HPLC测定罗布麻叶中总黄酮的含量

Determination of Flavonoids in the Leaves of Apocynum venetum by HPLC

摘要:

目的: 建立罗布麻叶中总黄酮的HPLC含量测定方法。 方法: 药材经提取水解后用 HPLC 通过测定其槲皮素和山奈酚的含量来确定总黄酮的含量,采用岛津 C₁₈色谱柱;甲醇-0.4％磷酸溶液(45 ∶40)为流动相;流速 1.0 mL ·min^-1;检测波长为 360 nm;柱温为 30 ℃; 结果: 槲皮素在 5.52~55.2 mg ·L^-1线性关系良好,r=0.999 9(n=6),平均回收率为 98.76%,RSD为 1.01%(n=6);山奈酚在 3.505~35.05 mg ·L^-1线性关系良好,r=0.999 9(n=6),平均回收率为 98.62%,RSD为 1.14%(n=6);3批样品中总黄酮含量为 17.08, 16.54,16.81 mg ·g^-1。 结论: 该方法准确可靠,重复性好,可以作为罗布麻叶总黄酮的含量测定方法。为完善罗布麻叶质量标准提供依据,同时为罗布麻叶的进一步研究奠定基础。

Abstract:

Objective: To establish a method for determination of total flavonoids in the leaves of Apocynum venetum by HPLC. Method: The total quercetrin and kaempferol content in the flavonoids was determined by HPLC after hydrolysis. The assay was performed on Shim-pack CLC-ODS column; methanol-0.4 % phosphoric acid (45 ∶40) was used as mobile phase; the flow rate was 1.0 mL ·min^-1, detection wavelength was at 360 nm, temperature of column was at 30 ℃. Result: Quercetrin was linear in the range of 5.52-55.2 mg ·L^-1 (r=0.999 9, n=6); the average recovery and relative standard deviation were 98.76% and 1.01% (n=6) respectively. Kaempferol was linear in the range of 3.505-35.05 g ·L^-1 (r=0.999 9,n=6); the average recovery and relative standard deviation were 98.62% and 1.14% (n=6) respectively. The content of total flavonoids in three samples was 17.08, 16.54 and 16.81 mg ·g^-1 respectively. Conclusion: This method was accurate and reliable. It can be used for the determination of the flavonoids in the leaves of A. venetum. It will provide the basis for the quality standards of Luobuma leaves and further study.