伤口愈合肽HPLC分析方法的研究

目的：研究高效液相色谱法（HPLC）分析伤口愈合肽的方法。方法：HPLC测定用Kromasil-C18色谱柱（250mm＊4.6mm,5滋m）,拟确定的色谱条件为：二元线性梯度洗脱,流动相A：0.1%三氟乙酸水溶液;流动相B：0.1%三氟乙酸50%乙腈,流速为1ml/min,检测波长为215nm。结果：线性梯度洗脱条件：起始流动相为A70%：B30%,洗脱最终流动相为A30%：B70%;伤口愈合肽浓度在0.1-0.3mg/ml内,其标准曲线的相关系数为R2=0.9998,回归方程为：y=237.19x-0.3249;日内、日间相对标准偏差都不大于2%;重复性的相对标准偏差都小于药典规定的2.0%。结论：该方法稳定性、重复性良好,符合伤口愈合肽新药研究的实验检测要求。

Study on Method of HPLC Analysis of Wound Healing Peptide

Objective： To establish a HPLC method for analysis of wound healing peptide.Methods： The analysis was performed on Kromasil-C18（250mm ＊4.6mm,5μm）column by linear gradient elution with the mobile phase of A ： 0.1% trifluoroacetic acid and B： 0.1% TFA 50% acetonitrile.The flow rate was 1ml/min and the detection wavelength was 215nm.Results： The suitable mobile phase of linear gradient elution is from A70%：B30% to A30%： B70%;The regression equation and correlation coefficient for the standard curve of wound healing in 0.1-0.3mg/ml are y = 237.19x-0.3249 and R2 = 0.9998 respectively.Both relative standard deviations of Intraday,extraday variation and repeatability were less than 2% in pharmacopoeia.Conclusion： The method of HPLC is stable,repeatable and it is suitable for determination of wound healing peptide in new medicine study.