Quantitative Analysis of Alprazolam and Triazolam in Hemolysed Whole Blood and Liver Digest by GC/MS/NICI with Deuterated Internal Standards

This report describes sensitive and specific methods for the quantitation of alprazolam and triazolam in hemolysed whole blood and liver tissue. Samples of blood and enzyme-digested liver are extracted without pH adjustment with n-butyl chloride, after addition of deuterated internal standards and urea. The evaporated extracts are reconstituted in acetonitrile for analysis by gas chromatography/mass spectrometry/negative ion chemical ionization (GC/MS/NICI). Fatty extracts may be cleaned up by partitioning between pentane and acetonitrile. Two ion pairs are monitored for each drug. Within-day coefficients of variation in the range 10–50 µg/L for blood are approximately 5%. Between-day coefficients of variation are less than 10%. The limit of quantitation (based on analysis of 0.2-mL blood samples) is 0.5 µg/L for triazolam and 4 µg/L for alprazolam.