! /!"#$!"#$%&'! !"#!"!"#$%&'!!"#$%&'()*+,-./0!"#$%&'()#*+,!"#$!"#$%&'(!"#$%&'!"#$%&'(!"#$%!"#!"#$%&'()*+,!"#$%&'()*+),-./
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1 ! /!"#$!"#$%&'!! Ralph Hindle Access Analytical Labs #3, Street N.E. Calgary, AB. T2E 7J8 Canada Agilent contact: Chin-Kai Meng Agilent Technologies, Inc. 285 Centerville Road Wilgton, DE USA!"!#$%&'()*+,-./ !"#$%&!"#$%&'(!"!"#$%&'()*+,-./!"#$5!"!"#$%&' 5 ppb (ng/g) 84%118%! 1-25 ppb!"#$%!"#$!"#$%&'(%!"#$!"#!"#$%&!"#$%&'()!"#$%&'()*+,-./1234!"#!"!"#$%&'!!"#$%&'()*+,-./!"#$%&'()#*+,!"#$!"#$%&'(!"#$%&'!"#$%&'(!"#$%!"#!"#$%&'()*+,!"#$%&'()*+),-./!"#$%&'()%&'*"+,-./1!"#$%&'!()*+(mrl) 1 ppb (ng/g) 1 ppb!"#$%&' MRL 1 ppb!"#$%&'()*+,-./!"#!"#$!"#$%&'!" ph!"#$%&!"#$!"#$(tlc)!!" Alberta!"#$% / (LC/MS)!"(APCI)!"#$%&!"!"#$%&'Alberta!"#$%&!"#$%&'()** +,-./!"#$%&
2 !"!#$%&!'()*+1!"!"#$%&'()*+,-./12!3!"#$%&'()[M+H] +!"#$!"#$%&'!"#$%&'(!"#$%&'()*+,-.!"#(CID)!"#$%&'()*+!"#$%&!"#$!"#$%&'()*Sigma Aldrich 99%!"# $%&'()*! 2 mg/ml!"#$% 3 ml.2n NaOH!"#$%&'(!"#$%&'()!"#$%&'( (IS)2 mg/ml!"#(scpd)!" HPLC!"#$%Caledon Labs (Georgetown, Ontario) (98%) EM Science!" 1 ml!" 1 µl 98%!! 8 mm!"# Ultra-Turrax T8!"# 5 ml!"#$13 mm!"#(pvdf)!"(.2 µm) VWR Scientific Oasis HLB (3 cc, 6 mg)!"#$ Waters! 1. 5 ml!"#$%&'( 3 g!" 2.!" 1 ml!" Ultra-Turrax 3 3.!"#$1!"#$%&'( 4.! 1 ml!"#$!"# 5.!"#!"#$%& 1 ml 6.!"# 1:4 25 µl + 75 µl.2 µm PVDF!"#$%!"# LC/MS!"#$%&'()*+,-./12!"#$%&'()*+,!"#$%!"#$%&'()*+,!"#$!"#$%&'()*+!"#$%&'()*!"#$%& SPE!"6 mg Oasis HLB! 1.5 ml!"#$ 1.5 ml!"#$1 ml!"#$%&' 1 ml!"# 1.5 ml!"#$%&'()*+!"#$% 1 ml 25%!"# LC/MS!"!"#$%1 mlspe!"#$%!"1 ml 25%!"#$!%&!"#$%&'() HPLC!"#$%!!"#$%&'(!"#$%&! 2
3 LC/MS LC/MS!"#$%&'( 11!"#!!"#!!"#$%&!"#$%&(MSD)!"#$%!"#"$%#&'()*+,"-.!"#!"#$%&'()*+! (MS)!"#$%&'()*+,-. LC!" APCI!"#$%&!'( [M+H] +!"#$%&'!"!"#$%& '()*+,!"!"#$%&'()!"#$%&'()!"#$%&'()*+,!"#$%!"#$!"#$%&'()*+!"#$%&!!"# / (GC/MS)!"!"# LC/MS/MS!"#$!"#$%&'(!"!"#$%&' LC/MS/MS 1. LC/MSD HPLC Zorbax Eclipse XDB-C8, 15 mm x 4.6 mm, 5 µm (P/N ) A.1%!" B.1%!" t = 2% B t1 = 2% B t3 = 9% B t6.5 = 9% B Post time = ml/!" 5 µl 3 MSD! APCI!"#$!"#$ 8!! 63 ms!" 7 V!" 6. L/!"# 6 psi!"#$ 35!"# 4!"# 3 V! 4 µa 3
