HPLC法同时测定龙胆泻肝丸中8个活性成分的含量

Simultaneous determination of eight active components in Longdan Xiegan pills by HPLC

目的:建立同时测定龙胆泻肝丸中龙胆苦苷、獐牙菜苦苷、京尼平苷、京尼平龙胆双糖苷、黄芩苷、汉黄芩苷、黄芩素和汉黄芩素含量的高效液相色谱法。方法:采用Sun FireTM C18色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈(A)-0.1%磷酸水(B),梯度洗脱(0~6 min,10%A→15%A;6~7 min,15%A;7~10 min,15%A→16%A;10~15 min,16%A;15~20 min,16%A→30%A;20~35 min,30%A→33%A;35~38 min,33%A→38%A;38~60 min,38%A→48%A),流速为1.0 m L·min^(-1),检测波长为254 nm,柱温为30℃。结果:龙胆苦苷、獐牙菜苦苷、京尼平苷、京尼平龙胆双糖苷、黄芩苷、汉黄芩苷、黄芩素、汉黄芩素质量浓度分别在2.5~100μg·m L^(-1)(r=0.999 6)、2.5~100μg·m L^(-1)(r=0.999 6)、5~200μg·m L^(-1)(r=0.999 6)、2.5~100μg·m L^(-1)(r=0.999 7)、5~200μg·m L^(-1)(r=0.999 5)、2.5~100μg·m L^(-1)(r=0.999 6)、1.25~50μg·m L^(-1)(r=0.999 8)和1.25~50μg·m L^(-1)(r=0.999 8)范围内与峰面积呈良好的线性关系;平均回收率(n=9)分别为99.4%、99.2%、99.3%、98.7%、99.6%、98.7%、98.2%和98.0%。6批样品中上述8个成分的含量范围分别为龙胆苦苷1.700~1.717 mg·g^(-1),獐牙菜苦苷0.524~0.544 mg·g^(-1),京尼平苷3.674~3.883 mg·g^(-1),京尼平龙胆双糖苷1.695~1.701 mg·g^(-1),黄芩苷7.235~7.440 mg·g^(-1),汉黄芩苷1.233~1.275 mg·g^(-1),黄芩素0.514~0.523 mg·g^(-1),汉黄芩素0.294~0.298 mg·g^(-1)。结论:该方法可用于龙胆泻肝丸的质量控制。

Objective： To develop an HPLC method for simultaneous determination of gentiopicroside, swertiamarin, geniposide, genipingentiobioside, baicalin, wogonoside, haicalein and wogonin in Longdan Xiegan pills. Methods： The chromatographic separation was performed on a SunFireTM C18 column （ 4.6 mm ×250 mm, 5 μm ）with gradient elution （ 0-6 min, 10%A → 15%A; 6-7 min, 15%A; 7-10 min, 15%A → 16%A; 10- 15 min, 16%A; 15-20 min, 16%A → 30%A; 20-35 min, 30%A → 33%A; 35-38 min, 33%A → 38%A; 38- 60 min, 38%A → 48%A ）of acetonitrile （ A ）and 0.1% phosphoric acid （ B ）ataflow rate of 1.0 mL·min-1. The detecting wavelength was set at 254 nm and the column temperature was maintained at 30℃. Results： The calibration curves were linear within the range of 2.5-100μg·mL-1 （ r=0.999 6 ） for gentiopicroside, 2.5-100 μg·mL-1 （ r=0.999 6 ）for swertiamarin, 5-200 μg·mL-1 （ r=0.999 6 ）for geniposide, 2.5-100 μg·mL-1（ r=0.999 7 ）for genipingentiobioside, 5-200μg·mL-1（ r=0.999 5 ） for baicalin, 2.5-100 μg·mL-1 （ r=0.999 6 ） for wogonoside, 1.25-50μg·mL-1 （ r=0.999 8 ） for baicalein and 1.25-50 μg·mL-1 （ r=0.999 8 ） for wogonin. The recoveries （ n=9 ） of gentiopicroside, swertiamarin, geniposide, genipingentiobioside, baicalin, wogonoside, baicalein and wogonin were 99.4%, 99.2%, 99,3%, 98.7%, 99.6%, 98.7%, 98.2%and 98.0%, respectively. The content ranges of eight components in six batches were 1.700-1.717mg·g-1 for gentiopicroside, 0.524-0.544mg·g-1 for swertiamarin, 3.674-3.883 mg·g-1 for geniposide, 1.695-1.701mg·g-1 for genipingentiobioside, 7.235-7.440mg·g-1 for baicalin, 1.233-1.275mg·g-1 for wogonoside, 0.514-0.523mg·g-1 for baicalein and 0.294-0.298mg·g-1 for wogonin. Conclusion： The method can be used for the quality control of Longdan Xiegan pills.