Buprenorphine and norbuprenorphine determination in mice plasma and brain by gas chromatography-mass spectrometry

Abstract

A gas chromatography tandem mass spectrometry method for quantification of buprenorphine (BUP) and norbuprenorphine (NBUP) in brain and plasma samples from mice was developed and validated. Analytes were extracted from the brain or plasma by solid phase extraction and quantified within 20 minutes. Calibration was achieved by linear regression with a 1/x weighting factor and d4-buprenorphine internal standard. All products were linear from 1 to 2000 ng/mL with a correlation of determination >0.99. Assay accuracy and precision of back-calculated standards were within ±10%. The lower limit of quantification for both BUP and NBUP from the brain and plasma was 1 ng/mL. This sensitive and specific method can be used for the investigation of BUP mechanism of action and clinical profile.

Keywords: brain; buprenorphine; gc/ms; mice; norbuprenorphine; plasma.

Figure 1

Chemical structures of BUP, NBUP, and d₄-BUP.

Figure 2

Typical chromatogram and spectrum of BUP (200 ng/ml, A), NBUP (200 ng/ml, B), and d₄-BUP (C) spiked in plasma.

Figure 2

Typical chromatogram and spectrum of BUP (200 ng/ml, A), NBUP (200 ng/ml, B), and d₄-BUP (C) spiked in plasma.

Figure 2

Typical chromatogram and spectrum of BUP (200 ng/ml, A), NBUP (200 ng/ml, B), and d₄-BUP (C) spiked in plasma.

Figure 3

Representative chromatograms of plasma sample from a mice 30 minutes after intraperitoneal administration of 10 mg/kg BUP (A) and brain sample from a mice 30 minutes after cerebral perfusion of 1 mg/kg NBUP (B).

Figure 3

Representative chromatograms of plasma sample from a mice 30 minutes after intraperitoneal administration of 10 mg/kg BUP (A) and brain sample from a mice 30 minutes after cerebral perfusion of 1 mg/kg NBUP (B).