RP-HPLC法同时测定不同产地甘草中9个主要成分的含量

Simultaneous determination of nine constituents in the roots and rhizomes of Glycyrrhiza uralensis from different producing areas by RP-HPLC

目的:建立同时测定甘草药材中芹糖基甘草苷、甘草苷、芹糖基异甘草苷、异甘草苷、甘草查尔酮B、甘草素、刺甘草查尔酮、异甘草素和甘草酸含量的高效液相色谱方法。方法:采用DiamonsilTMC18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.05%磷酸水溶液为流动相,梯度洗脱,流速为0.8 mL.min-1,检测波长为276 nm(0～20 min)、360 nm(20～30 min)、276 nm(30～35 min)、370 nm(35～48 min)、248 nm(48～55 min),柱温为40℃。结果:芹糖基甘草苷、甘草苷、芹糖基异甘草苷、异甘草苷、甘草查尔酮B、甘草素、刺甘草查尔酮、异甘草素和甘草酸的浓度分别在13.1-262μg.mL-1(r=0.9994)、8.3-100μg.mL-1(r=0.9997)、5.2～104μg.mL-1(r=0.9997)、0.96～11.5μg.mL-1(r=0.9995)、0.31～10.00μg.mL-1(r=0.9999)、1.11～22.2μg.mL-1(r=0.9991)、0.23～7.25μg.mL-1(r=0.9998)、0.52～10.40μg.mL-1(r=0.9996)、25.25～808.0μg.mL-1(r=0.9995)与各自峰面积值呈良好的线性关系;平均回收率(n=6)分别为103.6%、104.5%、104.9%、100.4%、102.2%、101.5%、100.4%、98.5%、101.7%。结论:本方法简便,重复性好,可以更好地用于甘草药材的质量控制。

Objective：To develop an HPLC method for simultaneous determination of 9 main constituents （liquiritin apioside, liquiritin, isoliquiritin apioside, isoliquiritin, licochalcone B, liquiritigenin, echinatin, isoliquiritigenin, glycyr- rhizic acid）in the roots and rhizomes of Glycyrrhiza uralensis Fisch. Methods ：The analyses were performed on a Dia- monsil^TM ODS C18 column （250 mm× 4. 6 mm,5 p.m）with the gradient elution of aeetonitrile- 0. 05% phosphoric acid aqueous at a flow rate of 0. 8 mL · rain^-1. The detection wavelength was set at 276 nm（0 -20 min） ,360 nm（20 - 30 min）,276 nm （30 - 35 rain）, 370 nm （35 - 48 rain）, 248 nm （48 - 55 min） and the column temperature was at 40 ℃. Results： Good linearity was obtained for liquiritin apioside,liquiritin,isoliquiritin apioside,isoliquiritin,licochal- cone B, liquiritigenin, echinatin, isoliquiritigenin, and glycyrrhizic acid in ranges of 13.1 - 262 μg · mL^-1 （ r = 0.9994） ,8.3 - 100 μg · mL^-1 （r =0.9997）,5.2 - 104 μg · mL^-1 （r =0.9997）,0.96 - 11.5μg ·mL^-1 （r = 0. 9995 ）,0. 31 - 10. 00 μg · mL^- 1 （ r = 0. 9999）, 1.11 - 22. 2 μg · mL^- 1 （ r = 0. 9991 ）, O. 23 - 7. 25 μg · mL ^- 1 （ r = 0. 9998 ） ,0. 52 - 10. 40 μg· mL^- 1 （r = 0. 9996）and 25.25 -808.0μg · mL^- 1 （r = 0. 9995 ）, respectively. The average recovery （ n = 6） of the nine constitutions was 103.6%, 104. 5%, 104. 9%, 100. 4%, 102. 2%, 101.5%, 100. 4%, 98.5% ,101.7% ,respectively. Conclusion：The developed HPLC method is simple and repeatable, which is helpful for the quality control of the roots and rhizomes of G. uralensis.