针对中国国家药品标准(WS1-XG-2002)中,甘草酸单铵盐含量测定方法分析时间长、效率低、不能实现对甘草酸主成分异构体18α-甘草酸(18α-Gly)和18β-甘草酸(18β-Gly)的有效分离,以及未对有关物质进行准确定量分析和质量控制等问题,改进并建立甘草酸单铵盐原料药及不同剂型制剂中有关物质及含量的检测方法.对色谱条件进行优化,采用Durashell-C18柱(4.6 mm×250 mm,5μm),以0.01 mol/L高氯酸铵(氨水调节pH至8.20)-甲醇(48∶52,V/V)为流动相,流速0.80 mL/min,检测波长254 nm,柱温50℃,进样量10 μL.18α-Gly,18β-Gly,甘草酸单铵盐杂质A和杂质B在0.5～100.0 μg/mL内线性关系良好(r=0.999 9),检出限分别为0.10,0.10,0.15,0.15μg/mL,平均回收率(n=3)为98.06％～101.61％.与中国国家药品标准(WS1-XG-2002)收载的甘草酸类制剂质量标准比较,本实验建立的方法能够实现对主成分异构体的有效分离,并能真实、准确地检测各有关物质的含量,方法精密度,重现性良好,灵敏度高,结果准确可靠,可用于甘草酸单铵盐原料药及其不同剂型制剂的检测分析及质量控制.
[1] 梁玉玲, 管延英, 阳丽, 王淑敏, LIANG Yu-ling, GUAN Yan-ying, YANG Li, WANG Shu-min. 胀果甘草愈伤组织培养及甘草酸含量分析 [J]. 河北大学学报（自然科学版） 2000.doi:10.3969/j.issn.1000-1565.2000.04.016
[2] 阎正, 肖进进, 崔建波, 张健申, YAN Zheng, XIAO Jin-jin, CUI Jian-bo, ZHANG Jian-shen. 顶空毛细管气相色谱法测定甘草酸二铵中乙醇残留量 [J]. 河北大学学报（自然科学版） 2005.doi:10.3969/j.issn.1000-1565.2005.06.013
[3] 曹站霞, 王纪力, 唐进法. HPLC测定复方甘草酸苷氯化钠注射液中甘草酸胺含量 [J]. 中成药 2009.
[4] CLASSEN-HOUBEN D, SCHUSTER D, DA CUNHA T. Selective inhibition of 11Beta-hydroxysteroid dehydrogenase 1 by 18alpha-glycyrrhetinic acid but not 18beta-glycyrrhetinic acid [J]. J Steroid Biochem Mol Biol 2009, 113(3-5).
[5] Lin JC, Cherng JM, Hung MS, Baltina LA, Baltina L, Kondratenko R. Inhibitory effects of some derivatives of glycyrrhizic acid against Epstein-Barr virus infection: Structure-activity relationships. [J]. Antiviral Research 2008, 1(1).
[6] Ren, W., Lu, Y., Jing, J., Zhu, J., Wan, D., Zhao, D., Zheng, J., Fang, F., Hea, Y., Chen, X.. Determination of glycyrrhizin in dog plasma by liquid chromatography-mass spectrometry and its application in pharmacokinetic studies [J]. European journal of mass spectrometry 2010, 6(6).
[7] 王宇光, 杨明会, 马增春, 梁乾德, 谭洪玲, 肖成荣, 高月, WANG Yuguang, YANG Minghui, MA Zengchun, LIANG Qiande, TAN Hongling, XIAO Chengrong, GAO Yue. 18β-甘草酸和18α-甘草酸对大鼠原代肝细胞CYP3A酶表达的影响 [J]. 中国中药杂志 2009.
[8] ZOU Qiaogen, WEI Ping, LI Jing. Simultaneous determination of 18α-and 18β-glycyrrhetic acid in human plasma by LC-ESI-MS and its application to pharmacokinetics [J]. Biomedical Chromatography 2009, 23(01).
[9] 赵鸿, 鹿程华, 玄春香. 紫外分光光度法测定甘草酸单铵含量 [J]. 中国医院药学杂志 2006.doi:10.3321/j.issn:1001-5213.2006.12.065
[10] 沈阳, 沈金灿, 庄峙厦, 王小如. HPCE法与HPLC法用于甘草中主要活性成分甘草酸及其降解产物甘草次酸分析的比较 [J]. 2006.
[11] Sabbioni C, Mandrioli R, Ferranti A, Bugamelli F, Saracino MA, Forti GC, Fanali S, Raggi MA. Separation and analysis of glycyrrhizin, 18 beta-glycyrrhetic acid and 18 alpha-glycyrrhetic acid in liquorice roots by means of capillary zone electrophoresis [J]. Journal of chromatography, A: Including electrophoresis and other separation methods 2005, 1(1).
[12] SAKAE AMAGAYA, ETSUKO SUGISHITA, YUKIO OGIHARA. Separation and quantitative analysis of 18α-glycyrrhetinic and 18β-glycyrrhetinic acid in glycyrrhizae Radix by gas-liquid chromatography [J]. Journal of Chromatography 1985, 320.
[13] 吕坚, 吴锡铭, 周晴. 气相色谱法测定大鼠血浆中18-H甘草酸差向异构体含量 [J]. 中国药学杂志 1993, 28(09).
[14] 宋红波, 刘茜, 赵辉, 张丽英, 多凯, 赵英明, 杨毅文. 甘草酸铵类制剂中甘草酸差向异构体的分析 [J]. 2012.
[15] Qiaogen Zou, Ying Zhan, Ping Wei. Isolation and Characterization of Related Impurities of Monoammonium Glycyrrhizinate and Development of RP-HPLC Method for Quality Control Study [J]. Journal of liquid chromatography and related technologies 2009, 13/16(13/16).
[16] Shen SF, Chang ZD, Liu J, Sun XH, Hu X, Liu HZ. Simultaneous determination of glycyrrhizic acid and liquiritin in Glycyrrhiza uralensis extract by HPLC with ELSD detection [J]. Journal of liquid chromatography and related technologies 2006, 13/16(13/16).
[17] 颜苗, 张金娇, 李焕德, 朱荣华, 李兰芳. RP-HPLC法同时测定甘草酸制剂中18α -、18β-甘草酸的含量 [J]. 2012.