4 Agilent 11 MSD!"#$%&'()* 4 MS!"#!"#$%&'()*!"#$%& 1!"#$%!"#$!"#$%&!"#$%!"#$%&[M+H] + 2!"#$%&'()*+!"#$%& 1 SIM!"#$%& 2!"#$%&'()*1! "#$% 1!"#$7 V (MSD1)22 V (MSD2) MSD1 25, EIC=249.7:25.7 (SULFAMSD\SSSCI1.D) APCI, Pos, SIM, Frag: 7, "Quantitation" SPY Simultaneous 2-signal aquisition; Fragmentor at 7 V or 2 V MSD1 285, EIC=284.7:285.7 (SULFAMSD\SSSCI1.D) APCI, Pos, SIM, Frag: 7, "Quantitation" SCPD (IS) MSD2 18, EIC=17.7:18.7 (SULFAMSD\SSSCI1.D) APCI, Pos, SIM, Frag: 2, "Confirmation" MSD2 156, EIC=155.7:156.7 (SULFAMSD\SSSCI1.D) APCI, Pos, SIM, Frag: 2, "Confirmation" MSD!! !"#$%&'()* 4
5 2!"#$%&'()*+,-./ !"#$%&'! (H 2 N + =[C 6 H 4 ]=O H 2 N + =[C 6 H 4 ]=SO 2!"#!"#$%&'(!"#$%&'( 2.!" APCI!"#$%&'(! (STZ) C 9 H 9 N 3 S 2 O 2 MW = 255 RT = 3.5 Fragmentor 7 V Fragmentor 16 V Fragmentor 2 V *MSD1 SPC, time=2.859:3.513 of SFCISCAN\SULFA6.D APCI, Pos, Scan, Frag: 7 1 Max: *MSD1 SPC, time=2.891:3.337 of SFCISCAN\SULFA8.D APCI, Pos, Scan, Frag: 16 1 Max: *MSD1 SPC, time=2.97:3.258 of SFCISCAN\SULFA9.D APCI, Pos, Scan, Frag: 2 Max: ! (SDZ) C 1 H 1 N 4 SO 2 MW = 25 RT = *MSD1 SPC, time=2.971:3.369 of SFCISCAN\SULFA1.D APCI, Pos, Scan, Frag: 7 Max: *MSD1 SPC, time=2.986:3.369 of SFCISCAN\SULFA12.D APCI, Pos, Scan, Frag: 16 1 Max: *MSD1 SPC, time=2.987:3.242 of SFCISCAN\SULFA13.D APCI, Pos, Scan, Frag: 2 Max: ! (SPY) C 11 H 11 N 3 SO 2 MW = 249 RT = *MSD1 SPC, time=3.212:3.467 of SFCISCAN\SULFA2.D APCI, Pos, Scan, Frag: 7 Max: *MSD1 SPC, time=3.21:3.68 of SFCISCAN\SULFA4.D APCI, Pos, Scan, Frag: 16 Max: *MSD1 SPC, time=3.21:3.529 of SFCISCAN\SULFA5.D APCI, Pos, Scan, Frag: 2 Max: *MSD1 SPC, time=3.626:3.865 of SFCISCAN\SULFA2.D APCI, Pos, Scan, Frag: 7 *MSD1 SPC, time=3.68:3.991 of SFCISCAN\SULFA4.D APCI, Pos, Scan, Frag: 16 *MSD1 SPC, time=3.672:3.927 of SFCISCAN\SULFA5.D APCI, Pos, Scan, Frag: 2!"# (SMR) C 11 H 12 N 4 SO 2 MW = 264 RT = Max: Max: Max:
