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

作　　者：党心 王仪 赵班[1] 毛永辉[1] DANG Xin;WANG Yi;ZHAO Ban;MAO Yong-hui(Department of Nephrology,Bejing Hospital,National Center of Gerontology,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Bejing 100730,China;School of Chemical and Environmental Engineering,China University of Mining and Technology,Beijing 100083,China)

机构地区：[1]北京医院,国家老年医学中心,中国医学科学院老年医学研究院肾内科,北京100730 [2]中国矿业大学化学与环境工程学院,北京100083

出　　处：《中国临床药理学杂志》2025年第5期717-721,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家科技重大专项“重大新药创制”子课题基金资助项目(2017ZX09304026);国家自然科学基金资助项目(81960752、81860727);中央高水平医院临床科研业务基金资助项目(121-2016008);中央高校基本科研业务费专项基金资助项目(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);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)。

摘　　要：目的建立一种超高效液相色谱串联高分辨率质谱法用于测定雷公藤多苷片治疗的糖尿病肾病患者血清中去甲泽拉木醛血药浓度的方法,并用于临床药效标志物的探索研究。方法以18O-去甲泽拉木醛为内标,血清样品经固相萃取处理后进样。色谱柱:BEH C18柱(2.1 mm×50.0 mm,1.7μm);流动相:0.1%甲酸水溶液-乙腈和0.1%甲酸水溶液;流速:0.3 mL·min^(-1);柱温:30℃;用电喷雾电离源,正离子多反应方式监测。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性。选择15例接受雷公藤多苷片治疗的糖尿病肾病患者,检测其血清去甲泽拉木醛的浓度。通过受试者工作特征(ROC)曲线和ROC曲线下面积(AUC)进行去甲泽拉木醛血药浓度与糖尿病肾病疗效的相关性分析。结果去甲泽拉木醛在1~200 ng·mL^(-1)线性关系良好,定量下限为1 ng·mL^(-1),日内、日间精密度相对标准差分别<10%和15%,绝对回收率达到89%以上,不受基质效应的影响,稳定性符合要求。去甲泽拉木醛血药浓度预测雷公藤多苷片疗效的AUC为0.66(95%置信区间为0.53~0.78),最佳截断值为61.73 ng·mL^(-1),敏感度为83.3%,特异度为46.7%(P<0.05)。结论本方法操作简单,灵敏度高,结果准确,适用于去甲泽拉木醛血药浓度的检测,可以用于雷公藤多苷片临床药效标志物的探索研究。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 C_(18)column(2.1 mm×50.0 mm,1.7μm)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

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 

分 类 号：R97[医药卫生—药品]