青叶胆高效液相色谱特征图谱研究及指标性成分的含量测定

Study on the specific chrometogram of Swertia mileensis and determination of target components by HPLC

目的:建立青叶胆高效液相特征图谱的测定方法,并对其中獐牙菜苦苷、龙胆苦苷、獐牙菜苷3种指标性成分进行含量测定。方法:采用Phenomsil C18色谱柱(250mm×4.6mm,5μm),流动相为甲醇-乙腈-0.02%磷酸,梯度洗脱,流速1.0mL.min-1,柱温35℃,检测波长254nm,记录时间70min,对10批青叶胆药材的特征图谱进行研究及含量测定。结果:初步建立了青叶胆药材的高效液相特征图谱,标定了14个共有特征峰。实验中各共有峰相对保留时间的RSD小于5%,相对峰面积的相似度在80%左右。待测的3种成分均达到基线分离,獐牙菜苦苷、龙胆苦苷、獐牙菜苷的线性范围分别为0.33～1.98μg(r=0.9997)、0.16～0.96μg(r=0.9997)、0.20～1.20μg(r=0.9999),回收率分别为99.9%,98.2%,100.7%。结论:此方法稳定、准确、有效,可以全面的反映青叶胆药材的质量,可作为控制青叶胆药材质量的依据。

OBJECTIVE To establish specific chromatogram of swertia mileensis by using HPLC for quality evalution and to determinate the contents of therr iridoids, swertiamarin , gentiopicroside, and sweroside in swertia mileensis. METHODS The analysis was performed on a Phenomsil Ca8 column （4. 6 mm × 250 mm, 5μm） with methanol- acetonitrile- water （0. 02% phosphoric acid） as mobile phase in a gradient mode at a flow rate of 1.0 mL·min^-1 ,and at a column temperature of 35 ℃. The detection wavelength was at 254 nm. RESULTS The specific chromatogram of Sweitia mileensis was established and 14 common peaks were indicated. Through reference substances,3 common peak were identified. The relative standard deviations for relative retention values and relative peak areas were less than 5% in the precision and repeated test. Base line resolution was achieved. The linear ranges of swertiamarin ,gentiopicroside and sweroside were 0. 33-1.98 μg（r= 0. 999 7）,0. 16-0. 96μg（r= 0. 999 7）, 0. 20-1.20 μg（r=0. 999 9）, respectively. The average recoveries were 99. 9% , 98. 2%, 100. 7%, respectively CONCLUSION The method is accurate.feasible and reliable, and can be used for the quality of Swertia mileensis.