用高效液相色谱法（HPLC）测定棉籽蛋白中的游离和总棉酚  

DETERMINATION OF FREE AND TOTAL GOSSYPOL IN COTTONSEED PRoTEIN BY HPLC

作　　者：张延坤[1] 房玉水 邓峰[1] 郭大发[1] 

机构地区：[1]天津卫生学环境医学研究所

出　　处：《营养学报》1995年第4期419-424,共6页Acta Nutrimenta Sinica

基　　金：军事医事科学院基金

摘　　要：作者研究建立了一种棉籽蛋白中游离和总棉酚的ＨＰＬＣ测定方法。样品用混合溶剂（９５％乙醇：水：乙醚：冰乙酸，７１．５：２８．５：２０：０．２，ｖ／ｖ）室温　萃取３０ｍｉｎ后，过滤。用μＢｏｎｄａｐａｋＣ１８色谱柱、流动相（甲醇：水：乙腈：磷酸，８０：ｌ５：５：０．２，ｖ／ｖ）流速：１．０ｍｌ／ｍｉｎ、ＵＶ－２５４ｎｍ检测等色谱条件分离测定游离棉酚（ＦＧＰ），对于总棉酚（ＴＧＰ），在０．ｌｍｏｌ／Ｌ草酸溶液中（丁酮：水，５５．３：５，ｖ／ｖ作溶剂），于７８℃水解６ｈ，过滤，再与ＦＧＰ同样方法测定。在此条件下棉酚能与其它类棉酚色素较好地分离。ＨＰＬＣ定量测定的回收率为９８．９８％（ｎ＝６），变异系数３．２６％。实验结果表明，本方法具有样品用量少、操作简便、迅速、专属性强、灵敏度高、重现性好、结果准确可靠的特点。method for the determination of free gossypol (FGP)and total gos-sypol(TGP) in cottonseed protein by HPLC was studied,FGP was extrac-ted with mixed solvents (95% ethanol-water-ether-glacial acctic acid,71.5:28.5:20:0.2,v/v),and detemined on a μBondapak C18 columm with themobile phase (methanol-water-acetonitrile-phosphoric acid,80:15:0.2,v/v)and UV-254nm detector. Under this condition,FGP could be fully separa-ted from gossypol-like pigments. After the sample had been hydrolyzed ina solution of 0.1mol/L oxalic acid(dissolved in 2-butanone-water, 55.3:5,v/v) at 78℃ for 6h and filtered.TGP was determined in the same way asFGP.The quantitative analysis of the gossypol was carried out simultaneous-ly, The recovery rate of the method was 98.98%(n=6) with CV 3.26%.The method was simple,sensitive,rapid and reproducible.

关 键 词：高效液相色谱法 游离棉酚 总棉酚 棉籽蛋白 

分 类 号：R151.3[医药卫生—营养与食品卫生学]