替代对照品法测定甘草流浸膏中甘草苷和甘草酸含量  

Content determination of liquiritin and glycyrrhizic acid in Licorice Liquid Extract by substitute reference substance method

作　　者：刘航 魏长勇 余敏灵 LIU Hang;WEI ChangYong;YU Minling(Leshan Municipal Food and Drug Inspection and Testing Center,Leshan,Sichuan 614000,China)

机构地区：[1]乐山市食品药品检验检测中心,四川乐山614000

出　　处：《现代医药卫生》2021年第1期47-51,共5页Journal of Modern Medicine & Health

摘　　要：目的建立以橙皮苷为替代对照品测定甘草流浸膏中甘草苷和甘草酸含量的高效液相色谱法。方法采用替代对照品内加入法,以橙皮苷为替代对照品,通过测定甘草酸铵与橙皮苷、甘草苷与橙皮苷的相对校正因子(f值),利用f值计算甘草流浸膏中甘草苷和甘草酸含量。色谱条件:Welch Ultimate C 18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.1%三乙胺溶液(用磷酸调pH值为2.5)为流动相,梯度洗脱,检测波长为284 nm(橙皮苷)、276 nm(甘草苷)、252 nm(甘草酸铵),柱温为30℃,流速为1.0 mL/min。结果橙皮苷、甘草苷、甘草酸铵进样量分别为0.0332~0.4983、0.0493~0.7402、0.4330~6.4951μg,与其峰面积具有良好的线性关系(r=0.9995、0.9992、0.9993);甘草苷和甘草酸铵与橙皮苷的f值分别为1.0814、0.4492;甘草苷和甘草酸的平均回收率分别为98.96%、97.67%。替代对照品法样品含量测定数据与外标法比较,差异无统计学意义(P>0.05)。结论以橙皮苷为对照品,测定甘草流浸膏中甘草苷和甘草酸含量的高效液相色谱法具有成本低、实验时间短、操作过程和仪器噪音影响小等特点。Objective To establish a HPLC method for the determination of liquiritin and glycyrrhizic acid contents in Licorice Liquid Extract by using hesperidin as substitute reference substance.Methods The substitute reference substance internal addition method was used and hesperidin served as the substitute reference substance.The relative correction factors(f value)of ammonium glycyrrhizinate to hesperidin and liquiritin to hesperidin were measured,and the f values were used to calculate the content of glycyrrhizic acid and liquiritin in Licorice Liquid Extract.The chromatographic conditions:the Welch Ultimate C 18 chromatographic column(250 mm×4.6 mm,5μm)was used,acetonitrile-0.1%triethylamine solution(adjusting the pH value to 2.5 with phosphoric acid)as the mobile phase,gradient elution,the detection wavelength:284 nm(hesperidin),276 nm(liquiritin)and 252 nm(ammonium glycyrrhizinate),the column temperature 30℃,and the flow rate 1.0 mL/min.Results The sample amounts of liquirtin,hesperidin and ammonium glycyrrhizinate were in the range of 0.0332-0.4983,0.0493-0.7402,0.4330-6.4951μg,which showed the good linear relationship with the peak area(r=0.9995,0.9992,0.9993).The f values of liquiritin and ammonium glycyrrhizinate to hesperidin were 1.0814 and 0.4492,the average recovery rates of liquiritin and glycyrrhizic acid were 98.96%and 97.67%,respectively.Comparing the contents measurement data by the substitute reference substance method with the external standard method,the difference was not statistically significant(P>0.05).Conclusion With hesperidin as the reference substance,the HPLC method for the determination of liquiritin and ammonium glycyrrhizinate contents in Licorice Liquid Extract has the characteristics of low cost,short experimental time and little influence of operation process and instrument noise.

关 键 词：高效液相色谱法 甘草酸 替代对照品 内加入法 甘草流浸膏 

分 类 号：R917[医药卫生—药物分析学] R446[医药卫生—药学]