A validated UHPLC-MS/MS method for quantification of total and free tedizolid concentrations in human plasma
A validated UHPLC-MS/MS method for quantification of total and free tedizolid concentrations in human plasma
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一种经过验证的 UHPLC-MS/MS 方法，用于定量人血浆中总泰地唑胺和游离泰地唑胺浓度
DOI：
10.1016/j.jpba.2022.114929
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发表时间：
2022
影响因子：
3.4
通讯作者：
Itoh Hiroki
中科院分区：
文献类型：
--
作者：
Tanaka Ryota;Kai Makoto;Shinohara Saori;Tatsuta Ryosuke;Itoh Hiroki
替地唑胺（TZD）是一种新型恶唑烷酮类抗生素。尽管TZD疗效与浓度-时间曲线下面积/最小抑制浓度相关，但尚无使用血浆游离TZD浓度进行药代动力学/药效学分析的报告。已经开发了几种方法来测量总TZD浓度，但没有测量游离TZD浓度。我们的目的是开发一种高通量的同时定量方法，使用超高效液相色谱串联质谱（UHPLC-MS/MS）的总和游离TZD浓度。采用平衡透析法分离游离组分。使用96孔HLB µElution板通过固相萃取进行预处理。使用C18柱进行分析物的色谱分离。在正离子模式下监测MS/MS转换。根据美国食品药品监督管理局（FDA）制定的生物分析方法验证指南进行全面验证。总TZD和游离TZD浓度分别在5-5000（r2= 0.9964）和1.5-1500（r2= 0.9990）ng/mL的宽范围内，该测定法显示出良好的线性。总浓度和游离浓度的批内准确度和精密度以及批间准确度和精密度符合FDA指南的标准。总浓度和游离浓度的回收率分别高于92.3%和85.3%。基质效应显示，TZD的总浓度和游离浓度在3个质控水平之间无显著差异，TZD的体外蛋白结合率为71.6%~ 76.9%，在校准范围内无浓度依赖性差异。接受TZD治疗的5例患者的总浓度和游离浓度均在校准曲线范围内，证明了新方法临床应用的可行性。总之，我们已经成功地开发了第一次的方法，同时定量的总和游离TZD浓度。
Tedizolid (TZD) is a novel oxazolidinone antibiotic. Although TZD efficacy correlates with area under the concentration-time curve/minimum inhibitory concentration, there is no report of pharmacokinetic/pharmacodynamic analysis using plasma free TZD concentrations. Several methods have been developed for measuring total TZD concentration, but not for free TZD concentration. We aimed to develop a high-throughput simultaneous quantification method for total and free TZD concentrations using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). The equilibrium dialysis method was used for separation of the free fraction. Pretreatment was conducted by solid-phase extraction using 96-well HLB µElution plate. Chromatographic separation of the analytes was conducted using a C18 column. MS/MS transitions were monitored in the positive ion mode. Full validation was performed in accordance with the bioanalytical method validation guidance prepared by the US Food and Drug Administration (FDA). The assay showed good linearity over wide ranges of 5–5000 (r2= 0.9964) and 1.5–1500 (r2= 0.9990) ng/mL for total and free TZD concentrations, respectively. Within-batch accuracy and precision as well as batch-to-batch accuracy and precision for total and free concentrations fulfilled the criteria of the FDA guidance. The recovery rates were higher than 92.3% and higher than 85.3% for total and free concentrations. Matrix effect showed no remarkable differences among three quality control levels for total and free concentrations.In vitroprotein binding rates of TZD ranged from 71.6% to 76.9%, indicating no concentration-dependent difference within the calibration ranges. The total and free concentrations in five patients who received TZD were within the ranges of the calibration curves, demonstrating the feasibility of clinical application of the novel method. In conclusion, we have succeeded to develop for the first time a method for simultaneous quantification of total and free TZD concentrations.