6 2.!" APCI!"#$%&'( Fragmentor 7 V Fragmentor 16 V Fragmentor 2 V *MSD1 SPC, time=3.993:4.168 of SFCISCAN\SULFA2.D APCI, Pos, Scan, Frag: 7 1 Max: *MSD1 SPC, time=3.991:4.262 of SFCISCAN\SULFA4.D APCI, Pos, Scan, Frag: 16 1 Max: *MSD1 SPC, time=4.7:4.246 of SFCISCAN\SULFA5.D APCI, Pos, Scan, Frag: 2 1 Max: 1177!"#$ (SMZ) C 12 H 14 N 4 SO 2 MW = 278 RT = !" (SCPD) C 1 H 9 N 4 SO 2 CI MW = 284 RT = 4.31 *MSD1 SPC, time=4.184:4.423 of SFCISCAN\SULFA2.D APCI, Pos, Scan, Frag: 7 1 Max: *MSD1 SPC, time=4.262:4.453 of SFCISCAN\SULFA4.D APCI, Pos, Scan, Frag: 16 Max: *MSD1 SPC, time=4.262:4.438 of SFCISCAN\SULFA5.D APCI, Pos, Scan, Frag: 2 Max: *MSD1 SPC, time=4.47:4.725 of SFCISCAN\SULFA14.D APCI, Pos, Scan, Frag: 7 *MSD1 SPC, time=4.389:4.756 of SFCISCAN\SULFA16.D APCI, Pos, Scan, Frag: 16 *MSD1 SPC, time=4.469:4.772 of SFCISCAN\SULFA17.D APCI, Pos, Scan, Frag: 2! (SQ) C 14 H 12 N 4 SO 2 MW = 3 RT = Max: Max: Max: *MSD1 SPC, time=4.485:4.74 of SFCISCAN\SULFA18.D APCI, Pos, Scan, Frag: 7 *MSD1 SPC, time=4.454:4.9 of SFCISCAN\SULFA2.D APCI, Pos, Scan, Frag: 16 *MSD1 SPC, time=4.485:4.9 of SFCISCAN\SULFA21.D APCI, Pos, Scan, Frag: Max: Max: Max: 849!"#$% (SDMX) C 14 H 12 N 4 SO 2 MW = 31 RT =
7 !"#$%&'()*+,-./1!"#$%&'( SPY!" # 25!"!"# C 13!251!" 25!"#$%& SPY SDZ ( 251)!"# 2!"#$%&'()!"#$!"!"#$%&!"#$%& SDZ!"#$% SPY!"#$%!"#$%&'()*+ 2 1!"#$!"#$%!"#$%& 1.1!"#$%&'()*+,-!"!"#$%&'( )*+%,-!!"#$%&'()*!"#$!"#6!"#$%&'(!" 1! MSD1 256, EIC=255.7:256.7 (SULFAMSD\SSS_CI5.D) APCI, Pos, SIM, Frag: STZ MSD1 251, EIC=25.7:251.7 (SULFAMSD\SSS_CI5.D) APCI, Pos, SIM, Frag: SDZ MSD1 25, EIC=249.7:25.7 (SULFAMSD\SSS_CI5.D) APCI, Pos, SIM, Frag: SPY MSD1 265, EIC=264.7:265.7 (SULFAMSD\SSS_CI5.D) APCI, Pos, SIM, Frag: SMR MSD1 279, EIC=278.7:279.7 (SULFAMSD\SSS_CI5.D) APCI, Pos, SIM, Frag: SMZ MSD1 285, EIC=284.7:285.7 (SULFAMSD\SSS_CI5.D) APCI, Pos, SIM, Frag: SCPD (IS) MSD1 31, EIC=3.7:31.7 (SULFAMSD\SSS_CI5.D) APCI, Pos, SIM, Frag: 7 MSD1 311, EIC=31.7:311.7 (SULFAMSD\SSS_CI5.D) APCI, Pos, SIM, Frag: SQ SDMX 2.!"#$%!& 5 pg (SIM) 7
