中国临床药理学杂志2025，Vol.41Issue(5)：717-721,5.DOI:10.13699/j.cnki.1001-6821.2025.05.023
用超高效液相色谱串联高分辨率质谱法测定去甲泽拉木醛的浓度
Determination of the concentration of demethylzeylasteral in serum by ultra-high performance liquid chromatography tandem high resolution mass spectrometry
摘要
Abstract
Objective To establish an ultra-high performance liquid chromatography tandem high resolution mass spectrometry(UHPLC-HRMS)method for the determination of serum demthylzeylasteral concentration in diabetic nephropathy patients,which treated by tripterygium wilfordii polyglycosides tablets(TGTs),and to apply this method in the exploratory study of clinical pharmacodynamic marker.Methods Serum samples and 18 O-demethylzeylasteral which was used as the internal standard were processed by solid phase extraction method and took the supernatant for the determination.Chromatographic separation of the analytes was accomplished using a BEH C18 column(2.1 mm × 50.0 mm,1.7 μm)under gradient elution,where the mobile phase consisted of a mixture of 0.1％formic acid aqueous solution and acetonitrile,as well as 0.1％formic acid aqueous solution.The flow rate was set at 0.3 mL·min-1 and the column temperature at 30 ℃.The mass spectrometer was equipped with an electrospray ionization source with positive multiple reaction monitoring mode.The specificity,standard curve and lower limit of quantification,precision and recovery rate,matrix effect and stability were investigated.This method was applied to determine the serum concentration of demethylzeylasteral from 15 patients with oral TGTs.Receiver operating characteristic(ROC)curve and area under the ROC curve(AUC)were utilized to explore the correlation between serum concentration of demethylzeylasteral and efficacy in treating diabetic nephropathy.Results The calibration curve of demethylezylasteral was linear over the range of 1-200 ng·mL-1,and the lower limit of quantification was 1 ng·mL-1.The precision and accuracy relative standard deviation of intra-day and inter-day was less than 10％and 15％,respectively.The average absolute recovery was more than 89％,no matrix effect has been affected and all stability tests met the acceptance criteria.The AUC of the efficacy of TGTs predicted by serum demethylzeylasteral quantitation was 0.66(95％confidence interval was 0.53-0.78),the cut-off value was 61.73 ng·mL-1,the sensitivity was 83.3％and the specificity was 46.7％(P＜0.05).Conclusion The UHPLC-HRMS method established in this study has the advantages of simple operation,high sensitivity,accurate and reliable results,which can be suitable for determining serum concentration of demethylzsylasteral and studying clinical drug efficacy markers of TGTs.关键词
去甲泽拉木醛/雷公藤多苷片/超高效液相色谱串联高分辨率质谱法/血药浓度/糖尿病肾病Key words
demethylzeylasteral/Tripterygium wilfordii polyglycoside tablet/ultra-high performance liquid chromatography tandem high resolution mass spectrometry/plasma drug concentration/diabetic nephropathy分类
医药卫生引用本文复制引用
党心,王仪,赵班,毛永辉..用超高效液相色谱串联高分辨率质谱法测定去甲泽拉木醛的浓度[J].中国临床药理学杂志,2025,41(5):717-721,5.基金项目
国家科技重大专项"重大新药创制"子课题基金资助项目(2017ZX09304026) （2017ZX09304026）
国家自然科学基金资助项目(81960752、81860727) （81960752、81860727）
中央高水平医院临床科研业务基金资助项目(121-2016008) （121-2016008）
中央高校基本科研业务费专项基金资助项目(2023ZKPYHH05) （2023ZKPYHH05）
中国医学科学院临床与转化医学研究专项基金资助项目(2023-I2M-C&T-B-116) （2023-I2M-C&T-B-116）
煤炭资源与安全开采国家重点实验室开放基金资助项目(SKLCRSM21KFA11) （SKLCRSM21KFA11）
