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

Abstract

A sensitive, simple and rapid high performance liquid chromatography-tandem mass spectrometry (HPLC–MS/MS) method was developed and fully validated for the simultaneous quantification of buprenorphine (BUP) and its N-dealkylated metabolite norbuprenorphine (NBUP) in 200 μL human plasma. Human plasma samples were prepared using liquid–liquid extraction, and then separated on a Shiseido MG C18 (5 μm, 2.0 mm×50 mm) via 4.1 min gradient elution. Following electrospray ionization, the analytes were quantified on a triple–quadrupole mass spectrometer in multiple-reaction-monitoring (MRM) positive ion mode. Linearity was achieved from 25.0 to 10000 pg/mL for buprenorphine, from 20.0 to 8000 pg/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 81.8–88.8% for buprenorphine and 77.0–84.6% for norbuprenorphine, and the matrix effect was 95.6–97.4% for buprenorphine and 94.0–96.9% 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.

Simultaneous determination and pharmacokinetic study of Atractylenolide I, II and III in rat plasma after intragastric administration of Baizhufuling extract and Atractylodis extract by UPLC–MS/MS

“…Solid phase extraction (SPE) was applied in many methods to clean up the plasma sample, which made the sample treatment highly cost and time consuming [25]. Several sample pre-treatment methods were investigated in this study including protein precipitation and LLE.…”

High-Performance Liquid Chromatography-Tandem Mass Spectrometry for Buprenorphine Evaluation in Plasma—Application to Pharmacokinetic Studies in Rabbits

“…BUP is a polar compound, therefore the electrospray ionization (ESI) mode was selected for the analysis [ 21 , 23 , 25 , 27 , 29 , 30 , 31 ]. All the parameters (declustering potential (DP), collision energy (CE), entrance potential) have been optimized with a direct infusion of working standard solutions ( Table 1 ).…”

“…All the parameters (declustering potential (DP), collision energy (CE), entrance potential) have been optimized with a direct infusion of working standard solutions ( Table 1 ). Many authors pointed out the problem of the quality and stability of BUP ionization at low CEs (below 30 eV) [ 21 , 23 , 25 , 29 , 31 ]. To overcome that issue, one team [ 31 ] included trifluoroacetate ammonium in the mobile phase and obtained an improved ionization stability.…”

“…Many authors pointed out the problem of the quality and stability of BUP ionization at low CEs (below 30 eV) [ 21 , 23 , 25 , 29 , 31 ]. To overcome that issue, one team [ 31 ] included trifluoroacetate ammonium in the mobile phase and obtained an improved ionization stability. Other authors monitored the precursor ion (468.6) as the highest intensity ion [ 21 , 23 , 25 , 29 ].…”

“…Most authors reported C18 columns as a stationary phase in BUP determination [ 23 , 25 , 27 , 29 , 30 , 31 ]. An alternative approach was proposed by Favretto et al [ 21 ].…”