HPLC测定胸腺素α1-胸腺五肽融合肽的含量

Determination of Thymosin α1-Thymopentin Fusion Peptide

目的建立高效液相色谱法(HPLC)测定胸腺素α1-胸腺五肽融合肽含量的方法。方法采用Agilent ZORBAX 80A Extend-C_(18)(250 mm×4.6 mm,5μm)色谱柱,以0.05 mol/L磷酸氢二钠溶液(用磷酸调p H至7.5)-乙腈(9:1,v/v)为流动相,进样量为20μl,检测波长为208 nm,流速为0.6 ml/min;以峰面积归一化法计算含量。结果胸腺素α1-胸腺五肽融合肽在0.479~1.367 mg/ml范围内线性关系良好(r=0.9998),定量限为19.14 ng/ml,检出限为6.38 ng/ml。结论该方法专属性强,重复性和精密度均较好,结果准确,可用于测定胸腺素α1-胸腺五肽融合肽的含量。

Objective To establish an HPLC method for determination of Thymosinα1-Thymopentin Fusion Peptide.Methods The Agilent ZORBAX 80A Extend-C18(250 mm×4.6 mm,5μm)was adopted,using 0.05 mol/L phosphate buffer(pH 7.5)-acetonitrile(9:1,v/v)as mobile phase.The flow rate was 0.6 ml/min,the injection volume was 20μl and the detection wavelength was set at 208 nm.The content were calculated by peak area normalization method.Results The linear range of Thymosinα1-Thymopentin Fusion Peptide was 0.479-1.367 mg/ml(r=0.9998).The limit of quantification was 19.14 ng/ml and the limit of detection was 6.38 ng/ml.Conclusion The HPLC method was proved to be accurate,sensitive and specific,which can be used for the determination of Thymosinα1-Thymopentin Fusion Peptide.