HPLC法分离18α-、18β-甘草酸并用于甘草酸类产品的质量控制  

HPLC determination of 18α-and 18 β-glycyrrhizin for quality control of glycyrrhizin products

作　　者：吕昭云 吕亚光 车宝泉[2] 张世勇 张猛 潘显道 

机构地区：[1]北京协和药厂,北京102600 [2]北京市药品检验所,北京100035

出　　处：《药物分析杂志》2010年第12期2372-2376,共5页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立能有效分离18α-、18β-甘草酸的高效液相色谱方法并用于甘草酸二铵原料及制剂产品中异构体组成及相关杂质的控制。方法:采用C18色谱柱(4.6mm×250mm,5μm),以0.1mol·L-1高氯酸铵溶液(用氨水调节pH至8.0)-甲醇(48∶52)为流动相,检测波长250nm,柱温50℃,流速0.8mL·min-1。结果:18β-甘草酸与18α-甘草酸分离度不小于1.2,方法耐用性好;测得国内1个厂家3批甘草酸二铵原料及2个厂家各3批甘草酸二铵注射液中甘草酸二铵均为18α-、18β-2种构型的混合物,以18α-构型为主,18β-构型含量分别为27.4%～28.5%,27.2%～29.3%,20.4%～20.7%;样品中均存在EP5.0甘草酸单铵药品标准中收载的杂质A,发现杂质A也存在18α-、18β-2种构型,在上述色谱条件下得到有效分离。结论:甘草酸二铵产品为18α-、18β-两种构型的混合物,而国内现有药品标准方法不能分离18α-、18β-甘草酸,可采用本文中方法对其构型组成及相关杂质加以合理控制。Objective：To establish an HPLC method to segregate 18α-and 18β-glycyrrhizin to control isomer composition and related substance in diammonium glycyrrhizinate and its injection.Methods：The HPLC separation was carried out on a C18（4.6 mm×250 mm,5 μm）column at temperature of 50 ℃,using a UV detector setting detection wavelength 250 nm.Elution was carried out with 0.1 mol·L-1NH4ClO4（adjust to pH8.0 by ammonia solution）-methanol（48∶52）at a flow rate of 0.8 mL·min-1.Results：The resolution between 18α-and 18β-glycyrrhizin exceeded 1.2 and the method was robust.Determination of each three batches of diammonium glycyrrhizinate from one domestic pharmaceutical factory and each three batches of diammonium glycyrrhizinate injection from two domestic pharmaceutical factories showed that diammonium glycyrrhizinate were all comprised of 18α and 18β isomer,in which 18α was the main component and the content of 18β-isomer was 27.4%-28.5%,27.2%-29.3% and 20.4%-20.7% respectively.The impurity A which is controlled in the drug specification of ammonium glycyrrhizinate in EP 5.0 was also detected in all the samples and it was observed that the impurity A also had two isomers of 18α and 18β,which also could be separated efficiently using the above chromatogram method.Conclusion：Glycyrrhizin in its products always consists of 18α-and 18β-isomers but they can not be efficiently separated by available HPLC methods in domestic drug standards.The method above can be used for determination of 18α-and 18β-glycyrrhizin and related substances for glycyrrhizin products＇ quality control.

关 键 词：甘草酸二铵 18Α-甘草酸 18Β-甘草酸 甘草皂苷G2 异构体 高效液相色谱法 

分 类 号：R917[医药卫生—药物分析学]