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目的:建立一类新药Ⅱ型胶原蛋白的液相色谱分析方法。方法:选择鸡软骨为提取原料制成的Ⅱ型胶原蛋白,采用分子排阻柱高效液相色谱法测定。色谱条件:使用Sepax nanofilm SEC-150(4.6mm×300mm,5μm)色谱柱,流动相为0.15mol·L-1磷酸钾盐缓冲液(pH6.7),流速0.35mL.min-1,UV检测波长210nm,柱温25℃。结果:Ⅱ型胶原蛋白浓度在0.0894～0.8940mg·mL-1范围内,线性关系良好(r=0.9998);重复性、稳定性良好。结论:该方法简便、准确,可用于测定鸡软骨Ⅱ型胶原蛋白的含量。
Objective: To establish a new type of liquid chromatography of collagen Ⅱ. Methods: The type Ⅱ collagen prepared from chicken cartilage as raw material was selected and determined by size exclusion column (HPLC). Chromatographic conditions: using a Sepax nanofilm SEC-150 (4.6 mm × 300 mm, 5 μm) column with a mobile phase of 0.15 mol·L -1 potassium phosphate buffer (pH 6.7) at a flow rate of 0.35 mL.min -1, UV detection Wavelength 210nm, column temperature 25 ℃. Results: The collagen type Ⅱ in the range of 0.0894 ~ 0.8940 mg · mL-1 showed a good linear relationship (r = 0.9998). The reproducibility and stability were good. Conclusion: The method is simple and accurate and can be used to determine the type Ⅱ collagen content of chicken cartilage.