鹿茸、鹿心和鹿血中胰岛素样生长因子-Ⅰ的制备及含量检测

Preparation and Determination of Insulin-like Growth FactorⅠ in Deer Antler,Heart and Blood

目的:对鹿茸中胰岛素样生长因子-Ⅰ(IGF-Ⅰ)的制备方法进行研究,并对鹿茸、鹿心和鹿血中IGF-Ⅰ的含量进行检测和分析比较。方法:采用氨-醋酸铵缓冲液超声提取鹿茸、鹿心和鹿血中的IGF-Ⅰ,并利用超滤法和固相萃取法对IGF-Ⅰ提取液进行浓缩与分离纯化,同时平行采用乙醇沉淀法对鹿茸IGF-Ⅰ超滤液进行纯化,以放射免疫法(RIA)测定鹿不同组织部位中IGF-Ⅰ的含量。结果:鹿茸以固相萃取法所得IGF-Ⅰ仅存在于30%甲醇-水洗脱液中,并且其含量(60.8 ng/g)显著高于乙醇沉淀法所得IGF-Ⅰ(46.1 ng/g);鹿茸、鹿心和鹿血中IGF-Ⅰ含量差异较大,鹿茸粉中为61.9 ng/g,鹿血中为21.9 ng/m L,鹿心中未检出。结论:鹿茸IGF-Ⅰ制备方法中固相萃取法优于乙醇沉淀法;鹿茸中IGF-Ⅰ的含量明显高于鹿血和鹿心,是提取鹿茸IGF-Ⅰ的最佳组织部位。

Objective： To optimize the method for preparation of the insulin-like growth factorⅠ（IGF-Ⅰ） in deer antler,and to determine the IGF-Ⅰin deer antler,heart and blood. Methods： Ultrasonic extraction was used to extract IGF-Ⅰfrom different tissues of deer with ammonia-ammonium acetate buffer,followed by ultrafiltration and solid phase extraction to concentrate and purify the samples. At the same time,ethanol precipitation method was carried out in the purification of IGF-Ⅰultrafiltratein deer antler,a parallel test proceeded and radio immune assay（RIA） was set to determine the IGF-Ⅰin deer antler,heart and blood. Results： The IGF-Ⅰ（60. 8 ng/g） in deer antler by solid phase extraction was only existed in 30% methanol aqueous solution which was much higher than that（46. 1 ng/g）by ethanol precipitation method. The quantities of IGF-Ⅰin deer antler,heart and blood were significantly different,it was 61. 9 ng/g in antler and 21. 9 ng/m L in blood,while there was no IGF-Ⅰtested in deer heart. Conclusion： Solid phase extraction is superior to ethanol precipitation method in preparing IGF-Ⅰ in deer antler and it is clear that the IGF-Ⅰ contained in deer antler is significantly higher than that in deer heart and blood,so it is the best choice to take IGF-Ⅰfrom deer antler.