高效液相色谱法测定牛初乳IGF含量

Determination of bovine colostrum IGF by HPLC

目的： 为定性、定量牛初乳 Ｉ Ｇ Ｆ 提供一种非电泳的分析手段。方法：应用 Ｓｐｈｅｒｉｓｏｒｂ Ｃ１８ 色谱柱和乙腈溶液组成的 Ｈ Ｐ Ｌ Ｃ 色谱系统分析牛初乳 Ｉ Ｇ Ｆ。结果：用此色谱系统从牛初乳 Ｉ Ｇ Ｆ 中分离出多种紫外吸收光谱各不相同的非 Ｉ Ｇ Ｆ 成分。研究表明 Ｈ Ｐ Ｌ Ｃ 法分辨率高于电泳法，分析灵敏度相近。其最小检测浓度为２５０ ｎｇ·（５０ μｌ） － １（ 信噪比＞ ２） 。发现 Ｉ Ｇ Ｆ 是一种不稳定的多肽，其合适的储存条件有待研究。结论： Ｈ Ｐ Ｌ Ｃ 法因有较高的分辨率，可结合电泳法用于需要鉴别 Ｉ Ｇ Ｆ

OBJECTIVE:To develop a non electrophoresis method for qualitative and quantitative analysis of bovine colostrum IGF.METHODS:IGF was analyzed by an HPLC system consisted of sephrisorb C 18 with acetonitrile solution.RESULTS:Several non IGF components were separated from IGF from bovine colostrum.Each of them showed its own characteristic UV absorption spectrum scanned from 190 nm to 360 nm.The results demonstrated that the method of HPLC for resolution of IGF surpassed to that of electrophoresis,the quantitative sensitivity for HPLC was close to that for the latter.Its minimum detect concentration was 250 ng·(50 μl) -1 corresponding to a signal to noise ratio of 2.New finding was that IGF was a labile peptide,further investigation might be necessary for its suitable condition of storage.CONCLUSIONS:HPLC,due to its higher resolution,can be applied,conjugated with electrophoresis,in the areas of purity identification and quantitation of IGF.