Absorptive chemistry based extraction for LC-MS/MS analysis of small molecule analytes from biological fluids – an application for 25-hydroxyvitamin D

Abstract

The new TICE™ technology was found to be a useful process for sample preparation in clinical mass spectrometry. Full automation suited for routine analysis was achieved.

A simple micro-extraction plate assay for automated LC-MS/MS analysis of human serum 25-hydroxyvitamin D levels

“…In addition to losses during serum extraction, these recoveries also reflect analyte losses from ion suppression effects triggered by co‐eluting matrix compounds during ESI. The obtained recovery was 62% ± 8% RSD ( n = 3) in our experiments, which was lower than the 84% reported by Baecher et al for the same Tecan plates; these differences may be explained by inclusion of the ion suppression effects in the recovery rate here and the different HPLC method used by Baecher et al . Interestingly, in preliminary experiments, we found the micro‐extraction plates to be equally suited for the more hydrophilic vitamin D metabolites 1,25(OH) 2 D and 24,25(OH) 2 D, where we measured recovery rates of 60 and 65%, respectively (at 50 ng/ml).…”

“…All sample handling steps are therefore performed in a single well for each serum sample, which limits liquid transfer and pipetting steps, and also reduces potential sample losses. Pipetting and liquid mixing steps can be readily performed using a robotic autosampler with high accuracy and precision …”

“…Pipetting and liquid mixing steps can be readily performed using a robotic autosampler with high accuracy and precision. [6]…”

“…In vitamin D LC‐MS/MS assays, extraction from serum or plasma is usually performed using protein precipitation, liquid/liquid extraction (LLE), solid‐phase extraction (SPE) or combinations of these techniques . A novel extraction medium in 96‐well micro‐extraction plate format was recently introduced for lipophilic compounds such as vitamin D utilizing an immobilized sorptive material that combines all sample preparation steps on the same plate . This material links the advantages of LLE and SPE and requires only few sample processing steps for easy automation.…”

“…[4] A novel extraction medium in 96-well microextraction plate format was recently introduced for lipophilic compounds such as vitamin D utilizing an immobilized sorptive material that combines all sample preparation steps on the same plate. [6] This material links the advantages of LLE and SPE and requires only few sample processing steps for easy automation.…”