对照品替代法同时测定中药和保健食品中6种大豆异黄酮类成分的含量___

摘要 目的：尝试对照品替代法在高效液相色谱法含量测定中的应用，即建立以1种成分为对照同时测定多种目标成分的含量测定方法。方法：采用HPLC-DAD-MS联用技术，通过定性和定量手段分别求得各被测成分相对于特定对照品的计算因子，从而实现以染料木素为对照，同时测定样品中大豆苷、黄豆黄苷、染料木苷、大豆苷元、黄豆黄素和染料木素6种成分的含量。色谱条件：采用Agilent Zorbax SB-C₁₈(4.6 mm×150 mm,5 μm)柱,柱温25 ℃；流动相为0.2%甲酸-乙腈，梯度洗脱；流速为1 mL·min^-1(DAD检测器)及0.2 mL·min^-1(质谱检测器)。质谱条件：ESI正离子模式检测，毛细管电压3.0 kV，锥孔电压40 V，源温度120 ℃，去溶剂温度150 ℃，去溶剂气(N₂)流速300 L·h^-1，离子扫描范围m/z 100~700；选定m/z 255,271,285,417，433,447进行选择性离子扫描(SIM)。结果：以对照品替代法测得的结果与常规的含量测定方法测得的结果基本一致。结论：本方法经济、实用、快速、高效，可用于含大豆异黄酮类中药和保健食品中以上6种成分的含量测定和产品质量控制，同时可解决因对照品不足而给含量测定带来的困难，大大降低了分析检测的成本。

Abstract Objective:To develop a method for simultaneous determination of 6 isoflavones in traditional Chinese medicine and functional foods with one chemical reference substance,which is named as replacement method.Methods:A series of calculator correct factors were obtained by HPLC-DAD-MS,and the contents of 6 isoflavones were quantified with genistein as standard reference.HPLC-DAD-MS condition,column:Agilent Zorbax SB-C₁₈,4.6 mm×150 mm,5 μm;column temperature:25 ℃;mobile phase:0.2% formic acid-acetonitrile with a linear gradient elution;flow rate:1.0 mL·min^-1(DAD),0.2 mL·min^-1(MS).MS detection mode:electrospray positive ion;capillary voltage:3.0 kV;cone voltage:40 V;temperature of ESI probe:120 ℃;desolvation temperature at 150 ℃;desolvation gas flow:300 L·h^-1;full scan:100-700 amu;selected ion monitoring(SIM,m/z):255,271,285,417,433,and 447.Results:The quantitative results with new method were almost same to the results with conventional HPLC method.Conclusion:A economic,sensitive,simple and practicable HPLC-DAD-MS method was established for the determination of 6 isoflavones in traditional Chinese medicine and functional foods with one chemical reference substance.