Zimbelmann Franziska, Groll Andreas H, Hempel Georg
Department of Pharmaceutical and Medical Chemistry-Clinical Pharmacy, University of Münster, 48149 Münster, Germany.
Infectious Disease Research Program, Center for Bone Marrow Transplantation and Department of Paediatric Haematology/Oncology, University Children's Hospital Münster, 48149 Münster, Germany.
Pharmaceutics. 2025 May 1;17(5):592. doi: 10.3390/pharmaceutics17050592.
: A simple method for quantifying fluconazole in small blood volumes has been developed using volumetric absorptive microsampling (VAMS) technology and isocratic high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection. : For sample collection, Mitra devices are used to keep the sample volume at 10 µL. For the quantitative determination of fluconazole, the Mitra samples are extracted using acetonitrile as the extraction agent, containing 2-(4-chlorophenyl)-1,3-bis(1,2,4-triazol-1-yl)propan-2-ol as the internal standard. A Synergi 4 μm Polar-RP 80 Å (150 × 2 mm) column forms the stationary phase, and a mixture of acetonitrile and phosphate buffer is the mobile phase. The UV detection is set at a wavelength of 210 nm. The therapeutic concentration range of 5 to 160 mg/L is covered, and the linear equation with 1/x weighting is used to determine unknown samples. This method has been validated according to the current EMA and FDA guidelines for bioanalytical methods. : The validation data obtained after analysing whole blood samples (EDTA) showed within- and between-run accuracy between 94.4% and 115% and precision between 0.4% and 9.4%, respectively. A lower limit of quantification (LLOQ) of 5 mg/L was sufficient for therapeutic drug monitoring in paediatric patients receiving fluconazole as antifungal prophylaxis after haematopoietic cell transplantation. : So far, 211 samples from 49 patients were successfully analysed, and concentrations between 5.84 mg/L and 107 mg/L were determined for whole blood Mitra samples. To our knowledge, this is the first application of VAMS technology using simple and cheap HPLC-UV quantification.
已开发出一种使用体积吸收微采样（VAMS）技术和带有紫外（UV）检测的等度高效液相色谱（HPLC）来定量测定少量血样中氟康唑的简单方法。对于样本采集，使用Mitra装置将样本体积保持在10μL。为了定量测定氟康唑，使用乙腈作为萃取剂对Mitra样本进行萃取，乙腈中含有2-（4-氯苯基）-1,3-双（1,2,4-三唑-1-基）丙-2-醇作为内标。一根Synergi 4μm Polar-RP 80Å（150×2mm）色谱柱构成固定相，乙腈和磷酸盐缓冲液的混合物作为流动相。UV检测设置在210nm波长处。该方法涵盖了5至160mg/L的治疗浓度范围，并使用带1/x加权的线性方程来测定未知样本。此方法已根据当前欧洲药品管理局（EMA）和美国食品药品监督管理局（FDA）的生物分析方法指南进行了验证。分析全血样本（乙二胺四乙酸抗凝）后获得的验证数据显示，批内和批间准确度分别在94.4%至115%之间，精密度分别在0.4%至9.4%之间。5mg/L的定量下限（LLOQ）足以对接受氟康唑作为造血细胞移植后抗真菌预防的儿科患者进行治疗药物监测。到目前为止，已成功分析了来自49名患者的211个样本，测定的全血Mitra样本浓度在5.84mg/L至107mg/L之间。据我们所知，这是首次使用简单且廉价的HPLC-UV定量方法应用VAMS技术。
