湘葛一号根HPLC指纹图谱及5种异黄酮含量的同时测定  

HPLC fingerprint for roots of Hu′nan Pueraria thomsonii Benth and simultaneous determination of five isoflavones

作　　者：戴玲 叶惠煊[2,3] 邹亲朋 李芝[2] 刘向前[2] 

机构地区：[1]长沙博海生物科技有限公司,长沙410205 [2]湖南中医药大学药学院,长沙410208 [3]九芝堂股份有限公司,长沙410021

出　　处：《西北药学杂志》2016年第3期239-244,共6页Northwest Pharmaceutical Journal

基　　金：长沙市科技局科技计划重点项目(编号:K1403122-31)

摘　　要：目的采用高效液相色谱法建立湘葛一号根指纹图谱,并同时测定5种异黄酮的含量。方法色谱条件:采用AT·LICHROM C18色谱柱(250mm×4.6mm,5μm);以甲醇-水溶液为流动相进行梯度洗脱;流速为1.0mL·min^(-1);检测波长为250nm;柱温为30℃。对不同采收期的10批样品的图谱进行测定,运用中国药典委员会的"中药色谱指纹图谱相似度评价软件2004AB"进行评价,并对5种异黄酮成分进行含量测定。结果建立了湘葛一号根HPLC指纹图谱,各色谱峰分离度良好,确定了7个共有峰,并采用对照品指认了其中5个色谱峰。含量测定条件通过方法学验证,葛根素、大豆苷、染料木苷、大豆苷元、染料木素的线性范围分别为5.00~50.00,1.00~10.00,1.82~18.20,1.00~10.00和2.00~20.00μg·mL^(-1),线性关系良好(r>0.999),平均加样回收率为98.8%~101.8%,5种异黄酮含量测定结果可初步确定适宜采收期为12月份。结论该方法所建立的湘葛一号根指纹图谱特征性强、方法简单、结果可靠,结合主要有效成分含量测定可更好地用于其质量控制,对其鉴定也具有参考价值。Objective An HPLC fingerprint method for the roots of Hu′nan Pueraria thomsonii Benth in different harvest periods was developed,and quantities of five isoflavones were determined simultaneously.Methods The chromatography was performed on an AT·LICHROM C18column（250mm×4.6mm,5μm）with a mobile phase of methanol-water and gradient elution.The flow rate was 1.0mL·min^（-1）.The detection wavelength was 250 nm.The column temperature was 30 ℃.The chromatograms of 10 batches of samples in different harvest period was determined by HPLC.Results In the chromatographic fingerprint,7peaks were selected as the characteristic peaks to assess the similarities of different samples according to similarity evaluation for chromatographic fingerprint of traditional Chinese medicine（2004AB）.The chemical components contained in the roots of Hu′nan Pueraria thomsonii Benth were effectively separated.Five common fingerprint peaks were labeled from the seven common peaks.In quantitative analysis,the five components showed good regression（r0.999）,linear ranges of puerarin,daidzin,genistin,daidzein,genistein were 5.00-50.00,1.00-10.00,1.82-18.20,1.00-10.00 and 2.00-20.00μg·mL^-1,and their recoveries were in the range of 98.8%-101.8%,and the optimal harvest period was December.Conclusion The combination of chromatographic fingerprint and quantitative analysis can be readily used as a quality control method for Hu′nan Pueraria thomsonii Benth.

关 键 词：异黄酮 湘葛一号 高效液相色谱法 指纹图谱 

分 类 号：R284[医药卫生—中药学]