|SOFT - TIAFT 1998||Poster Session 4||Friday October 9, 1998|
HPLC/MS/MS ANALYSIS OF FLUNITRAZEPAM, METABOLITES, AND OTHER BENZODIAZEPINES IN URINE AND WHOLE BLOOD|
Robert K. Lantz, Patricia L.Sulik, and Brian J. Kirby
Rocky Mountain Instrumental Laboratories, 108 Coronado Ct., Ft. Collins, CO 80526, USA
|Flunitrazepam (FNP), des-methyl-flunitrazepam, 7-amino-flunitrazepam, and 7-acetamido-flunitrazepam are analyzed by HPLC/MS/MS using a Waters C18 Symmetry column (3.9mm x 150mm), ammonium acetate buffer (pH 4.8, 40mM acetate, 40% v/v acetonitrile), 1.0mL/min. The internal standard is d5-nordiazepam. MS instrumentation includes Shimadzu LC10A/d pumps, P-E SciEx API-300 triple quadrupole mass spectrometer run in the MS/Ms mode, with SciEx MassChrom quantitation software. Ion transitions used are as follows: flunitrazepam (FNP): 314.2/205.1; 7-amino-FNP: 284.1/227.1; des-methyl-FNP: 300.1/254.1; 3-hydroxy-FNP: 330.1/284.1; 7-acetamido-FNP: 326.2/227.2; d5-nordiazepam: 276.1/213.0. |
Blood, standards, and hydrolyzed urine samples are mixed with internal standard, then extracted with n-butyl chloride. The organic layer is separated and dried under nitrogen. The organic layer is then reconstituted with mobile phase and analyzed in duplicate.
LOD is less than 1 ng/mL for 14 benzodiazepines and metabolites tested. LOQ is 2 ng/mL for the same group of benzodiazepines and metabolites.