Extraction of Buprenorphine and Norbuprenorphine from Oral Fluid Using EVOLUTE® EXPRESS CX Prior to LC-MS/MS Analysis

Introduction

This application note describes a simple and robust mixedmode cation exchange solid phase extraction protocol using EVOLUTE^® EXPRESS CX 96-well plates for the extraction of buprenorphine and norbuprenorphine from oral fluid. Excellent extract cleanliness allows quantitation of the analytes across the range 0.1 ng/mL to 100 ng/mL using LC-MS/MS.

Figure 1. Structures of Buprenorphine and Norbuprenorphine.

EVOLUTE EXPRESS solid phase extraction products combine powerful EVOLUTE sorbent chemistry with enhanced ‘EXPRESS’ components. EVOLUTE EXPRESS products dramatically improve flow characteristics, and enhance sample preparation productivity. By truly eliminating the need for column conditioning and equilibration, samples can be prepared using a simple, fast load-wash-elute procedure. Analytes Buprenorphine, Norbuprenorphine Internal Standards Buprenorphine D₄, Norbuprenorphine D₃

Sample Preparation Procedure

Format: EVOLUTE^® EXPRESS CX 10 mg fixed well plate, part number 601-0010-PX01 Sample Pre-treatment Add 100 μL of sample (calibrator, QC or patient) to 900 μL of 0.1% aqueous formic acid. Either add an appropriate amount of internal standard separately or mix internal standard into formic acid pre-treatment solution prior to adding to sample. Sample Loading Load a 1 mL volume of pre-treated sample into each well at a flow rate of 1 mL/min (~10–12 drops/min).

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