A highly sensitive method was developed to quantitate the antileishmanial agent paromomycin in human plasma, with a lower limit of quantification of 5 ng/mL. Separation was achieved using an isocratic ion-pair ultra-high performance liquid chromatographic (UPLC) method with a minimal concentration of heptafluorobutyric acid, which was coupled through an electrospray ionization interface to a triple quadrupole - linear ion trap mass spectrometer for detection. The method was validated over a linear calibration range of 5 to 1000 ng/mL (r2≥0.997) with inter-assay accuracies and precisions within the internationally accepted criteria. Volumes of 50 μL of human K2EDTA plasma were processed by using a simple protein precipitation method with 40 μL 20 % trichloroacetic acid. A good performance was shown in terms of recovery (100 %), matrix effect (C.V. ≤ 12.0 %) and carry-over (≤17.5 % of the lower limit of quantitation). Paromomycin spiked to human plasma samples was stable for at least 24 h at room temperature, 6 h at 35 °C, and 104 days at -20 °C. Paromomycin adsorbs to glass containers at low concentrations, and therefore acidic conditions were used throughout the assay, in combination with polypropylene tubes and autosampler vials. The assay was successfully applied in a pharmacokinetic study in visceral leishmaniasis patients from Eastern Africa.

中文翻译：

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高灵敏度的UPLC-MS / MS方法，用于定量人血浆中巴龙霉素。

开发了一种高灵敏度的方法来定量人血浆中的抗疟原虫药帕罗霉素，其定量下限为5 ng / mL。使用等浓度离子对超高效液相色谱（UPLC）方法和最小浓度的七氟丁酸进行分离，该方法通过电喷雾电离接口与三重四极杆-线性离子阱质谱仪耦合进行检测。该方法在5至1000 ng / mL的线性校准范围内（r2≥0.997）进行了验证，其批间准确性和精密度均在国际公认的标准范围内。通过使用简单的蛋白质沉淀方法，用40μL20％三氯乙酸处理50μL人K2EDTA血浆。在回收率（100％），基质效应（CV）方面显示出良好的性能 ≤12.0％）和残留（≤定量下限的17.5％）。掺入人血浆样品的巴龙霉素在室温下至少稳定24小时，在35°C稳定6小时，在-20°C稳定104天。巴龙霉素以低浓度吸附到玻璃容器中，因此在整个分析过程中使用酸性条件，并与聚丙烯试管和自动进样器小瓶结合使用。该测定法已成功用于东非内脏利什曼病患者的药代动力学研究。与聚丙烯管和自动进样器样品瓶结合使用。该测定法已成功用于东非内脏利什曼病患者的药代动力学研究。与聚丙烯管和自动进样器样品瓶结合使用。该测定法已成功用于东非内脏利什曼病患者的药代动力学研究。