HPLC法测定阿胶中6种氨基酸的含量

Determination of 6 kinds of amino acid content in Ejiao by HPLC method

目的：通过柱前衍生化,高效液相色谱（HPLC）法测定阿胶中6种氨基酸的含量。方法色谱柱为Hypersil C18（4.6 mm＆#215;250 mm,5μm）；流动相A：乙腈-0.1mol/L乙酸钠（7：93）,用乙酸调节pH为6.5,流动相B：乙腈-水（4：1）,梯度洗脱；柱温为42℃；流速为1.0 ml/min；检测波长254 nm；异硫氰酸苯酯（PITC）柱前衍生化。结果30 min内6种氨基酸分离完全,峰面积与浓度的线性关系良好, r=0.999。加样回收率97.33%~100.17%, RSD〈3。结论 HPLC法测定阿胶中6种氨基酸的含量方法操作流程简单、快速、准确,可用于该制剂的质量控制和检验。

Objective To determine content of 6 kinds of amino acids in Ejiao by HPLC method. Methods The column was Hypersil C18 （4.6 mm x 250 mm, 5μm）; the A of the mobile phase： Acetonitrile -0.1mol/L sodium acetate （7：93）, the PH adjusted to 6.5 with acetic acid, the B of the mobile phase： Acetonitrile water （4：1）, Gradient elution; the column temperature was 42℃; the flow rate was 1 ml/min; the detection wavelength was 254 nm; Phenyl was othiocyanate （PITC） pre column derivatization.Results 6 kinds of amino acids separation completely within 30 minutes; good linear relationship of peak area and concentration, R=0.999, the recovery rate of 97.33%~100.17%, RSD〈3.Conclusion The HPLC method to determine the content of 6 kinds of amino acids of donkey hide gelatin method of operation is simple, fast, accurate and can be used for quality control and inspection of the preparation.