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L03:4:1971.  Application of liquid chromatography-tandem mass spectrometry to the analysis of benzodiazepines in blood

Rivera, H.M.(1), Walker, G.S.(1), Sims, D.N.(*2), Stockham, P.C.(2)

(1) Sch. Chem., Phys., Earth Sci., Flinders Univ., GPO Box 2100, Adelaide, SA 5001, (2) Forensic Sci. SA, 21 Divett Pl., Adelaide, SA 5000, Australia

REF: Eur. J. Mass Spectrom. 9(6) 2003 599-607

APCI LC/MS/MS is used for the simultaneous determination of 18 benzodiazepines and metabolites in human blood using model compds., then applied to real forensic cases incl. multiple drug overdose, sexual assaults and motor vehicle accidents. Blood (0.5 ml) is mixed with prazepam (int. std.) soln. then with excess 4.5% ammonia soln. and extd. with 1-chlorobutane. The solvent is reconstituted in 0.1 ml mobile phase.

LC: Agilent 1100, 5 µl inj., 15 cm x 4.6 mm i.d. (5 µm Zorbax XDB C₈), 60% MeOH in 20 mM ammonium formate (pH 9) for 15 min, to 100% MeOH over 1 min, hold 5 min, 0.7 ml/min.

MS: Agilent 1100 LC/MSD trap, APCI, pos. mode, corona 4000 nA, vaporizer 400°, skimmer 24 V, capillary exit 69 V, N₂ nebulizer gas 60 psi, drying gas 5 l/min (350°), CAD: anal. in 8 time segments, collision energy (optimized for each analyte) and transitions tabulated.

MS/MS spectra of all the analytes and typical ion chromatograms are reported. Therapeutic and sub-therapeutic levels of benzodiazepines are identified in real cases.

22 Refs.      4 Figs.      3 Tables

KEYWORDS: APCI/MS/MS, drugs of abuse; CAD; Ion trap; Benzodiazepines; Blood; Forensic;

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