8 ! 3!"#(TIC)!"#$%&'()!"#$%&'()*+1 ml!"!" 25%!"#!"#$%!"#$%HPLC!" #$!"!"#$!"#$%&'!"#$!" MSD MSD!"#$%&'!"#$%&'!"#$%&'!"#$%&'()*+!"#$/!"#$(LC/MSD)!!!"#$%&'()*+,!"!!"#$ SPE!"#!!"#$%&'(!"#$%&'()!!"#$%&'()*+,-./1 3!"#$%&' SPE!"#$%&!"#$!"#$%&'()!" DL!"#$%&'()*+,- MSD1 TIC, MS File (SF3817\SULFA18.D) APCI, Pos, SIM, Frag: Spiked pork extract - solvent exchange to 25% MeOH in water MSD1 TIC, MS File (SF3816\SULFA11.D) APCI, Pos, SIM, Frag: 7 Spiked pork extract - Diluted 1 in 4 with water 6 MSD1 TIC, MS File (SF3817\SULFA1.D) APCI, Pos, SIM, Frag: 7 Spiked pork extract - Oasis HLB cleanup 6 3.!"#$%& TIC 8
9 !"#"$%&'()*+!"# MRL!"#$%&'()*+,!"#$%&'()*+ $,-.!!"#$% DL! 3!"#$%!"#$%&'(46!"#$% SIM!"#$%&' MSD1 256, EIC=255.7:256.7 (SF3817\SULFA18.D) APCI, Pos, SIM, Frag: STZ MSD1 251, EIC=25.7:251.7 (SF3817\SULFA18.D) APCI, Pos, SIM, Frag: SDZ MSD1 25, EIC=249.7:25.7 (SF3817\SULFA18.D) APCI, Pos, SIM, Frag: SPY MSD1 265, EIC=264.7:265.7 (SF3817\SULFA18.D) APCI, Pos, SIM, Frag: SMR MSD1 279, EIC=278.7:279.7 (SF3817\SULFA18.D) APCI, Pos, SIM, Frag: SMZ MSD1 285, EIC=284.7:285.7 (SF3817\SULFA18.D) APCI, Pos, SIM, Frag: SCPD (IS) MSD1 31, EIC=3.7:31.7 (SF3817\SULFA18.D) APCI, Pos, SIM, Frag: SQ MSD1 311, EIC=31.7:311.7 (SF3817\SULFA18.D) APCI, Pos, SIM, Frag: SDMX !"#(SIM) 9
10 MSD1 256, EIC=255.7:256.7 (SF3816\SULFA11.D) APCI, Pos, SIM, Frag: STZ MSD1 251, EIC=25.7:251.7 (SF3816\SULFA11.D) APCI, Pos, SIM, Frag: SDZ MSD1 25, EIC=249.7:25.7 (SF3816\SULFA11.D) APCI, Pos, SIM, Frag: SPY MSD1 265, EIC=264.7:265.7 (SF3816\SULFA11.D) APCI, Pos, SIM, Frag: SMR MSD1 279, EIC=278.7:279.7 (SF3816\SULFA11.D) APCI, Pos, SIM, Frag: SMZ MSD1 285, EIC=284.7:285.7 (SF3816\SULFA11.D) APCI, Pos, SIM, Frag: SCPD (IS) MSD1 31, EIC=3.7:31.7 (SF3816\SULFA11.D) APCI, Pos, SIM, Frag: SQ MSD1 311, EIC=31.7:311.7 (SF3816\SULFA11.D) APCI, Pos, SIM, Frag: SDMX 5. 1:4!
