High-throughput simultaneous quantification of five azole anti-fungal agents and one active metabolite in human plasma using ultra-high-performance liquid chromatography coupled to tandem mass spectrometry
High-throughput simultaneous quantification of five azole anti-fungal agents and one active metabolite in human plasma using ultra-high-performance liquid chromatography coupled to tandem mass spectrometry
复制标题
使用超高效液相色谱-串联质谱法对人血浆中的五种唑类抗真菌剂和一种活性代谢物进行高通量同时定量
DOI：
10.1016/j.clinbiochem.2021.10.010
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发表时间：
2022
影响因子：
2.8
通讯作者：
Itoh Hiroki
中科院分区：
文献类型：
--
作者：
Tanaka Ryota;Shiraiwa Ken;Takano Kuniko;Ogata Masao;Honda Shuhei;Yoshida Natsumi;Okuhiro Kazuki;Yoshida Masaki;Narahara Kumiko;Kai Makoto;Tatsuta Ryosuke;Itoh Hiroki
目的：对于血液系统恶性肿瘤患者，预防深部真菌病常用的三唑类抗真菌药物有氟康唑(FLCZ)、伊曲康唑(ITCZ)、伏立康唑(VRCZ)、泊沙康唑(PSCZ)和异乌康唑(ISCZ)。由于这些唑类药物具有较大的药代动力学变异性，建议对某些唑类药物通过治疗药物监测进行剂量调整。本研究旨在建立并验证一种新的高效液相色谱-串联质谱法同时测定甲硝唑的血药浓度。设计与方法采用96孔MCX洗脱板的高通量固相萃取法作为前处理过程。线性范围分别为100-100000、20-20000、40-40000、20-20000、5-5000和50-50000 ng/m L，线性关系良好(校准器反算：相对误差≤为15%[≤20%])。所有6种药物的验证结果均符合美国食品和药物管理局关于批内和批间精密度和准确度的生物分析方法验证指南的标准。提取回收率较好，≥为74.9%，所有药物均未发现基质效应。用建立的方法对口服FLCZ、ITCZ、VRCZ或PSCZ的恶性血液病患者的谷值(1例PSCZ服药后10小时)药物浓度进行了定量。所有样品的测定均在标准曲线范围内，证明了该方法在临床应用的可行性。结论我们成功地建立了一种新的高通量的高效液相色谱-MS/MS法同时测定FLCZ、ITCZ、ITCZ-OH、VRCZ、PSCZ和ISCZ的血药浓度。
ObjectivesFor patients with hematological malignancy, triazole antifungal agents such as fluconazole (FLCZ), itraconazole (ITCZ), voriconazole (VRCZ), posaconazole (PSCZ) and isavuconazole (ISCZ) are often used for prophylaxis of deep mycosis. Since these azoles exhibit large pharmacokinetic variability, dose adjustment by therapeutic drug monitoring is recommended for some azoles. This study aimed to develop and validate a novel method for simultaneous determination of plasma concentrations of FLCZ, ITCZ, VRCZ, PSCZ, ISCZ and ITCZ-OH, an active metabolite of ITCZ, using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS).Design & methodsA high-throughput solid-phase extraction method using 96-well MCX µElution Plate was selected as the pretreatment procedure.ResultsThe calibration curves for FLCZ, ITCZ, ITCZ-OH, VRCZ, PSCZ and ISCZ showed good linearity (back-calculation of calibrators: relative error ≤ 15% [LLOQ: ≤ 20%]) over wide ranges of 100–100000, 20–20000, 40–40000, 20–20000, 5–5000 and 50–50000 ng/mL, respectively. The validation results of all six drugs fulfilled the criteria of the guidance for bioanalytical method validation of the US Food and Drug Administration for within-batch and batch-to-batch precision and accuracy. The extraction recovery rates were good at ≥ 74.9%, and almost no matrix effects were found for all the drugs. The trough (10 h post-dose in 1 patient on PSCZ) drug concentrations in patients with hematologic malignancy who received oral FLCZ, ITCZ, VRCZ or PSCZ were quantified using the method developed. The measurements for all samples were within the ranges of the calibration curves, demonstrating the feasibility of clinical application of the novel method.ConclusionsWe have succeeded in developing a novel high-throughput method using UHPLC-MS/MS for simultaneous quantification of plasma concentrations of FLCZ, ITCZ, ITCZ-OH, VRCZ, PSCZ and ISCZ.