高效液相色谱法测定四乙酰葛根素的含量及有关物质

Determination of Content and Related Substances of Tetra-Acetylated Puerarin by HPLC

目的建立测定四乙酰葛根素的含量及有关物质的高效液相色谱法。方法采用Agilent XDB-C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-水(45∶55),流速为1.2 mL/min,检测波长为250 nm,柱温为30℃。结果四乙酰葛根素的质量浓度在0.103 8～0.934 2μg范围内与峰面积线性关系良好,Y=12 871 X-24.554,r=0.999 9(n=7),检测限为6.65 ng/mL(S/N=3);平均加样回收率为99.94%,RSD=1.32%(n=9),有关物质含量为1.02%～1.28%。结论该方法简便、准确,可用于四乙酰葛根素的含量测定。

Objective To establish an HPLC method for determination of the content and related substances of tetra- acetylated puerarin （TAP）. Methods Agilent XDB- C18 chromatographic column（250 mm ×4. 6 mm,5 μm）was used and the mobile phase consisted of acetonitrile-water（45：55） with a flow rate of 1.2 mL/min. The UV detection wavelength was set at 250 nm. The column temperature was controlled at 30℃. Results The calibration curve was linear within the range of 0. 103 8- 0. 934 2μg, Y= 12 871X-24. 554, r = 0. 999 9 （ n = 7）. The detective limit was 6.65 ng/mL（ S/N = 3）. The mean recoveries was 99.94% （ RSD = 1.32% , n = 9）. The content of related substances of TAP was in the range of 1.02% -1.28%. Conclusion The method is simple,accurate and is applicable for the quality control of TAP.