Lykkeberg, Anne Kruse

Poulsen, Mette Erecius

Online Research Database In Technology

                       General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.  • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal   If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.     Downloaded from orbit.dtu.dk on: Dec 20, 2017Application of HR-LC-MS in screening methods for pesticides in cerealsLykkeberg, Anne Kruse; Poulsen, Mette EreciusPublication date:2014Document VersionPeer reviewed versionLink back to DTU OrbitCitation (APA):Lykkeberg, A. K., & Poulsen, M. E. (2014). Application of HR-LC-MS in screening methods for pesticides incereals [Sound/Visual production (digital)]. Joint Training EURL-CF/FV LC and GC HRMS, Almeria, Spain,12/06/2014Application of HR-LC-MS in screening methods for pesticides in cereals Anne Kruse Lykkeberg, Mette Erecius Poulsen Almeria, 12 June 2014 National Food Institute, Technical University of Denmark Disposition Analysis at the instrument Standard mixtures Validation plan Validation Virtual computers – for data treatment Sample preparation of cereals – QuEChERS National Food Institute, Technical University of Denmark LC-QTOF instruments National Food Institute, Technical University of Denmark Disposition Analysis at the instrument Standard mixtures Validation plan Validation Virtual computers – for data treatment Sample preparation of cereals – QuEChERS National Food Institute, Technical University of Denmark PCD – CSV file National Food Institute, Technical University of Denmark No selection  QTOF-analysis – MS scan No collision  • Used to determine retention time  National Food Institute, Technical University of Denmark PCDL - library National Food Institute, Technical University of Denmark PCDL – library - spectra National Food Institute, Technical University of Denmark Selection of single mass QTOF-analysis – Targeted MS/MS Collision 10, 20 and 40 V  • Obtaining compound spectra for Library • Spectra of daughter ions from the single ion isolated in the quadropole  National Food Institute, Technical University of Denmark No selection QTOF-analysis – All ion MS  Collision 10, 20 and 40 V  • MS scan with collision energy • Screening samples • Spectra of daughter ions of all compounds eluting on a certain time point  National Food Institute, Technical University of Denmark Chromatogram - TIC National Food Institute, Technical University of Denmark Spectra – MSscan incl. collision 0V 10V 20V 40V Ofurace, [M+H]+ National Food Institute, Technical University of Denmark Disposition Analysis at the instrument Standard mixtures Validation plan Validation Virtual computers – for data treatment Sample preparation of cereals – QuEChERS National Food Institute, Technical University of Denmark Standard mixtures 50-100 pesticides Mixtures Avoiding isomers in the mixtures Isomers 2-6 mixtures together Validation National Food Institute, Technical University of Denmark Spectra of isomers in Agilents library  Library spectrumm/z30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250Abundance0102030405060708090100110142.07234100.00 184.1192996.84226.1662467.7486.0348915.20100.0505412.1068.024324.7957.044724.62152 118231 62 Library spectrumm/z30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250Abundance0102030405060708090100110170.10364100.00226.166246.72114.066185.83 142.072345.31100.050543.8275.055293.1185.076032.24 128.081832.1057 069881 58 Library spectrumm/z30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250Abundance0102030405060708090100110170.10364100.00226.1662441.95100.0505410.48 114.066189.97 142.072349.5875.055294.96 86.034894.38 128.081834.1257.069882.73NNNNHNH CH3CH3CH3CH3OCH3NNNNHNH CH3CH3CH3OCH3CH3NNNNHNHCH3CH3OCH3CH3Prometon Terbumeton Secbumeton National Food Institute, Technical University of Denmark Disposition Analysis at the instrument Standard mixtures Validation plan Validation Virtual computers – for data treatment Sample preparation of cereals – QuEChERS National Food Institute, Technical University of Denmark Validation • My alternative validation plan • 20-25 samples – spiked at 0.1 mg/kg • 10-20 blank samples     Spiked – 0.1 mg/kg Blank Rice 5 5 Oat 5 5 Barley 5 5 Wheat 5 5 Rye 5 5 National Food Institute, Technical University of Denmark Dilution of spiked samples         Spiked rye is diluted with Blank rye …  Matrix is the same Recovery is assumed to be the same at the different concentration levels  Rye 0.1 mg/kg Blank  0.10 mg/kg 1000 µl - 0.02 mg/kg 200 µl 800 µl 0.01 mg/kg 100 µl 900 µl Blank - 1000 µl National Food Institute, Technical University of Denmark Disposition Analysis at the instrument Standard mixtures Validation plan Validation Virtual computers – for data treatment Sample preparation of cereals – QuEChERS National Food Institute, Technical University of Denmark Compound list – exported to Excel National Food Institute, Technical University of Denmark All file gathered in one Excel-file National Food Institute, Technical University of Denmark Area of all samples spiked at 10 ppb National Food Institute, Technical University of Denmark Comparison of areas in spiked samples with blank National Food Institute, Technical University of Denmark Disposition Analysis at the instrument Standard mixtures Validation plan Validation Virtual computers – for data treatment Sample preparation of cereals – QuEChERS National Food Institute, Technical University of Denmark Virtual computers for data-analysis Challenges • The Agilent software demands Windows Professional 64 bit – english version • Back up of all laboratory computer on LAB-domain • Laboratory domain, with no access from personal computers Solution • Virtual computers from Citrix –On the Laboratory domain –Windows Professional 64 bit –Access to all data – back up early in the morning –Few licenses are necessary –Access from all computers with internet National Food Institute, Technical University of Denmark Disposition Analysis at the instrument Standard mixtures Validation plan Validation Virtual computers – for data treatment Sample preparation of cereals – QuEChERS National Food Institute, Technical University of Denmark FVII cocktail IV – project  • Aim is to develop a cereal-method to analyze many analytes in one method.  • Analytes: – Pesticides (The pesticides most present in cereal – in practice the standard mixtures which we use for our quantitative methods).  – Mycotoxins: Most important mycotoxins in cereals.   National Food Institute, Technical University of Denmark Method development - QuEChERS •Water • Acetonitrile Extraction •Magnesium sulphate •NaCl, Citrate buffer Phase separation •1 h at -80 ºC • Centrifugation Freeze out step • PSA  •Magnesium sulphate PSA clean up National Food Institute, Technical University of Denmark 020040060080010001200140016000 2 4 6 8 10 12 14 16 18 20m/z Retention time Positive mode Pos blank 1Extraction National Food Institute, Technical University of Denmark 020040060080010001200140016000 2 4 6 8 10 12 14 16 18 20m/z Retention time Positive mode Pos blank 1Pos blank 2Phase separation National Food Institute, Technical University of Denmark 020040060080010001200140016000 2 4 6 8 10 12 14 16 18 20m/z Retention time Positive mode Pos blank 1Pos blank 2Pos blank 3Freeze out step National Food Institute, Technical University of Denmark 020040060080010001200140016000 2 4 6 8 10 12 14 16 18 20m/z Retention time Positive mode Pos blank 1Pos blank 2Pos blank 3Pos blank 4PSA clean up National Food Institute, Technical University of Denmark           020040060080010001200140016000 2 4 6 8 10 12 14 16 18 20m/z Retention time Positive mode Pos blank 1Pos blank 2Pos blank 3Pos blank 4MycotoxinsPesticidesAnalytes National Food Institute, Technical University of Denmark 01002003004005006007008000 2 4 6 8 10 12 14 16 18m/z Retention time Negative mode Neg blank 1Extraction National Food Institute, Technical University of Denmark 01002003004005006007008000 2 4 6 8 10 12 14 16 18m/z Retention time Negative mode Neg blank 1Neg blank 2Phase separation National Food Institute, Technical University of Denmark 01002003004005006007008000 2 4 6 8 10 12 14 16 18m/z Retention time Negative mode Neg blank 1Neg blank 2Neg blank 3Freeze out step National Food Institute, Technical University of Denmark 01002003004005006007008000 2 4 6 8 10 12 14 16 18m/z Retention time Negative mode Neg blank 1Neg blank 2Neg blank 3Neg blank 4PSA clean up National Food Institute, Technical University of Denmark 01002003004005006007008000 2 4 6 8 10 12 14 16 18m/z Retention time Negative mode Neg blank 1Neg blank 2Neg blank 3Neg blank 4MycotoxinsPesticidesAnalytes National Food Institute, Technical University of Denmark TIC af blank i positiv mode 7x100000011111222223333344444.55.5.+ESI TIC Scan pos_Blank4.d 2Counts vs. Acquisition Time (min)1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14 14.5 15 15.5 16 16.5 17 17.5 18National Food Institute, Technical University of Denmark TIC af blank i positiv mode 7x100000011111222223333344444.55.5.+ESI TIC Scan pos_Blank4.d Counts vs. Acquisition Time (min)6.6 6.8 7 7.2 7.4 7.6 7.8 8 8.2 8.4 8.6 8.8 9 9.2 9.4 9.6 9.8 10 10.2 10.4 10.6 10.8 11 11.2 11.4 11.6 11.8 12 12.2 12.4 12.6 12.8National Food Institute, Technical University of Denmark TIC af blank i negativ mode 7x1000000000001111111111222222.2.-ESI TIC Scan neg_Blank4.d 1 12Counts vs. Acquisition Time (min)0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14 14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 19.5National Food Institute, Technical University of Denmark TIC af blank i negativ mode 7x1000000000001111111111222222.2.-ESI TIC Scan neg_Blank4.d Counts vs. Acquisition Time (min)6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8 8 8.2 8.4 8.6 8.8 9 9.2 9.4 9.6 9.8 10 10.2 10.4 10.6 10.8 11 11.2 11.4 11.6 11.8 12 12.2 12.4 12.6 12.8 13National Food Institute, Technical University of Denmark Matrix effect & recovery – Mycophenol acid • Mycophenol acid • 6.63 min • Mass: 320.1260 Recovery Matrix effect PSA 7 106 Freeze out 114 153 Phase separation 105 149 Extraction 123 159 National Food Institute, Technical University of Denmark Matrix effect & recovery – Fumonisin B2 • Fumonisin B2 • 5.07 min  • Mass: 705.3936 Recovery Matrix effect PSA 6 111 Freeze out 36 151 Phase separation 73 166 Extraction 76 153 National Food Institute, Technical University of Denmark Matrix effect & recovery – Roquefortin C • Roquefortin C • 5.15 min  • Mass: 389.1852 Recovery Matrix effect PSA 92 75 Freeze out 92 41 Phase separation 101 38 Extraction 153 31 National Food Institute, Technical University of Denmark Matrix effect & recovery – Clothianidin • Clothianidin • 4.46 min  • Mass: 249.0087 Recovery Matrix effect PSA 89 83 Freeze out 95 61 Phase separation 76 57 Extraction 153 32 

Application of HR-LC-MS in screening methods for pesticides in cereals

http://orbit.dtu.dk/ws/files/102669784/LC_QTOF_Almeria.pdf

Application of HR-LC-MS in screening methods for pesticides in cereals

Abstract

Similar works

Full text

Available Versions

Online Research Database In Technology