Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510, Japan.
Anal Sci. 2020 Sep 10;36(9):1099-1104. doi: 10.2116/analsci.20P082. Epub 2020 Apr 24.
Liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) enables the accurate and precise quantification of various analytes at very low concentrations, but it has room for improvement in analysis throughput. Multiplexing of samples in the same injection could be a promising procedure for enhancing the analysis throughput. This could be achieved by derivatization of the multiple samples with multiple isotopologous reagents. In this study, a sample-multiplexed LC/ESI-MS/MS assay using the 1-[(4-dimethylaminophenyl)carbonyl]piperazine (DAPPZ) isotopologues (H-, H-, and H-forms) was developed and validated for the simultaneous determination of primary bile acids in three different plasma samples in a single run. The developed method had satisfactory intra- and inter-assay precisions (≤ 2.3 and ≤ 4.2%, respectively) and accuracy (99.0 - 100.3%), and could reduce the total LC/ESI-MS/MS run time by more than 60% for 42 samples compared to the conventional method. Thus, the derivatization with the DAPPZ isotopologues worked well for enhancing the throughput of the LC/ESI-MS/MS assay of the bile acids.
液相色谱/电喷雾串联质谱（LC/ESI-MS/MS）能够准确、精确地定量检测极低浓度的各种分析物，但在分析通量方面仍有改进的空间。在同一次进样中对多个样品进行多重化处理可能是提高分析通量的一种很有前途的方法。这可以通过用多个同位素类似物对多个样品进行衍生化来实现。本研究开发并验证了一种使用 1-[(4-二甲氨基苯基)羰基]哌嗪（DAPPZ）同位素（H-、H-和 H-形式）的样品多重化 LC/ESI-MS/MS 测定法，可在单次运行中同时测定三种不同血浆样品中的初级胆汁酸。所开发的方法具有令人满意的日内和日间精密度（分别≤2.3%和≤4.2%）和准确度（99.0-100.3%），与常规方法相比，可将 42 个样品的总 LC/ESI-MS/MS 运行时间减少 60%以上。因此，DAPPZ 同位素的衍生化对于提高胆汁酸的 LC/ESI-MS/MS 分析通量非常有效。
