高效液相色谱法测定肾移植病人泼尼松龙、伏立康唑及其氮氧化物浓度

Determination of prednisolone,voriconazole and voriconazole N-oxide in kidney transplant patients by high-performance liquid chromatography

目的 建立快速、灵敏的高效液相色谱法测定病人血浆泼尼松龙、伏立康唑及其代谢物伏立康唑氮氧化物的浓度。方法色谱柱为安捷伦ZORBAX Eclipse XDB C_(18)(4.6 mm×250 mm,5μm);流动相为乙腈∶水(27∶73~60∶40)梯度洗脱;流速为1.0mL/min;波长为244 nm;柱温为30℃。收集2019年9月至2020年3月在安徽医科大学第一附属医院接受伏立康唑治疗的肾移植病人24例,检测24例肾移植病人血药浓度,男14例、女10例。结果 泼尼松龙、伏立康唑及其氮氧化物色谱峰分离良好。泼尼松龙在3.13~100.00μg/L范围内线性关系良好,r=0.993 1;伏立康唑在0.16~20.00 mg/L范围内线性关系良好,r=0.999 8;伏立康唑氮氧化物在0.16~10.00 mg/L范围内线性关系良好,r=0.997 3。三种化合物日内及日间RSD在1.93%~6.74%,绝对回收率在65.48%~86.68%。24例病人泼尼松龙浓度在7.75~72.75μg/L,伏立康唑浓度在0.17~5.44 mg/L,伏立康唑氮氧化物浓度在0.66~3.06 mg/L,个体差异较大。结论 该方法专属性强、灵敏度高,适合泼尼松龙、伏立康唑、伏立康唑氮氧化物血浆浓度的测定。

Objective plasma concentrations of prednisolone,voriconazole and voriconazole N-oxide in patients.Methods The chromatographic column was Agilent ZORBAX Eclipse XDB C_(18)(4.6 mm×250 mm,5 μm);the mobile phase was acetonitrile-water(27:73-60:40) gradient elution;the flow rate was 1.0 mL/min;the wavelength was 244 nm;and the column temperature was 30 ℃.A total of 24 kidney transplant patients who were hospitalized in the First Affiliated Hospital of Anhui Medical University and received voriconazole from September 2019 to March 2020 were enrolled,including 14 males and 10 females,and their blood drug concentrations were monitored.Results The chromatographic peaks of prednisolone,voriconazole and voriconazole N-oxide were well separated.The linear relationship of prednisolone was good in the range of 3.13-100.00 μg/L,r=0.993 1;the linear relationship of voriconazole was good in the range of 0.16-20.00 mg/L,r=0.999 8;the linear relationship of voriconazole N-oxide was good in the range of 0.16-10.00 mg/L,r=0.997 3.The intraday and interday RSDs of the three compounds ranged from 1.93% to 6.74%,and the absolute recoveries ranged from 65.48% to 86.68%.The concentration of prednisolone in 24 patients was 7.75-72.75 μg/L,the concentration of voriconazole was 0.17-5.44 mg/L,and the concentration of voriconazole N-oxide was 0.66-3.06 mg/L,with large individual differences.ConclusionThe method has strong specificity and high sensitivity and is suitable for the determination of plasma concentrations of prednisolone,voriconazole,and voriconazole N-oxide.