HPLC法测定十全大补膏滋剂中氨基酸成分的含量

Determination the Content of Amino Acid in Shiquan Dabu Herb Paste by HPLC

目的:采用HPLC法分析十全大补膏滋剂中氨基酸成分的含量。方法:十全大补膏滋剂加盐酸溶液于150℃水解1h,以异硫氰酸苯酯柱前衍生,HPLC法测定氨基酸含量。采用Agilent Eclipse XDB-C_(18)(250mm×4.6mm,5μm)色谱柱,以乙腈-0.1mol/L醋酸钠溶液(7∶93)为流动相A,以乙腈-水(4∶1)为流动相B进行梯度洗脱,体积流量1.0mL/min,柱温43℃,检测波长254nm。结果:L-羟脯氨酸、甘氨酸、丙氨酸、L-脯氨酸4种氨基酸成分分别在0.137 6~2.752、0.147 6~2.952、0.144 6~2.892、0.130 4~2.608μg内呈良好的线性关系(R^2≥0.999 0),平均加样回收率分别为96.6%、98.2%、96.3%、97.7%,RSD<3.00%。结论:该方法简便、准确,重复性好,可用于十全大补膏滋剂的质量控制。

Objective：To established a HPLC method for determination of amino acids in Shiquan dabu herb paste. Methods： Shiquan dabu herb paste was hydrolyzed for lh at 150℃ using hydrochloric acid, and the amino acids were determined by HPLC af- ter being derived with phenyl isothiocyanate. HPLC was performed on an Agilent Eclipse XDB-C18 （4. 6 × 250mm, 5μm）column. With acetonitrile-sodium acetate solution （7 ： 93） as mobile phase A and acetonitrile-water as mobile phase B in the gradient mode at a flow rate of 1.0mL/min. The column temperature was 43℃, and the detection wavelength was set at 254nm. Results： All the ac- tive constituents calibration curves showed a good linearity, The liner range of L-hydroxyproline, glycine, alanine, L-proline were 0. 137 6~2. 752,0. 147 6~2. 952,0. 144 6~2. 892,0. 130 4~2. 608μg, R2≥0. 999 0, respectively. And the average recovery was 96.6 %, 98.2 %, 96.3 %, 97.7 %, RSD〈3.00%. Conclusion ： The method is simple, accurate, and repeatable, which could be a- vailable for the quality control of Shiquan dabu herb paste.