid 83.68 9.9 104.41 2.5 101.31 4.0 Acetamiprid 103.14 11.6 103.89 3.1 Azoxystrobin 101.53 7.2 104.66 2.7 102.21 4.5 Azoxystrobin 98.64 13.5 104.86 12.3 Boscalid 106.22 6.9 105.67 4.5 104.30 1.8 Boscalid 103.97 9.8 102.70 7.4 Carbaryl 104.09 7.4 105.57 2.0 104.51 2.9 Carbaryl 104.11 12.2 101.61 8.8 Carbendazim 93.74 11.8 101.89 3.5 93.36 6.2 Carbendazim 96.25 12.6 107.27 12.1 Clofentezine 98.06 6.6 102.26 2.8 99.69 3.2 Clofentezine 90.68 9.5 93.06 4.9 cyprodinil 104.61 5.2 106.10 3.9 104.03 3.2 Cyprodinil 97.61 11.1 101.16 4.3 Difenoconazole 102.79 9.0 99.73 3.1 96.79 5.0 Difenoconazole 103.50 9.6 108.14 2.5 Hexythiazox 102.65 7.6 100.49 3.2 99.06 4.9 Hexythiazox 87.25 8.8 91.44 4.4 Metconazole 98.22 8.8 102.58 4.0 98.96 5.0 Metconazole 98.22 12.9 105.61 4.2 Oxamyl 98.37 6.7 100.07 2.7 99.69 3.8 Oxamyl 93.82 17.0 87.59 7.5 Pyraclostrobin 88.02 16.5 102.36 6.8 98.51 6.1 Pyraclostrobin 87.54 8.5 99.23 4.1 Pyrimethanil 105.54 7.1 107.11 5.1 104.44 4.2 Pyrimethanil 95.68 6.2 106.67 8.3 Table 4： dSPE clean apple juice sample extracted using AutoMate-Q40 Table 3 and 4 shows that when using the AutoMate-Q40 to extract pesticide residues from apple juice， it exhibitsrecoveries ranging from 83.97% to 113.82% for the unclean samples， while the dSPE cleaned samples exhibitsexcellent recoveries ranging from 87.54% to 107.27%. These spike recoveries fall within the recommended meanvalues for the Document N° Sanco/12495/2011. This document states that the mean recoveries must fall within70% to 120% and a RSD <20%. The AutoMate-Q40，also， demonstrated great precision ranging from 1.7% to 21.6%RSD for the unclean samples and while the samples that used a dSPE cleanup showed precision ranging from2.0%to 17.0%RSD. This study demonstrates the feasibility of automating the QuEChERS extraction method using the AutoMate-Q40.By automating the liquid handing， addition of salt/buffers， sample mixing， pipetting， and liquid level sensing usingthe patent pending VialVisionTM， the extraction process is faster， more reliable， and easier. This enables time andlabor savings， while improving consistency and repeatability of the extraction. As shown above in Table 3 and 4 allpesticides gave excellent spike recoveries， ranging from 83.97% to 113.82% and excellent precision ranging from1.7% to 21.6%. Reference 1. European Committee for Standardization/Technical Committee CEN/TC275 (2008)， Foods of plantorigin： Determination of pesticide residues using GC-MS and/or LC-MS/MS following acetonitrileextraction/ partitioning and cleanup by dispersive SPE QuEChERS-method. 2. AOCA Official Method 2007.07 Pesticide Residues in Food by Acetonitrile Extraction and Partitioningwith Magnesium Sulfate. Gas Chromatography/Mass Spectrometry and Liquid Chromatography/TandemMass Spectrometry， First Action 2007 3. M. Anastassiades： QuEChERS a mini-multiresidue method for the analysis of pesticide residues in low-fat products 4. Method Validation and Quality Control Procedure for Pesticide Residues Analysis in Food and Feed(Document N° SANCO/12495/2011) Sales/Support：Main： Socialville Foster Rd.， Mason， OH etermination of Pesticide Residue in Apple Juice； -Aug-ww.teledynetekmar.com