中国中药杂志

2016, v.41(24) 4579-4586

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6种黄连饮片中6种生物碱的RP-HPLC含量测定及与“治消渴”药效学的谱-效关系分析
Determination of six alkaloids in six types of Coptidis Rhizoma pieces by RP-HPLC and spectrum-effect relationships with anti-diabetes pharmacodynamics data

赖先荣;周邦华;杜明胜;郑海杰;耿志鹏;李佳川;孟宪丽;张艺;张静;
LAI Xian-rong;ZHOU Bang-hua;DU Ming-sheng;ZHENG Hai-jie;GENG Zhi-peng;LI Jia-chuan;MENG Xian-li;ZHANG Yi;ZHANG Jing;Chengdu University of Traditional Chinese Medicine;

摘要(Abstract):

建立生品、姜炙、醋炙、酒蒸、酒炙、萸炙6种黄连饮片中6种生物碱(盐酸药根碱、盐酸非洲防己碱、盐酸表小檗碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱)的含量测定方法,并结合药效学结果探索其与药效学研究结果及中医疗效之间的关系。采用Welch XtimateTMC18(4.6 mm×250 mm,5μm)色谱柱,以0.1%三乙胺溶液(用碳酸氢铵和氨水调节p H 10)为流动相A,乙腈为流动相B,进行梯度洗脱(0~15 min,10%~25%B;15~25 min,25%~30%B;25~40 min,30%~45%B),流速1.0 m L·min-1,柱温30℃,检测波长270 nm。6种生物碱分别在0.85~16.96 mg·L-1(r=0.999 7),1.25~24.96 mg·L-1(r=0.999 9),2.05~40.96 mg·L-1(r=0.999 9),3.65~72.96 mg·L-1(r=0.999 9),2.88~57.06 mg·L-1(r=0.999 8),13.25~264.96 mg·L-1(r=0.999 6)呈现良好的线性关系,平均加样回收率(n=9)分别为102.4%(RSD 1.2%),101.8%(RSD 1.3%),100.3%(RSD 1.8%),100.7%(RSD 1.8%),101.2%(RSD 1.5%),97.90%(RSD 2.0%)。测定6种黄连饮片共36个批次的6种生物碱的平均质量分数分别为3.55,4.49,9.12,19.17,15.69,62.56 mg·g-1。该文建立含量测定方法准确性高、重复性好,可用于6种黄连饮片中6种生物碱的含量测定。采用主成分分析、分层聚类分析对含量测定及前期药效学研究结果进行数据分析,结果表明酒蒸、酒炙、萸炙3种黄连饮片与生品黄连饮片存在明显差异,酒蒸饮片与生品饮片具有最大的差异,其差异主要与血清甘油三酯(TG)、空腹血糖水平(FBG)有关,另外,盐酸非洲防己碱是6种生物碱成分中影响最大的生物碱成分。酒蒸、酒炙、萸炙3种黄连饮片用于中医临床"治消渴"(防治糖尿病)更有优势,特别是在治疗糖尿病高脂血症方面。
To establish a method for determining the contents of six alkaloids( jatrorrhizine hydrochloride,columbamine hydrochloride,epiberberine hydrochloride,coptisine hydrochloride,palmatine hydrochloride,berberine hydrochloride) in six types of Coptidis Rhizoma pieces( crude pieces,ginger juice stir-fried pieces,vinegar stir-fried pieces,wine steamed pieces,wine stir-fried pieces,evodiae juice stir-fried pieces) by RP-HPLC,and explore the relationship with the curative effect of traditional Chinese medicine( TCM) and pharmacodynamics results. The chromatographic column was Welch XtimateTMC18( 4. 6 mm × 250 mm,5 μm),with0. 1% triethylamine solution( adjust p H at 10 with ammonium bicarbonate and ammonia) as mobile phase A and acetonitrile as mobile phase B for gradient elution( 0-15 min,10%-25% B; 15-25 min,25%-30% B; 25-40 min,30%-45% B) at a rate of 1. 0m L·min-1. The column temperature was set at 30 ℃,and the wavelength was set at 270 nm. The six alkaloids showed a good linear relationship within the range of 0. 85-16. 96 mg· L-1( r = 0. 999 7),1. 25-24. 96 mg·L-1( r = 0. 999 9),2. 05-40. 96 mg·L-1( r = 0. 999 9),3. 65-72. 96 mg·L-1( r = 0. 999 9),2. 88-57. 60 mg·L-1( r = 0. 999 8),and 13. 25-264. 96 mg·L-1( r = 0. 9996) respectively. The average recoveries( n = 9) of the six alkaloids were 102. 4%( RSD 1. 2%),101. 8%( RSD 1. 3%),100. 3%( RSD 1. 8%),100. 7%( RSD 1. 8%),101. 2%( RSD 1. 5%) and 97. 90%( RSD 2. 0%) respectively,and their average contents were 3. 55,4. 49,9. 12,19. 17,15. 69,62. 56 mg · g-1,respectively. This determination method was accurate and repeatable,which could be used for the content determination in six types of Coptidis Rhizoma pieces. Data analysis on contents determination and preliminary pharmacodynamics results was conducted by using principal component analysis( PCA) and hierarchical clustering analysis( HCA). The analysis results showed that three types of Coptidis Rhizoma pieces( wine steamed pieces,wine stir-fried pieces,and evodiae juice stir-fried pieces) had significant differences with crude pieces,and the wine steamed Coptidis Rhizoma pieces showed most difference with crude pieces especially,mainly related to triglyceride( TG) and fasting blood glucose levels( FBG) in serum. In addition,columbamine hydrochloride was most affected among the six alkaloids. Those three types of Coptidis Rhizoma pieces( wine steamed pieces,wine stir-fried pieces,and evodiae juice stir-fried pieces),had more advantages for " anti-diabetes" in TCM clinical application,especially in the treatment of diabetic hyperlipidemia.

关键词(KeyWords): 黄连饮片;HPLC;盐酸药根碱;盐酸非洲防己碱;盐酸表小檗碱;盐酸黄连碱;盐酸巴马汀;盐酸小檗碱;含量测定;药效学;主成分分析;分层聚类分析
Coptidis Rhizoma pieces;HPLC;jatrorrhizine hydrochloride;columbamine hydrochloride;epiberberine hydrochloride;coptisine hydrochloride;palmatine hydrochloride;berberine hydrochloride;content determination;pharmacodynamics data;principal components analysis(PCA);hierarchical clustering analysis(HCA)

Abstract:

Keywords:

基金项目(Foundation): 国家“十一五”科技支撑计划项目(2007BAI40B05,2007BAI40B03)

作者(Author): 赖先荣;周邦华;杜明胜;郑海杰;耿志鹏;李佳川;孟宪丽;张艺;张静;
LAI Xian-rong;ZHOU Bang-hua;DU Ming-sheng;ZHENG Hai-jie;GENG Zhi-peng;LI Jia-chuan;MENG Xian-li;ZHANG Yi;ZHANG Jing;Chengdu University of Traditional Chinese Medicine;

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