杨学东;唐绪岩;桑琳;
YANG Xue-dong,TANG Xu-yan,SANG Lin(School of Pharmaceutical Science and Technology,Tianjin University,Tianjin 300072,China)

• 1:天津大学药物科学与技术学院

摘要(Abstract)：

目的:建立高压固相萃取-高效液相色谱-质谱法快速鉴定甘草酸单铵原料中微量成分的方法。方法:以高效液相制备色谱仪为辅助手段,通过条件优化,确定高压固相萃取的最佳方法。以5C18-MS-Ⅱ色谱柱(20.0 mm×20.0 mm)为萃取柱,进样0.5 mL,以35%乙腈-醋酸溶液(pH 2.2)为洗脱剂,在16 mL·min-1的洗脱流速下进行洗脱,萃取周期为4.5 min。所得萃取液在浓缩100倍后进行液相色谱-质谱(LC-MS)分析。结果:在最优化的高压固相萃取-液相色谱-质谱条件下,快速鉴定出甘草酸单铵原料中的10种化学成分,其中采用多级质谱信息分析和文献对照推测出6种微量成分的化学结构,分别为3-O-[β-D-葡糖醛酸-β-D-葡糖醛酸]-30-O-β-D-芹菜糖甘草次酸/3-O-[β-D-葡糖醛酸-β-D-葡糖醛酸]-30-O-β-D-阿拉伯糖甘草次酸,甘草皂苷F3,18α-甘草皂苷G2,3-O-[β-D-鼠李糖]-24-羟基甘草酸,甘草皂苷J2,甘草皂苷B2;通过液相色谱、多级质谱和紫外光谱信息,与对照品对照,确证了甘草酸单铵原料中4种主要化学成分,分别为甘草皂苷G2,18β-甘草酸,乌拉尔甘草皂苷乙,18α-甘草酸。结论:该方法无需对照品即可快速有效地鉴定出甘草酸单铵原料中的化学成分,为甘草酸单铵相关产品的质量控制和安全应用提供了依据。
Objective: To establish a method for rapid identification of micro-constituents in monoammonium glycyrrhizinate by high-pressure solid phase extraction-high performance liquid chromatography-mass spectrometry.Method: HPLC preparative chromatograph was adopted for determining the optimal method for high-pressure solid phase extraction under optimal conditions.5C18-MS-Ⅱ column(20.0 mm×20.0 mm) was used as the extraction column,with 35% acetonitrile-acetic acid solution(pH 2.20) as eluent at the speed of 16 mL·min-1.The sample size was 0.5 mL,and the extraction cycle was 4.5 min.Then,extract liquid was analyzed by high performance liquid chromatography-mass spectrometry(HPLC-MS) after being concentrated by 100 times.Result: Under the optimal condition of high-pressure solid phase extraction-high performance liquid chromatography-mass spectrometry,10 components were rapidly identified from monoammonium glycyrrhizinate raw materials.Among them,the chemical structures of six micro-constituents were identified as 3-O-[β-D-glucuronopyranosyl-β-D-glucuronopyranosyl]-30-O-β-D-apiopyranosylglycyrrhetic/3-O-[β-D-glucuronopyranosyl-β-D-glucuronopyranosyl]-30-O-β-D-arabinopyranosylglycyrrhetic,glycyrrhizic saponin F3,22-hydroxyglycyrrhizin/18α-glycyrrhizic saponin G2,3-O-[β-D-rhamnopyranosyl]-24-hydroxyglycyrrhizin,glycyrrhizic saponin J2,and glycyrrhizic saponin B2 by MSn spectra analysis and reference to literatures.Four main chemical components were identified as glycyrrhizic saponin G2,18β-glycyrrhizic acid,uralglycyrrhizic saponin B and 18α-glycyrrhizic acid by liquid chromatography,MSn and ultraviolet spectra information and comparison with reference substances.Conclusion: The method can be used to identify chemical constituents in monoammonium glycyrrhizinate quickly and effectively,without any reference substance,which provides basis for quality control and safe application of monoammonium glycyrrhizinate-related products.

关键词(KeyWords)： 甘草酸单铵;微量成分;快速鉴定;高压固相萃取;LC-MS
monoammonium glycyrrhizinate;micro-constituent;rapid identification;high-pressure solid phase extraction;liquid chromatography-mass spectrometry

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 杨学东;唐绪岩;桑琳;
YANG Xue-dong,TANG Xu-yan,SANG Lin(School of Pharmaceutical Science and Technology,Tianjin University,Tianjin 300072,China)

Email:

DOI:

参考文献(References)：

• [1]Zapesochnaya G G,Zvonkova E N,Kurkin V A,et al.Someproperties of glycyrrhizic acid[J].Chem Nat Compd,1994,30(6):720.
• [2]沈阳,沈金灿,庄峙厦,等.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].药物分析杂志,2006,26(4):421.
• [3]佟连生,田云.从甘草酸粗品制取甘草酸单铵盐[J].黑龙江医药科学,2004,27(4):51.
• [4]梁月琴,刘琴棣,吴德政.高效液相色谱法测定甘草及其制剂中甘草酸含量[J].医药研究通讯,1997,26(8):33.
• [5]木其耳,刘有平,李萌.甘草酸单铵盐中有关物质的LC-MSn研究[J].中国医药工业杂志,2009,40(11):844.
• [6]Zou Q G,Zhang Y,Wei P,et al.Isolation and characterizationof related impurities of monoammonium glycyrrhizinate and devel-oped of RP-HPLC method for quality control study[J].J LiqChromatogr Related Technol,2009,32(13):1953.
• [7]李文,沙沂,陈丽霞.甘草酸二铵18位差向异构体的核磁共振研究[J].沈阳药科大学学报,2005,22(4):273.
• [8]吴锡铭,吕坚,茹仁萍.甘草酸18H差向异构体的比较研究[J].中国药学杂志,1993,28(4):215.
• [9]Sabbioni C,Mandrioli R,Ferranti A,et al.Seperation and anal-ysis of glycyrrhizin,18β-glycyrrhetic acid and 18α-glycyrrheticacid in liquorice roots by means of capillary zone electrophoresis[J].J Chromatogr A,2005,1081(1):65.
• [10]屈景年,罗炳初,匡云飞,等.甘草酸氨基酸复盐的制备及性质[J].化学世界,2007(9):525.
• [11]Kitagawa I,Hori K,Taniyama T,et al.Licorice root.A naturalsweetener and an important ingredient in Chinese medicine[J].Chem Pharm Bull,1993,41(8):1337.
• [12]周燕,王明奎,廖循,等.甘草化学成分的高效液相色谱-串联质谱分析[J].分析化学,2004,32(2):174.