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作 者:刘廷[1] 狄留庆[1] 李娟[1] 康安[1] 李俊松[1] 赵晓莉[1] 单进军[1]
机构地区:[1]南京中医药大学药学院,江苏省中药高效给药系统工程技术研究中心,南京210023
出 处:《中国实验方剂学杂志》2016年第18期82-87,共6页Chinese Journal of Experimental Traditional Medical Formulae
基 金:江苏省“333高层次人才培养工程”项目(BRA2013201);江苏高校优势学科建设工程项目(ysxk-2010)
摘 要:目的:建立同时测定大鼠灌服双黄连制剂后血浆中酚酸类(新绿原酸、绿原酸及隐绿原酸)与黄酮类(黄芩苷、千层纸素A-7-O-β-D-葡萄糖苷、汉黄芩苷、黄芩素、汉黄芩素及千层纸素A)成分的超高效液相色谱-串联质谱法(UPLC-MS/MS),并研究其药代动力学。方法:血浆样品采用乙酸乙酯-正丁醇(1∶1)萃取,流动相0.4%甲酸水溶液-[乙腈-甲醇(4∶1)混合液]梯度洗脱,流速0.4 m L·min-1,采用电喷雾离子化,多反应监测离子扫描模式,选用响应较高的正离子模式。结果:建立的同时测定血浆中9种成分的UPLC-MS/MS线性良好(R2〉0.992),精密度RSD均〈14.2%,准确度85.49%~114.76%,复方中新绿原酸、绿原酸、隐绿原酸、黄芩苷、千层纸素A-7-O-β-D-葡萄糖苷、汉黄芩苷、黄芩素的药时曲线下面积AUC0-t与AUC0-∞与单味药提取物相比显著增加,而酚酸和黄酮类成分的消除速率较单味药提取液缓慢。结论:建立的UPLC-MS/MS特异、快速、准确及灵敏,可用于双黄连制剂中9种主要活性成分在大鼠体内的药代动力学研究。Objective: A rapid and sensitive ultra performance liquid chromatography tandem mass spectrometry method( UPLC-MS / MS) was developed and validated for simultaneously determination of phenolic acids and flavonoids in rat plasma after oral administration of Shuanghuanglian preparation. Method: Plasma samples were pretreated by n-butanol-ethyl acetate( 1 ∶ 1). The separation was preformed on an Waters Acquity HSS T3 C18column( 2. 1 mm × 100 mm,1. 7 μm) and the flow rate was set at 0. 4 m L·min- 1with mobile phase of acetonitrile-methanol( 4 ∶ 1)-0. 4% formic acid for gradient elution. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring( MRM) via electrospray ionization( ESI)source with positive mode. Result: All calibration curves had good linearity( R2 0. 992). RSDs of precisionand accuracy were all satisfied. After oral administration of Shuanghuanglian preparation,a remarkable increase in values of AUC0-t and AUC0-∞of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,baicalin,oroxylin A-7-O-β-D-glucuronide,wogonoside and baicalein was observed by compared with Lonicerae Japonicae Flos extract group or Scutellariae Radix extract group; but the elimination rate of phenolic acids and flavonoids in Shuanghuanglian preparation was slower than the single herb extract. Conclusion: The developed method is successfully applied to pharmacokinetic study of nine active ingredients in Shuanghuanglian preparation,this study can provide experimental evidence for elucidating rationality of compatibility of this preparation.
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