Simultaneous Determination of Buprenorphine, Norbuprenorphine and Naloxone in Human Plasma by LC-MS-MS

Abstract

A novel sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS-MS) method simultaneously determined buprenorphine (BUP) and its active metabolite, norbuprenorphine (NBUP), and a coformulant, naloxone was developed, validated and applied successfully in humans. Buprenorphine-d 4 and norbuprenorphine-d 3 were used as the internal standard. The analysis was performed on a silica column, and the mobile phase was isocratic and composed of acetonitrile:2 mM ammonium formate in H2O (82:18, v/v). Mass spectrometry employed multiple reaction monitoring modes with transitions of m/z 468.1-55.2 for BUP, 414.2-101.2 for NBUP, 328.3-310.3 for naloxone, 472.1-59.2 for buprenorphine-d 4 and 417.2-101.2 for norbuprenorphine-d 3 . Lower limit of quantification (LLOQ) of the analytical method was 0.05 ng mL -1 for BUP, 0.1 ng mL -1 for NBUP and 0.025 ng mL -1 for naloxone, respectively. The standard calibration curves of BUP, NBUP and naloxone were linear over the concentration range of 0.05-20 ng mL -1 , 0.1-20 ng mL -1 and 0.025-20 ng mL -1 , respectively. The precisions (RSD) and accuracies (RE) of LLOQ and other QC samples were in acceptable range, with RSD \ 20% and RE ± 20% for LLOQ and RSD \ 15% and RE within ±15% for QC samples. The method was accurate, precise and specific, and was applied to the pharmacokinetic study of buprenorphine in healthy volunteers.

Molecularly imprinted electrochemical sensor prepared on a screen printed carbon electrode for naloxone detection

“…Several analytical methods have been reported for the determination of NLX mainly by high performance liquid chromatography (HPLC) [4] specially coupled with MS detection [1,[5][6][7][8][9]. Since NLX molecules can be easily oxidised, liquid chromatography with electrochemical detection was also used [10][11][12].…”

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

“…SPE (solid-phase extraction) was applied in many methods to clean up the plasma sample, which made the sample treatment highly cost and time consuming [23] . LLE (liquid–liquid extraction) was a simpler choice for sample treatment; however, larger sample volume together with large volume of extraction solvent was required, which was not suitable for green chemistry and made the procedures not practical in high-throughput analysis [24] .…”

“…Minimum effective concentration of the transdermal patch formulation of buprenorphine (Transtec ® ) was only 100 pg/mL in plasma and C max was only 305±117 pg/mL for 35 μg/h dose, which indicating the necessary for a highly sensitive method [6] . To date, liquid chromatography tandem mass spectrometry (LC-MS), with its advantages of high selectivity, sensitivity and accuracy, becomes a major technique for quantification of BUP and its metabolites in a wide variety of matrices, including urine [7] , [8] , [9] , [10] , [11] , [12] , [13] , [14] , [15] , plasma [16] , [17] , [18] , [19] , [20] , [21] , [22] , [23] , [24] , whole blood [9] , [10] , [11] , [25] , hair [9] , [10] , [11] , [26] , [27] , [28] , sweat [29] , meconium [30] , and breast milk [31] . Of the current available LC-MS methods applied in plasma, the lack of the requisite sensitivity and selectivity for accurate assessment of the drug concentrations is the major limitation.…”

“…A simpler, economic sample treatment procedure with excellent reproducibility and high recovery is also in need to meet the requirement of high-throughput analysis. [16] , [17] , [18] , [21] , [24] .…”

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

“…These approaches include radioimmunoassay [ 10 ], ELISA assay [ 11 ], cloned enzyme donor immunoassay (CEDIA) [ 12 ], gas chromatography coupled with mass spectrometry [ 13 , 14 , 15 ], as well as liquid chromatography coupled with UV [ 16 ] or fluorescent detectors [ 17 ]. Methods based on liquid chromatography and mass spectrometry [ 18 , 19 ] or tandem mass spectrometry [ 20 , 21 , 22 , 23 , 24 ] seem to provide the highest sensitivity. Nevertheless, almost uniformly the main issue reported by the authors was associated with achieving a satisfactory level of sensitivity.…”