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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 determination of trace elements in monoammonium glycyrrhizinate by high pressure solid phase extraction coupled with high performance liquid chromatography-mass spectrometry. Methods: High performance liquid chromatography (HPLC) was used as an auxiliary method to determine the best method of high pressure solid phase extraction by optimizing the conditions. Using a 5C18-MS-Ⅱ column (20.0 mm × 20.0 mm) as the extraction column, 0.5 mL of the sample was injected into the column and eluted at 16 mL · min-1 with 35% acetonitrile-acetic acid solution (pH 2.2) The elution was carried out at a flow rate of 4.5 min. The resulting extract was subjected to liquid chromatography-mass spectrometry (LC-MS) analysis 100-fold concentration. Results: Ten chemical constituents of monoammonium glycyrrhizinate were rapidly identified under the optimized conditions of high pressure solid-phase extraction coupled with liquid chromatography-mass spectrometry (MS-MS). Among them, six kinds of trace components 3-O- [β-D-glucuronic acid-β-D-glucuronic acid] -30-O-β-D-celery glycyrrhetinic acid / 3-O- [β -D-glucuronic acid-β-D-glucuronic acid] -30-O-β-D-arabinose glycyrrhetinic acid, glycyrrhizin F3,18α-glycyrrhizin G2,3-O- [β-D - rhamnose] -24-hydroxyglycyrrhizin, glycyrrhizin J2, glycyrrhizin B2; by liquid chromatography, multi-stage mass spectrometry and UV spectral information, compared with the control, confirming the four main chemical raw materials glycyrrhizic acid Ingredients were glycyrrhizin G2,18β-glycyrrhizic acid, ural glycyrrhizin B, 18α-glycyrrhizinic acid. Conclusion: This method can quickly and effectively identify the chemical components in the raw materials of mono glycyrrhizic acid without reference substance, and provide a basis for the quality control and safe application of monoammonium glycyrrhizinate-related products.