Performance evaluation of automated magnetic beads extraction method for the measurement of 23 plasma steroids using...

To evaluate the performance of magnetic beads extraction method (MGE) for quantifying plasma steroids using liquid chromatography tandem mass spectrometry (LC-MS/MS).

The evaluation encompassed calibration linearity, limit of detection (LOD), lower limit of the measuring interval (LLMI), specificity and matrix effects, trueness and recovery, intra- and inter-day precisions, repeatability, dilution consistency and stability of 23 plasma steroids extracted by MGE method were evaluated. The 23 plasma steroids were isolated and analyzed through a single sample preparation and two injections by LC-MS/MS.

All 23 steroids were successfully resolved chromatographically within 12.1 min. The automated MGE method exhibited excellent linearity (R² > 0.995 for all analytes) with coefficients of variation (CVs) ranging from 3.36% to 9.28% at LLMI and from 1.58% to 16.53% at LOD. Additionally, the CVs and average deviation for repeatability were between 0.55% and 7.11%, and - 6.57% to 10.81%, respectively. Both intra-day precision (0.88% to 8.43%) and inter-day precision (0.98% to 6.51%) satisfied the acceptance criteria. The average deviation for trueness and recovery ranged from -13.93% to 9.06%. Moreover, specificity, matrix effect, dilution consistency, and stability were distinctly identified and conformed to guideline requirements. Notably, levels of AD, T, E1, and 17-OHP were significantly elevated in patients with polycystic ovary syndrome (PCOS) (p < 0.05).

The automated MGE method demonstrates high efficiency and reliability for the simultaneous quantification of 23 plasma steroids, offering a promising solution for high-throughput analysis of steroid panels in clinical in the future.