A sensitive, simple and rapid HPLCâMS/MS method for simultaneous quantification of buprenorpine and its N-dealkylated metabolite norbuprenorphine in human plasma

• Published in: Journal of pharmaceutical analysis Vol. 3; no. 4; pp. 221 - 228
• Main Authors: Yi-Ya Wang, Xiao-Hang Shen, Hao Li, Feng-Ju Chen, Yan Fu, Li Ding
• Format: Journal Article
• Language: English
• Published: Elsevier 01.08.2013
• ISSN: 2095-1779

A sensitive, simple and rapid high performance liquid chromatography-tandem mass spectrometry (HPLCâMS/MS) method was developed and fully validated for the simultaneous quantification of buprenorphine (BUP) and its N-dealkylated metabolite norbuprenorphine (NBUP) in 200μL human plasma. Human plasma samples were prepared using liquidâliquid extraction, and then separated on a Shiseido MG C18 (5μm, 2.0mmÃ50mm) via 4.1min gradient elution. Following electrospray ionization, the analytes were quantified on a tripleâquadrupole mass spectrometer in multiple-reaction-monitoring (MRM) positive ion mode. Linearity was achieved from 25.0 to 10000pg/mL for buprenorphine, from 20.0 to 8000pg/mL for norbuprenorphine with r2>0.99. The method was demonstrated with acceptable accuracy, precision and specificity for the detection of buprenorphine and norbuprenorphine. Recovery was 81.8â88.8% for buprenorphine and 77.0â84.6% for norbuprenorphine, and the matrix effect was 95.6â97.4% for buprenorphine and 94.0â96.9% for norbuprenorphine; all were not concentration dependent. With validated matrix and autosampler stability data, this method was successfully applied in a bioequivalence study to support abbreviated new drug application. Keywords: Buprenorphine, Norbuprenorphine, Human plasma, HPLCâMS/MS