基于标准加入法和多水平的药代动力学综合分析疏血通注射液中不稳定的次黄嘌呤(潜在的质量标志物)  

The integrated analysis strategy of unstable hypoxanthine,a potential quality marker in Shuxuetong injection based on standard addition method and multi-level pharmacokinetics by LC-MS/MS

作　　者：邢彦超 王晓明[1] 王献瑞 赵西子 郭亚卿[1] 黄宇虹[3] Tekleab Teka 韩立峰[1,2] 潘桂湘 Yanchao Xing;Xiaoming Wang;Xianrui Wang;Xizi Zhao;Yaqing Guo;Yuhong Huang;Tekleab Teka;Lifeng Han;Guixiang Pan(Tianjin University of Traditional Chinese Medicine,Tianjin,China;State Key Laboratory of Component-based Chinese Medicine,Tianjin Key Laboratory of TCM Chemistry and Analysis,Tianjin,China;Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Hebei District,Tianjin,China)

机构地区：[1]天津中医药大学,天津 [2]天津中医药大学中医药研究院组分中药国家重点实验室,天津 [3]天津中医药大学第二附属医院,天津

出　　处：《Acupuncture and Herbal Medicine》2023年第2期116-125,共10页针灸和草药（英文）

基　　金：supported by the National Natural Science Foundation of China(81303182,81173523);the National Major Scientific and Technological Special Project for“Significant New Drugs Development”(2018ZX09734-002).

摘　　要：[目的]作为一种中药注射制剂,舒血通注射液用于治疗缺血性脑卒中。次黄嘌呤是其潜在的质量标志物之一。本研究的目的是通过对次黄嘌呤的定量和药代动力学行为分析为疏血通注射液的质量控制奠定基础。[方法]应用液相色谱-串联质谱(LC-MS/MS)首次建立了基于标准加入法的疏血通注射液中次黄嘌呤的定量方法。另一方面,也成功建立了基于LC-MS/MS的大鼠给药后血浆样品中次黄嘌呤的测定方法。[结果]对于相同批次的疏血通注射液,采用常规的液质联用技术测出的次黄嘌呤的浓度大于LC-MS/MS联合标准加入法测出的次黄嘌呤的浓度。单次静脉注射疏血通注射液的低、中、高剂量比为1:2:4,AUC0-t分别为(848.34±324.53)μg·h/L、(1483.94±497.74)μg·h/L,(3074.84±910.29)μg·h/L,剂量呈良好的线性依赖关系。[结论]标准加入法的引入可校正目标化合物的浓度,进而消除内源性物质的影响。别嘌呤出可抑制次黄嘌呤的转化以保证药代动力学研究中检测的准确性。通过空白血浆前处理获得的“空白生物基质”成功区分了内源性和药源性的次黄嘌呤。静脉注射次黄嘌呤血药浓度与给药剂量呈良好的线性关系。同样,多次给药中剂量组也没有药物蓄积,这与单次给药中剂量组的药代动力学特征是相似的。Objective:As an injection made from traditional Chinese medicine,Shuxuetong(SXT)injection is used for the treatment of ischemic stroke.Hypoxanthine is regarded as one of its potential quality markers.The purpose of this study is to lay the foundation for the quality control of SXT injection by the analysis of the quantitation and pharmacokinetic behavior of hypoxanthine.Methods:A quantitative method of hypoxanthine in SXT injection based on standard addition method by liquid chromatography-tandem mass spectrometry(LC-MS/MS)was established for the first time.On the other hand,a determination method of hypoxanthine in rat plasma samples after administration of SXT was also successfully established based on LC-MS/MS.Results:It was found that the content of hypoxanthine was higher using conventional liquid-mass spectrometry technology compared to the application of LC-MS/MS combined standard addition method in the same batch of SXT injection.The ratio of low,medium and high doses of intravenous SXT were 1:2:4,and the AUC0-t was(848.34±324.53)μg·h/L,(1483.94±497.74)μg·h/L,and(3074.84±910.29)μg·h/L,respectively.AUC_(0-t)shows a good linear dose-dependent relationship.Conclusions:The influences of endogenous substances tend to be eliminated by calibrating the concentration level of the target compound by the introduction of the standard addition method.The added allopurinol could inhibit the conversion of the target compound,and ensure the accuracy of the detection during the pharmacokinetic studies.“Blank biological matrix”obtained from the pretreatment of blank plasma successfully distinguished endogenous and drug-derived hypoxanthine.There is a good linear relationship between the blood concentration of intravenous hypoxanthine and the dosage of administration.Similarly,there was no drug accumulation in the multiple medium-dosage group,which is similar to the pharmacokinetic characteristics of the single medium-dosage group.

关 键 词：次黄嘌呤 LC-MS/MS 药代动力学 稳定性 标准加入法 疏血通注射液 

分 类 号：R285.5[医药卫生—中药学]