Koster Remco A, Niemeijer Pascal, Veenhof Herman, Hateren Kai van, Alffenaar Jan-Willem C, Touw Daan J
Department of Clinical Pharmacy & Pharmacology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
Bioanalytical Laboratory, PRA Health Sciences, Amerikaweg 18, 9407 TK, Assen, The Netherlands.
Bioanalysis. 2019 Mar;11(6):495-508. doi: 10.4155/bio-2018-0312. Epub 2019 Mar 20.
The aim of this study was to develop and validate a LC-MS/MS assay for tacrolimus, sirolimus, everolimus, cyclosporin A and mycophenolic acid using volumetric absorptive microsampling tips as a sampling device and to investigate the effect on the recoveries of the analyte concentration in combination with the hematocrit (HT), which included temsirolimus (a structural analog). The maximum observed overall bias was 9.6% for the sirolimus LLOQ, while the maximum overall coefficient of variation was 8.3% for the everolimus LLOQ. All five immunosuppressants demonstrated to be stable in the volumetic absorbtive microsampling tips for at least 14 days at 25°C. Biases caused by HT effects were within 15% for all immunosuppressants between HT levels of 0.20 and 0.60 l/l, except for cyclosporin A, which was valid between 0.27 and 0.60 l/l. Reduced recoveries were observed at high analyte concentrations in combination with low HT values for sirolimus, everolimus and temsirolimus. A robust extraction and analysis method in volumetric absorptive microsampling tips was developed and fully validated. HT- and concentration-related recovery effects were observed but were within requirements of the purpose of the analytical method.
本研究的目的是开发并验证一种液相色谱-串联质谱法（LC-MS/MS），用于测定他克莫司、西罗莫司、依维莫司、环孢素A和霉酚酸，使用体积吸收微采样尖端作为采样装置，并研究血细胞比容（HT）对分析物浓度回收率的影响，其中包括替西罗莫司（一种结构类似物）。西罗莫司定量下限（LLOQ）的最大观察到的总偏差为9.6%，而依维莫司LLOQ的最大总变异系数为8.3%。所有五种免疫抑制剂在体积吸收微采样尖端中于25°C下至少14天内均表现出稳定性。除环孢素A在0.27至0.60 l/l之间有效外，所有免疫抑制剂在HT水平为0.20至0.60 l/l之间由HT效应引起的偏差均在15%以内。对于西罗莫司、依维莫司和替西罗莫司，在高分析物浓度与低HT值相结合时观察到回收率降低。开发并充分验证了一种在体积吸收微采样尖端中稳健的提取和分析方法。观察到与HT和浓度相关的回收率效应，但在分析方法目的的要求范围内。
