微乳液相色谱法同时测定大黄中5种蒽醌类衍生物的含量

Content Determination of 5 Anthraquinone Derivatives in Rhei Radix Et Rhizoma by Microemulsion Liquid Chromatography

目的:建立同时测定大黄中芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚含量的方法。方法:采用微乳液相色谱法。以微乳为流动相,通过对影响分离度和保留时间的因素进行考察,优化最佳微乳体系组成为2.5%(W/V)十二烷基硫酸钠-0.1%(V/V)正辛烷-8.0%(V/V)正丁醇-0.5%(V/V)三乙胺(磷酸调pH值至3.0);色谱柱为HypersilODS2(250mm×4.6mm,5μm),柱温为28℃,流速为1mL·min-1,检测波长为254nm。结果:芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚的进样浓度线性范围分别为4.7～47.0μg·mL-1(r=0.9999)、5.1～51.0μg·mL-1(r=0.9996)、5.5～55.0μg·mL-1(r=0.9996)、6.3～63.0μg·mL-1(r=0.9999)和0.4～40.0μg·mL-1(r=0.9987);五者平均加样回收率分别为98.67%、97.53%、101.37%、99.34%和99.52%,RSD(n=6)分别为0.31%、0.38%、0.49%、0.53%和0.87%。结论:本方法简便、准确,可用于大黄中5种蒽醌类衍生物的含量测定。

OBJECTIVE： To develop the method for the content determination of aloe-emodim, rhein, emodin, chrysophanol and physcione. METHODS： Microemulsion liquid chromatography （MELC） was adopted, and mobile phase was microemulsion that was optimized by inspecting the influence factors of separation and retention time. The optimized microemulsion was 2.5 % （W/ V） sodium dodecyl sulfate-0.1% （V/V） n-octane-8.0 % （V/V） n-butanol-0.5 % （V/V） triethylamine （pH adjusted to 3.0 by phosphoric acid）. Hypersil ODS2 （250 mmx4.6 mm, 5 ktm） column was selected with column temperature of 28 ℃ at a flow rate of 1 mL· min^-1, and the detection wavelength was set at 254 nm.RESULTS：The linear ranges were 4.7~47.0 μg·mL^-1 for aloe-emodim （r= 0.999 9）, 5.1-51.0μg·mL^-1 for rhein（r=0.999 6）, 5.5-55.0μg·mL^-1 for emodin （r=0.999 6）, 6.3-63.0 μg·mL^-1 for chrysophanol （r=0.999 9）, 0.4-40.0μg·mL^-1 for physcione （r=0.998 7）. The average recoveries were 98.67% , 97.53% , 101.37% , 99.34% and 99.52%, respectively. RSDs were 0.31%, 0.38%, 0.49%, 0.53% and 0.87% （n=6）. CONCLUSION： The method is simple and accurate, and can be used for the content determination of 5 anthraquinone derivatives in Rhei Radix Et Rhizoma.