Conclusions
The isotopically labelled internal standard UPLC-MS/MS method is established and well applied in the clinical setting. A strong correlation but discordance was found between UPLC-MS/MS and EMIT, indicating that switching from UPLC-MS/MS to EMIT was unsuitable. However, the adjusted EMIT results may serve as a reliable surrogate when UPLC-MS/MS results cannot be obtained when necessary.
Methods
An isotopically labelled internal standard UPLC-MS/MS method was established, validated, and subsequently applied to determine VRZ concentration. The UPLC-MS/MS method was also compared with a commercial EMIT method regarding
Results
The calibration curve of UPLC-MS/MS was linear from 0.1 to 10 mg/L, the inter- and intra-day relative standard deviations (RSDs), and the stability of quality control samples were less than 15 %, satisfying the Bioanalytical Method Validation Guidelines. A total of 122 plasma samples were collected and analyzed using both methods. UPLC-MS/MS and EMIT showed a high correlation (r = 0.9534), and Bland-Altman analysis indicated a mean absolute bias of 1.035 mg/L and an average bias of 27.56 % between UPLC-MS/MS and EMIT. The paired Wilcoxon test and Bland-Altman analysis revealed poor consistency between the two methods. Furthermore, we compared the effects of different methods in clinical applications. Two threshold values for treatment efficacy (1.0 mg/L) and safety (5.5 mg/L) were established, and considerable discordance was observed between the original EMIT and UPLC-MS/MS results at both thresholds (p < 0.05). Nevertheless, the adjusted EMIT results were not inconsistent with the UPLC-MS/MS results regarding the efficacy (p = 0.125) and safety (p = 1.0) thresholds. Conclusions: The isotopically labelled internal standard UPLC-MS/MS method is established and well applied in the clinical setting. A strong correlation but discordance was found between UPLC-MS/MS and EMIT, indicating that switching from UPLC-MS/MS to EMIT was unsuitable. However, the adjusted EMIT results may serve as a reliable surrogate when UPLC-MS/MS results cannot be obtained when necessary.