目的:分别建立超滤法和溶剂萃取法处理样品,结合高效液相色谱(HPLC)法测定人血浆中伏立康唑游离型药物浓度Cu和总血药浓度Ct,并计算蛋白结合率。方法:游离型药物浓度采用Millipore Centrifree超滤装置前处理,以外标法测定;总血药浓度测定采用内标法,经蛋白沉淀、溶剂萃取、复溶后测定。结果:游离型药物浓度和总血药浓度的线性范围分别为0.20~12.64μg·mL^-1和0.02~10.24μg·mL^-1;游离型药物浓度和总血药浓度的准确度和精密度均在±15%范围内,ICU患者的伏立康唑血浆蛋白结合率为42.4%。结论:本方法前处理方法便捷,选择性强,准确度高,可用于人血浆中伏立康唑蛋白结合率的测定。
OBJECTIVE To establish an ultrafiltration method and solvent extraction method to process samples,combine high performance liquid chromatography(HPLC)method to measure the unbound(Cu)and total(Ct)voriconazole concentrations simultaneously,and calculate plasma protein binding(PPB).METHODS Cuof voriconazole was treated with the Millipore Centrifree by external standard method.Ctof voriconazole was determined by using an internal standard method after protein precipitation,solvent extraction and redissolution.RESULTS RESULTS The standard curves of unbound and total concentrations were liner in the range of 0.20-12.64μg·mL^-1 and 0.02-10.24μg·mL^-1,respectively.The intra-and inter-day precision and accuracy were with ±15% of reference value,the median PPB was 42.4%in ICU sample.CONCLUSION The described method offers a convenient and sensitive analysis for PPB of voriconazole in human plasma.