黄芩苷铝HPLC检测方法的建立

Establishment of HPLC determination method for baicalin aluminum

为了探究测定黄芩苷铝含量的有效方法,试验先摸索了供试品前处理方法,将黄芩苷铝完全转化成黄芩苷后以黄芩苷为对照品,建立测定黄芩苷含量的高效液相色谱(HPLC)检测方法,绘制标准曲线,并考察该方法的系统适用性、准确度、精密度、重复性、耐用性和稳定性,最后利用该方法对样品含量进行检测。结果表明:供试品前处理条件为向黄芩苷铝中加入乙腈-0.05%磷酸溶液(40∶60)并超声30 min;液相色谱条件,色谱柱为Hypersil ODS色谱柱(150 mm×4.6 mm, 5μm),流动相为乙腈-0.05%磷酸水溶液(30∶70),检测波长为278 nm,流速为1.00 mL/min,柱温为30℃,进样量为10μL。在该条件下,黄芩苷在20~320μg/mL内线性关系良好(R^(2)=0.999 9),精密度高、重复性好,平均回收率为101.24%,供试品溶液在36 h内稳定。在测试范围内,柱温、流速和色谱柱对样品中黄芩苷的含量测定无影响。说明建立的黄芩苷铝HPLC检测方法操作简单,可靠性高,可以用作黄芩苷铝含量的测定。

In order to explore the effective method for the determination of baicalin aluminum content, in the experiment, firstly, the pretreatment method of the test sample was explored;the high-performance liquid chromatography(HPLC) detection method for the determination of baicalin content was established after the aluminum baicalin was completely converted into baicalin and the baicalin was used as the reference substance;the standard curve was drawn, and the system applicability, accuracy, precision, repeatability, durability and stability of the method were investigated, and finally the sample content was detected by the method. The results showed that the pretreatment conditions of the test samples were acetonitrile-0.05% phosphoric acid solution(40∶60) to baicalin aluminum and sonication for 30 min;liquid chromatography conditions were Hypersil ODS column(150 mm × 4.6 mm, 5 μm);mobile phase was acetonitrile-0.05% phosphoric acid aqueous solution(30∶70);the detection wavelength was 278 nm;the flow rate was 1.00 mL/min;the column temperature was 30 ℃;and the injection volume was 10 μL. Under these conditions, baicalin had a good linear relationship(R^(2)=0.999 9) in 20-320 μg/mL, high precision and good repeatability;the average recovery rate was 101.24%, and the test solution was stable within 36 h. Within the test range, column temperature, flow rate, and column had no effect on the determination of baicalin content in the sample. The results indicated that the established HPLC method for the detection of baicalin aluminum was simple to operate and had high reliability, and could be used as the determination of the content of baicalin aluminum.