Solid Phase Extraction Of Fentanyl Analogues In Urine Using Nbe Hp-scx Columns On The Resolvex® I300.

Introduction

The detection of synthetic opioids including fentanyl in biological samples is crucial in clinical and forensic toxicology. Analysis techniques, such as liquid chromatography-tandem mass spectrometry (LC-MS/MS) enable multiplexed detection of fentanyl analogues with high selectivity and sensitivity. However, achieving high throughput with accurate quantification requires sophisticated sample preparation. Automated solid phase extraction (SPE) applied to fentanyl extraction with HP-SCX sorbent, offering reversed phase and strong cation exchange chemistry, addresses this challenge. Here, we highlight an automated workflow using the Resolvex^® i300 and HP-SCX Narrow Bore Extraction columns (NBE columns™) to extract 23 fentanyl analogues from human urine efficiently.

Sample preparation and SPE method.

25 fentanyl compounds (23 compounds and 2 internal standards) (Cerilliant Corp.) were utilized to demonstrate the HP-SCX SPE method. Compounds were diluted to 20 ng/mL, resulting in 1 ng/mL concentration when spiked into human urine samples. The Tecan Resolvex i300 was used for solvent dispensing, positive pressure loading, and analyte dry down. 24 technical replicate samples were processed using NBE columns with 5 mg of HP-SCX (30 μm) sorbent (Tecan P/N: 427-0051R-NBE). Utilizing the automated i300 workflow, 96 samples can be fully processed in 75 minutes.