The document discusses the implementation of a triple quadrupole mass spectrometry (TMS) system at BHH including its components and operating principles. It then provides details on the optimization and use of the system to develop LC-MS/MS methods for screening urine samples for drugs of abuse and developing steroid analysis services. The methods allow for the simultaneous detection of multiple analytes with high sensitivity and specificity.

1. Implmentation of TMS at BHH

3. General Principles

5. General Principles

7. Interface

8. General Principles

11. ESI Soft ionisation technique; little fragmentation Large molecules may be protonated more than once Peaks seen for same compound with different m/z ratios

16. General Principles

19. Mass Separation

20. General Principles

22. Electron Multiplier

24. TMS

31. MRM

32. TMS

45. Adducts and Clusters Formed in LC-MS Cluster/Adduct Source of Cluster Occurence Mass of Cluster Ion [M + CH3COO]- Acetic Acid APCI & IS -ve mode M + 59 [M + Cl]- Chlorinated Solvent APCI & IS -ve mode M + 35 [M + NH4]+ Ammonia APCI & IS +ve mode M + 18 [M + Na]+ Sodium Salts APCI & IS +ve mode M + 23 [M+K]+ Potassium Salts APCI & IS +ve mode M + 39 [M + CH3CNH]+ Acetonitrile APCI & IS +ve mode M + 42 [M+CH3OHH]+ Methanol APCI & IS +ve mode M + 33 [M+H3O]+ Water APCI & IS +ve mode M + 19

55. 152.1 300.2 Codeine 153 293.1 171.3 293.1 Benzoylcgonine-D3 181.2 292.2 152.1 292.2 Morphine-D6 150.1 290.1 168.1 290.1 Benzoylcgonine 198.2 290.1 154.1 290.1 Diazepam-D5 165.2 286.1 152.1 286.1 Morphine 192.9 285.1 154.1 285.1 Diazepam 180.2 282.2 236.2 282.2 Nitrazepam 249.1 278.2 234 278.2 EDDP 208.3 271.1 165.2 271.1 Nordiazepam 224.2 252.1 121.2 252.1 7-aminonitrazepam 119 150.1 91 150.1 Metamphetamine 66.9 142.2 93.1 142.2 125.1 142.2 Amphetamine-D6 119.2 136 91 136 Amphetamine Q3 Q1 Drug

56. Urine top std

57. Accuracy

61. Testosterone by LC-MS/MS (ESI)