HPLC法测定鞣酸蛋白原料及制剂中鞣酸的含量

Determination of Tannic Acid in Albumin Tannate and Its Preparations by HPLC

目的：建立HPLC法测定鞣酸蛋白原料及制剂中鞣酸的含量。方法：使用Techmate C18-ST（4.6 mm ×250 mm，5μm）色谱柱；流动相A为0.5％冰醋酸溶液，流动相B为乙腈，流量0.8 mL· min^-1，梯度洗脱；柱温30.0℃；检测波长272 nm；进样量10μL；使用SPSS19.0软件设计正交试验，最优前处理条件为加入6 mol· L^-1盐酸溶液10 mL，5％乙醇5 mL，105℃水解6 h。结果：线性范围为0.001～0.3 mg· mL^-1（r＝0.9999），平均加样回收率为97.0％，RSD＝0.5％（n＝9）。结论：本方法简便、准确、专属性强，可用于测定鞣酸蛋白原料及制剂中鞣酸的含量。

Objective：To establish an HPLC method for the determination of tannic acid in albumin tannate and its preparations.Methods：The column was Techmate C18-ST （4.6 mm ×250 mm, 5μm）.The mobile phase A was 0.5%acetic acid and the mobile phase B was acetonitrile ,the flow rate was 0.8 mL· min^-1 with gradient elution;The column temperature was 30.0℃and the injection volumn was 10μL with a detection wavelength at 272 nm.Orthogonal test was designed by SPSS 19.0 software and results showed that the optimal pretreatment conditions was to join 10 ml of 6 mol· L^-1 hydrochloric acid solution and 5 ml of 5%ethanol, hydrolyzing for 6 hours at 105℃.Results： The linear range was 0.001-0.3 mg · mL^-1 （ r=0.9999 ） , the average recovery rate was 97.0% and RSD=0.5（n=9）.Conclusion： The method is simple, accurate and suitable for determining the content of tannic acid in albumin tannate and its preparations.