Institute for Instrumental Analytics and Bioanalysis, Mannheim University of Applied Sciences, Paul-Wittsack-Strasse 10, 68163, Mannheim, Germany.
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Iowa State University, 1809 South Riverside Drive, Ames, IA, 50011-1134, USA.
Anal Bioanal Chem. 2019 Aug;411(20):5187-5196. doi: 10.1007/s00216-019-01898-9. Epub 2019 May 23.
Paper spray high-resolution accurate mass spectrometry is a fast and versatile analysis method. This ambient ionization technique enables the quantitation of xenobiotics in complex biological matrices without chromatography or conventional sample extraction. The simplicity, rapidity, and affordability of the paper spray mass spectrometry (PS-MS) method make the technique especially attractive for clinical investigations where fast and affordable sample analysis is crucial. A new PS-MS method for the quantitation of voriconazole in equine tears was developed and validated. For a concentration range of 10 to 1000 ng/mL, good linearity (R > 0.99), inter- and intra-run precision (coefficient of variation (CV) max. 11.9%), accuracy (bias of the nominal concentration ± 13.9%), and selectivity (signal areas of the double blanks represent 0.13 ± 0.05% of the lower limit of quantitation (LLOQ) signal in equine tears) were observed. The quantitation of voriconazole was based on three product ions and calculated relative to the isotope-labeled internal standard, voriconazole-d, which had a final concentration of 250 ng/mL in the standards and samples. The matrix effect of the method showed an ionization suppression by reduction of the voriconazole response to 63.6%, 70.2%, and 81.9% for 30 ng/mL, 450 ng/mL, and 900 ng/mL in equine tears compared with voriconazole in solvent (methanol:water, 50:50, v:v). The method was used to analyze 126 study samples collected for a pharmacokinetic study investigating a novel approach for treatment of fungal keratitis in horses. Therefore, the integrity of the sample dilution (n = 6, CV 6.90%, and bias of nominal concentration + 8.40%) and the carryover effect (increase from 0.33 ± 0.21% to 1.33 ± 0.89% of the signal of the LLOQ) was further investigated. To our knowledge, this method is the first application of PS-MS for quantitation of drug concentrations in tears from any species.
纸喷雾高分辨率精确质量质谱是一种快速且多功能的分析方法。这种环境电离技术能够在无需色谱或常规样品提取的情况下，对复杂生物基质中的外源性物质进行定量分析。纸喷雾质谱（PS-MS）方法的简单、快速和经济实惠，使其特别适用于临床研究，在临床研究中，快速且经济实惠的样品分析至关重要。本文开发并验证了一种用于检测马眼泪中伏立康唑浓度的新 PS-MS 方法。在 10 至 1000ng/mL 的浓度范围内，观察到良好的线性（R>0.99）、重现性和日内精密度（CV 最大值为 11.9%）、准确度（标称浓度的偏差 ±13.9%）和选择性（空白信号的面积占马眼泪定量下限（LLOQ）信号的 0.13±0.05%）。伏立康唑的定量是基于三个产物离子，并相对于最终浓度为 250ng/mL 的同位素标记内标伏立康唑-d 进行计算。该方法的基质效应显示，与溶剂（甲醇：水，50:50，v:v）中的伏立康唑相比，马眼泪中 30ng/mL、450ng/mL 和 900ng/mL 的伏立康唑响应分别被抑制 63.6%、70.2%和 81.9%。该方法用于分析 126 个研究样本，这些样本是为一项研究新型治疗马属真菌角膜炎方法的药代动力学研究而采集的。因此，进一步研究了样品稀释的完整性（n=6，CV 为 6.90%，标称浓度的偏差为+8.40%）和交叉污染效应（从 LLOQ 信号的 0.33±0.21%增加到 1.33±0.89%）。据我们所知，这是 PS-MS 首次用于检测任何物种眼泪中药物浓度的应用。