11 MSD1 256, EIC=255.7:256.7 (SF3817\SULFA1.D) APCI, Pos, SIM, Frag: STZ MSD1 251, EIC=25.7:251.7 (SF3817\SULFA1.D) APCI, Pos, SIM, Frag: SDZ MSD1 25, EIC=249.7:25.7 (SF3817\SULFA1.D) APCI, Pos, SIM, Frag: SPY MSD1 265, EIC=264.7:265.7 (SF3817\SULFA1.D) APCI, Pos, SIM, Frag: SMR MSD1 279, EIC=278.7:279.7 (SF3817\SULFA1.D) APCI, Pos, SIM, Frag: SMZ MSD1 285, EIC=284.7:285.7 (SF3817\SULFA1.D) APCI, Pos, SIM, Frag: SCPD (IS) MSD1 31, EIC=3.7:31.7 (SF3817\SULFA1.D) APCI, Pos, SIM, Frag: SQ MSD1 311, EIC=31.7:311.7 (SF3817\SULFA1.D) APCI, Pos, SIM, Frag: SDMX HLB (SIM)!" 7!"#$5 ppb 3 g!"#$ 15 ng!"#$%!"#$%&!"#$%& 3!"#$%& 3 ng SMR!"#$%&!"#!4!"#$%&3!84%-118%!"SPE!"#$%&'4!79%- 14%!"#$%SCPD!"5!!"#$%&'() 2 2 pg 2 pg SCPD!"#$% 7!"#!"#$%!"#$%&'()*+,-!"#$%&'()*+!"#$%&!"#$%&'(!(R 2 ) 34 11
12 3.!"#$% 1:4!"#$%&'( (ng) STZ SDZ SPY SMR SMZ SCPD(IS) SQ SDMX! , ! , ! , ! , ! , ! , ! , (ng) , , SD ( ) J 17 8 MDL (SDt-stat) ng J LOQ (SD1) ng J RSD (SD1/Mean) J 12 7 (%) (R 2 ) t-stat (N=7) !" Oasis HLB!"#$%&'( (ng) STZ SDZ SPY SMR SMZ SCPD(IS) SQ SDMX! , ! , ! , ! , ! , ! , ! , (ng) , , SD ( ) J 13 8 MDL (SDt-stat) ng J 4 26 LOQ (SD1) ng J RSD (SD1/Mean) J 1 7 (%) (R 2 ) t-stat (N=7)
13 5!"#$%! Oasis HLB!"#$!"!"!"#$%&'()*+!"#$!"#$%&'()*+, 1%!"!"#$%&'()*+!"!"#$%&'()HPLC!!!"#$%&!"#$!!"#$ HLB!"#$%&' ()!"#$%&'(MDL)!"#$%&'()*+,-. LC/APCI/MS!"#$%&'()*+,-./!"#$%&'()DL 125 ng/g SPE! 4 13 ng/g!!"#$%&'()*!"#$%&'!"# 1! 1. TLC-Densitometric Procedure for Sulfonamide Residues in Animal Tissue, SUL-SP8, Canadian Food Inspection Agency, Saskatoon, Saskatchewan, Canada; 21/4. 2. Sulfonamides in Tissue by LC/MS, Alberta Agriculture, Edmonton, Alberta, Canada, Standard Operating Procedure TX Mark Stahl, High-throughput analysis with the Agilent 11 Series high-throughput LC/MS system, Agilent Technologies, publication EN ! Oasis HLB!"#$%&'( STZ SDZ SPY SMR SMZ SCPD(IS) SQ SDMX % (1:4 ) SD ( ) J MDL (ng) J % (HLB ) SD ( ) J MDL (ng) J
14 CHCN
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