1. Pr. Jean-Louis Habib Jiwan UCL – Département de chimie habib@chim.ucl.ac.be Les détecteurs de masse : une révolution en chromatographie 2ème partie : chromatographie et spectrométrie de masse

2. Couplings Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 EI CI ESI APCI APPI MALDI GC LC DESITLC On line Off line

3. Couplings : MALDI - LC Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Source : documentation Applied Biosystem LC MALDI - MS

4. On line couplings : acquisition modes Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 TIC RIC SIM SRM MRM ?

5. Acquisition modes and data treatment Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 time ● ● ● ● ● ● ● ● ● ● ● ● ● ● intensity

6. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 LC analysis of a pesticide mixture Acquisition modes and data treatment : pesticide mixture Source : B Ardrey « Liquid Chromatography-Mass Spectrometry. An Introduction », Wiley 2003 Micromass data

7. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Background mass spectrum obtained for this pesticide mixture analysis RIC Acquisition modes and data treatment : pesticide mixture

8. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes and data treatment : pesticide mixture Background substraction Component eluting after 4.65 min

9. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Background substraction Component eluting after 5.05 min Acquisition modes and data treatment : pesticide mixture

10. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes and data treatment : pesticide mixture

11. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes and data treatment : GC-MS / SCAN GC – MS of urine sample after trimethylsilylation

12. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes and data treatment : GC-MS / SCAN

13. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes and data treatment : Deconvolution GC – MS of hydrocarbon mixtures

14. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes and data treatment : METOCLOPRAMIDE LC-MS analysis APCI mode  Solid-state radiolysis of METOCLOPRAMIDE C14H22N3O2Cl Chemical mass 299.8 Exact mass 299.1400

15. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes : detection of target compounds SCAN, SIM or SRM ? SIM = Single Ion Monitoring SRM = Single Reaction Monitoring

16. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes and data treatment : Atrazine analysis R1R1 R2R2 R3R3 AtrazineC2H5C2H5 CH(CH 3 ) 2 Cl OHC2H5C2H5 CH(CH 3 ) 2 OH DEAHCH(CH 3 ) 2 Cl DIAC2H5C2H5 HCl CH 3 -CH=CH 2 MW = 42 [N=C-NH-C=N+H] + m/z = 68 LC-MS of Atrazine and its degradation products Full Scan Precursor ion scan m/z 68 Neutral loss scan 42 Da Source : RJ Steen et al, J Chromato A 2001, 915, 129

17. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes : SIM or SRM GC-MS of heptafluorobutyryl derivative of Methtryptoline RA Yost Adv Mass Spectrom 1985, 10B, 1479 SIM 362 m/z SRM 362 → 179 m/z

18. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes : Advanced methods on hybrid instruments FT ICR ORBITRAP TOF ION TRAP LC Chromatographic separation Ion source MS n capabilities HRMS Accurate mass capabilities

19. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acquisition modes : Advanced methods on hybrid instruments Source : documentation Shimadzu Naloxone Naltrexone 6-beta-naltrexol Buprenorphine MS 1 [M+H] + Expected 342.1705 MS 2 [342-H 2 O] + Expected 324.1600 MS 3 [324-C 4 H 6 ] + Expected 270.1113 LCMS – IT - TOF

20. LC - MS : quantitative analysis - glucosamine Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 D-Glucosamine chlorhydrate (C6H11O5NH2). HCl MW 215.683.1% of free base D-Glucosamine sulfate (C6H11O5NH2)2. H2SO4. 2KCl MW 605.5 59.2% of free base Difficulties : ► no chromophore ► highly polar ► positively charged at pH 7 HPLC Methods

21. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 HPLC Methods HILIC CH 3 CN / H 2 O (0.01% HCOOH) Mass spectrometry Ion Trap : MS 2 162 m/z (LCQ : LOD 25 ng/ml LOQ 50 ng/ml) Triple Quad : MS 2 neutral loss of 96 uma (TSQ : LOD 0.5 ng/ml LOQ 1 ng/ml) ESI mode  LC - MS : quantitative analysis - glucosamine

22. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 LC - MS : quantitative analysis - glucosamine LCQ results Tablets X : 298 mg/g (expected 443) Liquid Y : 0.7 mg/g Gel Z : 0.8 mg/g

23. TLC – MS : DESI Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Invention of Graham Cooks Science 2004, 306, 471 Source : Documentation from Prosolia Inc

24. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Source : G. Van Berkel et al, Anal. Chem. 2005, 77, 1207 TLC – MS : DESI

25. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 6G : R1,R2,R4 = CH2CH3; R3,R5 = H; R6, R7 = CH3 SRM m/z 443 → 415 B : R1, R6, R7 = H; R2, R3, R4, R5 = CH2CH3 SRM m/z 443 → 415 123 : R1 = CH3, R2,R3,R4,R5,R6,R7 = H SRM m/z 345 → 285 TLC: RP C2 Eluent : 80/20 MeOH/H2O 200 mM Ammonium acetate DESI solvent : MeOH TLC – MS : Rhodamines

26. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Source : G. Van Berkel et al, Anal. Chem. 2005, 77, 7183 TLC – MS : Rhodamines

27. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Caffeine Acetaminophen Aspirin TLC – MS : extra strength Excedrin extract TLC : NP eluent ACOEt/ACOH 99/1 DESI solvent : MeOH Full scan m/z 60 -300 [M+H] + = 195 m/z [M+Na] + = 217 m/Z [M+H] + = 152 m/z [M+Na] + = 174 m/z [M+Na] + = 203 m/z [M+2Na-H] + = 225 m/z

28. Pr. Jean-Louis Habib JiwanUCL – Département de chimieIPL mai 2007 Acknowledgments UCL, SC, CHIM, Mass Spec. Lab. Pr. Em. Ed de Hoffmann, A. Spôte, R. Rozenberg, R. Libert, L. Pesché UCL, MD, FARM, CHAM Pr. B. Tilquin, A. Maquille UCL, Saint-Luc Pr. Fr. Van Hoof, Pr. Ph. De Nayer €€€ : UCL FSR and COSIF