伏立康唑血浆药物浓度高效液相色谱法的建立及应用

Establishment and application of HPLC method for blood concentration of voriconazole

目的旨在开发一种测定伏立康唑血药浓度的方法,用于监测同时使用利福霉素类药物的患者体内伏立康唑血药浓度的变化。方法采用高效液相色谱法(HPLC)进行测定,色谱柱为Eclipse XDB-C18柱(4.6 mm×150 mm,5μm);流动相为甲醇/0.2%醋酸溶液(V/V=65:35);检测波长为256 nm;流速为1.0 mL/min;血浆样品经甲醇沉淀蛋白后,室温条件下进样分析。考察方法的专属性、线性、精密度与回收率等,并用建立的方法进行患者体内伏立康唑血药浓度的测定。结果标准曲线在0.204~10.2μg/mL浓度范围内线性关系良好(r=0.9999),伏立康唑保留时间为4.5 min,日内、日间精密度(RSD)与准确度(RE)均<10%,提取回收率为91.1%~95.1%。服用伏立康唑的21例结核患者中,17例患者联用利福霉素类药物,其中14例(82.4%)患者伏立康唑血药浓度低于有效范围下限;4例未联用利福霉素类药物的患者血药浓度在有效浓度范围内。服用伏立康唑的36例非结核患者中,仅5例(13.9%)患者伏立康唑血药浓度低于有效范围下限。结论本方法准确度高、稳定性好,简便快捷,适合于临床伏立康唑体内样本的高通量测定。

Objective The aim of this study was to develop a method for the determination of voriconazole concentration in plasma,which can be used to monitor the changes of voriconazole concentration in patients with tuberculosis after concurrent use of rifampicin.Methods The chromatographic column used was Eclipse XDB-C18 column(4.6 mm*150 mm,5 micron)by HPLC.The mobile phase was methanol/0.2%acetic acid solution(V/V=65:35).The detection wavelength was 256 nm.The flow rate was 1.0 mL/min.Plasma samples were injected and analyzed at room temperature after protein precipitation by methanol.The specificity,linearity,precision and recovery of the method were investigated and the plasma concentration of voriconazole was determined by the established method.Results The established method had a good linear relationship(r=0.9999)and a linear range of 0.204-10.2μg/mL,which covered the effective concentration range of voriconazole.The retention time of voriconazole was 4.5 min.The intra-day and inter-day precision(RSD)and accuracy(RE)were less than 10%,and the recovery was 91.1%-95.1%.During the sample determination,17 of 21 tuberculosis patients were treated with rifamycin.Among them,14 patients(82.4%)had blood concentration of voriconazole lower than the lower limit of effective range,and the blood concentrations of 4 patients without the use of rifamycin were within normal ranges.Among 36 non-tuberculosis patients,only 5(13.9%)patients had blood concentrations of voriconazole lower than the lower limit of effective range.Conclusion The established method was accurate,stable,simple and fast,and suitable for the high-throughput determination of voriconazole in vivo.