3种不同基原甘草中18α-甘草酸与18β-甘草酸的含量分析  

Simultaneous determination of 18 α-glycyrrhizic acid and 18 β-glycyrrhizic acid in three licorice samples from different origin by HPLC

作　　者：杨瑞[1] 李文东[2] 袁伯川 马永生[1] 周姗[1] 刘春生[1] 刘颖[1] 

机构地区：[1]北京中医药大学中药学院,北京100102 [2]北京市药品检验所,北京102206

出　　处：《药物分析杂志》2016年第6期1065-1071,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：北京市青年英才计划(YETP0819);国家自然科学基金(81503181)

摘　　要：目的:建立同时测定甘草中2种三萜类活性成分——18α-甘草酸与18β-甘草酸含量的HPLC分析方法,并对中国药典规定的3种不同基原甘草样品中18α-甘草酸与18β-甘草酸的含量进行分析比较。方法:以同一产地的3种不同基原2年生甘草作为实验样品;HPLC法对样品中的18α-甘草酸与18β-甘草酸进行测定。色谱柱:Agela Durashell C_(18)-AM(250 mm×4.6 mm,5μm);流速:0.8 mL·min^(-1);流动相:10 mmol·L^(-1)高氯酸铵水溶液(用氨水调节pH为8.2)-甲醇(48∶52);检测波长:250 nm;柱温:50℃。结果:在选择的色谱条件下,18α-甘草酸和18β-甘草酸达到了较好的分离;线性范围分别为0.010 70~0.214 0μg和0.216 4~4.328μg;检测限分别为3.210 ng和4.330 ng;定量限分别为11.26 ng和12.65 ng;平均回收率(n=3)分别为99.2%~100.1%(RSD≤0.93%)和99.8%~99.9%(RSD≤0.72%)。在3种基原的甘草样品中,光果甘草的18α-甘草酸与18β-甘草酸含量最高,分别为(2.650±0.064 21)mg·g^(-1)和(37.182±0.844 3)mg·g^(-1)(n=25);乌拉尔甘草的含量次之,分别为(1.975±0.057 12)mg·g^(-1)和(29.413±0.741 2)mg·g^(-1)(n=25);胀果甘草的含量最低,分别为(1.604±0.043 72)mg·g^(-1)和(17.810±0.502 1)mg·g^(-1)(n=25)。18α-甘草酸和18β-甘草酸的含量存在显著相关关系。结论:经方法学验证,本文方法可用于不同基原甘草中18α-甘草酸与18β-甘草酸的含量分析,并为以18α-甘草酸为目标的优质甘草筛选及定向育种奠定基础。Objective： To establish an HPLC method for simultaneous contents determination of 18 α- glycyrrhizic acid and 18 β -glycyrrhizic acid, which is a pair of epimer, and compare the different contents of these two triterpenoids in licorice stipulated in Chinese pharmacopoeia from three different origins. Methods： Three original plants of licorice which had been cultured for two years under the same condition were collected. The assay was performed through HPLC method. The contents were analyzed by using a chromatographic column Agela Durashell C18-AM （ 250 mm×4.6 mm,5μm ） at the column temperature 50 ℃. The mobile phase consisted of 10 mmol· L^-1 ammonium perchlorate water solution （ adjusted to pH 8.2 with ammonia water ） and methyl alcohol （ 48 ： 52 ） at the flow rate of 0.8 mL · min^-1, and the UV detection wavelength was set at 250 nm. Results：Under the selecglycyrrhizic acid could be well separated. The linearity ranges were 0.010 70-0.214 0 luted chromatographic conditions, 18 α-glycyrrhizic acid and 18β-g for 18 α -glycyrrhizic acid and 0.216 4-4.328 lug for 18β -glycyrrhizic acid. The LOD and LOQ for 18 α - glycyrrhizic acid were 3.210 ng and 11.26 ng, and the LOD and LOQ for 18β -glycyrrhizic acid were 4.330 ng and 12.65 ng. The average recoveries （ n=3 ） of 18 α-glycyrrhizic acid and 18 β-glycyrrhizic acid were 99.2%-100.1% （ RSD ≤ 0.93% ） and 99.8%-99.9% （ RSD ≤ 0.72% ）, respectively. It was demonstrated that the contents （ n=25 ） of 18 α -glycyrrhizic acid and 18 β -glycyrrhizic acid in Glycyrrhiza glabra L. were the highest, （ 2.650 ± 0.064 21 ） mg· g^-1 and （ 37.182 ± 0.844 3 ） mg·g^-1, respectively; the contents （ n=25 ） of 18 a-glycyrrhizic acid and 18 13-glycyrrhizic acid in Glycyrrhiza uralensis Fisch. were the second, （ 1.975 ± 0.057 12 ） mg· g^-1 and （ 29.413 ± 0.741 2 ） mg· g^-1 respectively; and the contents （ n=25 ） of 18 α - glycyrrhizic acid and 18 β -glycyrrhizic acid in Glycyrrhiza inflata Bat. were

关 键 词：不同基原比较 甘草 乌拉尔甘草 光果甘草 胀果甘草 甘草酸 三萜类活性成分 差向异构体 HPLC 

分 类 号：R917[医药卫生—药物分析学]