葛根芩连汤中14种主要成分在大鼠体内的药动学特征研究  

Pharmacokinetic characteristics of fourteen main components of Gegen Qinlian Decoction in rats

作　　者：王新雨[1,2,3] 晋臻 凌霄 刘昌顺[1,2,3] 谭晓梅 WANG Xin-yu;JIN Zhen;LING Xiao;LIU Chang-shun;TAN Xiao-mei(School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China;Guangdong Provineal Key Laboratory of Chinese Medicine Pharmaceutics,Guangzhou 510515,China;Guangdong Provincal Engineering Laboratory of Chinese Medicine Preparation Technology,Guangzhou 510515,China)

机构地区：[1]南方医科大学中医药学院,广东广州510515 [2]广东省中药制剂重点实验室,广东广州510515 [3]广东省中药制剂技术工程实验室,广东广州510515

出　　处：《中国中药杂志》2018年第23期4724-4734,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81374049)

摘　　要：该研究采用超高效液相色谱-串联质谱(UPLC-MS/MS)方法,研究大鼠单次灌胃葛根芩连汤(15 mL·kg^-1)后血浆中葛根素、3′-羟基葛根素、大豆苷、大豆苷元、染料木苷、染料木素、黄芩苷、黄芩素、汉黄芩苷、汉黄芩素、小檗碱、药根碱、巴马汀、甘草苷14种主要成分的药动学特征。采用Zorbax SB-18色谱柱以乙腈(A)-0.1%甲酸水溶液(B)为流动相对分析物和内标(柚皮苷和泼尼松龙)进行梯度洗脱:0~2.5 min,15%~30%A;2.5~3.5 min,30%~35%A;3.5~5.0 min,35%~40%A;5.0~9.0 min,40%~60%A;9.0~11.0 min,60%~15%A。流速为0.4 mL·min^-1。采用电喷雾电离(ESI)源以多反应监测(MRM)模式进行正负离子检测。结果14种成分的线性相关系数大于0.99,血浆样品反复冻融3次、-20℃放置7 d、室温放置12 h以及血浆样品处理后放置24 h的日内、日间变异系数(RSD)小于10%,精密度和准确度等均符合生物样品分析的要求。表明该方法灵敏、准确,适用于葛根芩连汤中14种成分的药动学研究。采用DAS 3.2.2软件的非房室模型计算药动学参数,结果异黄酮类成分和黄酮类成分的MRT0-t在7.5~11.8,T1/2z主要在11.0~29.7 h,黄酮类AUC0-t大于异黄酮类。而生物碱类成分的MRT0-t在4.3~7.2 h,T1/2z在1.0~5.0 h,AUC0-t小于黄酮类和异黄酮类成分。结果提示黄酮及异黄酮类成分体内血药浓度高、作用时间长,有利于抗炎解热作用的发挥;生物碱类成分体内血药浓度低,作用时间短,可能在肠道内发挥其抑菌作用。A specific and selective UPLC-MS/MS method was developed and validated for the simultaneous determination of isoflavonoids(3′-hydroxy puerarin, puerarin, daidzin, daidzein, genistin, genistein), flavonoids(baicalin, baicalein, wogonoside, wogonin, liquiritin)and alkaloids(berberine, jatrorrhizine, palmatine)(14 bioactive compounds) of Gegen Qinlian Decoction(GQD)in plasma. The pharmacokinetics characteristics of 14 bioactive compounds were study after oral administration of GQD at a single dose to rats. Prednisolone was used as the internal standard of liquiritin, and naringin was used as the internal standard of the other thirteen analytes. After the plasma samples were processed by precipitation protein method, the constituents and internal standards were gradient eluted by using a Zorbax SB-18 column with a mobile phase of acetonitrile(A) and 0.1% formic acid aqueous solution(B) using a gradient elution of 0-2.5 min,15%-30% A;2.5-3.5 min,30%-35% A;3.5-5.0 min,35%-40% A;5.0-9.0 min,40%-60% A;9.0-11.0 min,60%-15% A,and the flow rate was 0.4 mL·min^-1. The auto sampler was conditioned at 25 ℃ and the sample injection volume was 5 μL. A mass spectrometry was applied with electrospray ionization(ESI) ion source in the positive and negative ion multiple reaction monitoring(MRM) mode. All pharmacokinetic parameters were processed by non-compartmental analysis with DAS 3.2.2 software. The results showed that the linear correlation coefficient of the 14 components were all greater than 0.99, indicating that the method had good linearity in their respective concentration ranges. Post-preparative stability(25 ℃, 24 h), short-term stability(25 ℃, 12 h), long-term stability(-20 ℃, 7 d), and freeze and thaw stability(3-cycles) of the fourteen constituents were examined to evaluate the stability of methodology. The results of the inner and inter-day relative standard deviations were both less than 10%, indicating legitimate precise and accuracy to the requirement of biological sample analysis. The assay method is

关 键 词：葛根芩连汤 超高效液相色谱-串联质谱 药代动力学 

分 类 号：R285.5[医药卫生—中药学]