Community Reference Laboratory for Pesticide Residues in Fruits and Vegetables. Amitraz Ring Test
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- Edwina Atkins
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2 Due to the concern in Europe with the use of amitraz in Turkey in pears, the CRL-SRM send a survey to know which laboratories where able to analyse amitraz. From this survey, it was proposed to the CRL-FV to perform a ring test to really be aware of the capacity the laboratories have. Invited participants were the countries involved in Turkish pears importation. These were the NRL-FV and the official laboratories belonging to these countries.
3 PARTICIPATION
4 PARTICIPATION United Kingdom Belgium The Netherlands Austria Romania Bulgaria Greece
5 PARTICIPATION In total 14 participants Country Austria Belgium Belgium Belgium Bulgaria Bulgaria Greece Greece Romania Romania The Netherlands United Kingdom United Kingdom NRL in FV yes yes no AO lab. yes yes yes no yes no yes no yes The CRL-SRM also participated
6 On the 27 th October, the CRL-FV, requested information to all the participating laboratories to ask for their needs on this ring test: - analytical standards. - analytical methods
7 So extraction methods for multiresidue method were sent by mail. On the 9 th November, 3 different analytical standard solutions of amitraz and its two main metabolites (DMPF and DMF) were sent by courier.
8 The CRL-FV sent a Protocol, a Calendar and reporting results: Form 1 and 2 ACTIVITY DATE Sample distribution. Deadline for receiving Form 1. Deadline for receiving results: Forms 2. Preliminary Report: only results. no statistical treatment. Final Report to the laboratories. 17 th November th November th November 2009 December 2009 January 2010
9 Blank samples were done from Alicante pears (Spain) The treated samples were two: Sample 1 Sample 2 Was treated ONLY with amitraz Was treated ONLY with the metabolites: DMF and DMPF One week was given to report results
10 Form 2: to give information on the concentration achieved and the methods used Form 1: to accept the sample
11 RESULTS Sample 1 only treated with amitraz Lab code Amitraz DMPF DMF Lab NA Lab11 NA 0.49 NA Lab Lab Lab NA Lab Lab Lab Lab Lab Lab Lab NA Lab06 Lab10 No Results Reported instrument failure - UK No Results Reported no method validation ready - Romania Median
12 RESULTS Sample 2 only treated with DMPF and DMF Lab code Amitraz DMPF DMF Lab NA Lab11 NA 0.09 NA Lab Lab Lab NA Lab Lab Lab Lab Lab Lab Lab03 NA Lab06 Lab10 No Results Reported instrument failure - UK No Results Reported no method validation ready - Romania Median
13 DEGRADATION ROUTE Me Me N C H N C H N Me Me Amitraz Me Me N C H H N Me Me H N CHO Me Me N-2.4-Dimethylphenyl-N-Methyl-formamidine 2.4- Dimethylformanilide DMPF Me NH 2 DMF Me 2.4-Dimethylanliline DMA
14 It has been applied two proposals* to determine amitraz overall concentration: 1) Amitraz gives DMPF and DMF in a 1:1 stereochemistry and expressed as amitraz. 2) Amitraz gives 2 molecules of DMA. Meanwhile, DMPF and DMF give each, one molecule of DMA and is expressed, the overall result, as DMA concentration. The results have been calculated using both ways *To be evaluated by the Scientific Advisory Group
15 1) Calculation as amitraz* Total C Amitraz + C DMPF Mw Amitraz + C DMF Mw = Amitraz Amitraz Mw DMPF Mw = DMF = C Amitraz + C DMPF C DMF 1.97 So applying this calculations to the results *considering an estichiometry 1:1
16 RESULTS Sample 1 only treated with amitraz Lab code Amitraz Total z-score Extraction Solvent Lab Acetone (MiniLuke) Lab Acetone (MiniLuke) Lab Acetone (MiniLuke) Lab Lab Lab Lab Lab Lab Lab Lab Lab Ethyl Acetate Median 0.476
17 5,0 4,0 3,0 2,0 1,0 0,0-1,0-2,0-3,0-4,0-5,0 Lab14 Lab07 Lab04 Lab02 Lab08 Lab13 Lab03 Lab12 Lab09 Lab01 Lab11 Lab05
18 RESULTS Sample 2 only treated with DMPF and DMF Lab code Amitraz Total z-score Extraction Solvent Lab Acetone (MiniLuke) Lab Acetone (MiniLuke) Lab Acetone (MiniLuke) Lab Lab Lab Lab Lab Lab Lab Lab Lab Ethyl Acetate Median 0.780
19 5,0 4,0 3,0 2,0 1,0 0,0-1,0-2,0-3,0-4,0-5,0 Lab09 Lab02 Lab11 Lab08 Lab04 Lab14 Lab03 Lab07 Lab13 Lab01 Lab12 Lab05
20 2) Calculation as DMA* Total C Amitraz Mw DMA + C DMPF Mw DMA + C DMF Mw = DMA = DMA 2 Mw Amitraz Mw DMPF Mw DMF = C Amitraz C DMPF C DMF So applying this calculations to the results * Considering a total conversion
21 RESULTS Sample 1 only treated with amitraz Lab code DMA Total z-score Extraction Solvent Lab09 0,213 0,5 Acetone (MiniLuke) Lab11 0,366 3,8 Acetone (MiniLuke) Lab14 0,190 0,0 Acetone (MiniLuke) Lab01 0,240 1,1 Lab02 0,189 0,0 Lab04 0,0922-2,0 Lab05 1, Lab07 0,0981-1,9 Lab08 0, Lab12 0, Lab13 0, Lab03 0, Ethyl Acetate Median 0.189
22 5,0 4,0 3,0 2,0 1,0 0,0-1,0-2,0-3,0-4,0-5,0 Lab04 Lab07 Lab03 Lab13 Lab08 Lab02 Lab14 Lab12 Lab09 Lab01 Lab11 Lab05
23 RESULTS Sample 2 only treated with DMPF and DMF Lab code DMA Total z-score Extraction Solvent Lab09 0,0299-3,6 Acetone (MiniLuke) Lab11 0,0672-3,2 Acetone (MiniLuke) Lab14 0,311-0,1 Acetone (MiniLuke) Lab01 0,379 0,7 Lab02 0,0306-3,6 Lab04 0,280-0,5 Lab05 0,506 2,3 Lab07 0,349 0,3 Lab08 0,270-0,6 Lab12 0,497 2,2 Lab13 0,351 0,4 Lab03 0,332 0,1 Ethyl Acetate Median 0.321
24 5,0 4,0 3,0 2,0 1,0 0,0-1,0-2,0-3,0-4,0-5,0 Lab09 Lab02 Lab11 Lab08 Lab04 Lab14 Lab03 Lab07 Lab13 Lab01 Lab12 Lab05
25 Conclusions: - The three Multirresidue methods applied (Miniluke, Ethyl Acetate and Quechers) could be effective to evaluate residues of Amitraz when LC- MS/MS is available - Depending on the extraction method applied, the DMPF and DMF contribution can vary, as a consequence of partial conversion between them. More information about this issue is needed. - A final statement about the conversion of DMF and DMPF in amitraz residue or DMA is important in order to harmonize the results obtained. - All NRL-FV from the border countries have implemented a method for the evaluation of Amitraz by MRM, although some of them have not achieved acceptable z-scores.
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Analysis of Ketamine and Xylazine in Rat Tissues Using the ACQUITY UPLC with 2D Technology Malorie Mella, 2 Brendan Schweitzer, 1 Sabra R. Botch-Jones, M.S., M.A, 1 Claude R. Mallet, Ph.D. 2 Boston University
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