高效液相色谱法测定星虫体壁胶原蛋白

Determination of parietal collagen in Sipunculid by high performance liquid chromatography

目的建立快速检测胶原蛋白的高效液相色谱法。方法采用异硫氰酸苯酯柱前衍生反相高效液相色谱，以乙晴-醋酸盐缓冲系统梯度洗脱，于254nm波长处根据羟脯氨酸的含量来推算星虫体壁胶原蛋白。结果羟脯氨酸衍生物与其它氨基酸较好分离，在5～200μg／mL浓度范围内有良好的线性关系（r=0．9999），回收率为102．6％，RSD为1．58％。结论此法快速、灵敏、分辨率高，适用于动物组织中胶原蛋白含量的测定。

Purpose To set up a method of HPLC to determine the content of parietal collagen in Sipunculid. Methods RP-HPLC was used by precolumn derivation with phenylisothiocyanate. Acetonitrile-acetate buffer linear gradient elution was used. Detecting wavelength was 254 nm. Results Derivation of hydroxyproline could be separated from other amino acids. Hydroxyproline showed good linear（ r = 0. 999 9） in the range of 5 -200 μg/mL. The recovery was 100.1% - 104.6%. RSD was 1.58% .Conclusion The method is rapid, and accurate, and it is suitable for detection of collagen samples obtained from animal tissues.