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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。可用于棉籽蛋白工业化生产过程中棉酚的快速常规分析测定及其质量监控
The authors developed a method for the determination of free and total gossypol in cottonseed protein by HPLC. Samples were extracted with a mixed solvent (95% ethanol: water: ether: glacial acetic acid, 71.5: 28.5: 20: 0.2, v / v) for 30 min at room temperature and filtered. The free gossypol was separated and separated by μBondapak C18 column, mobile phase (methanol: water: acetonitrile: phosphoric acid, 80:15:5:0.2, v / v) flow rate: 1.0ml / min and UV- (FGP) at 0 for total gossypol (TGP). lmol / L oxalic acid solution (butanone: water, 55.3: 5, v / v as solvent), hydrolyzed at 78 ℃ for 6h, filtered, and then measured in the same way as FGP. Under these conditions, gossypol can be better separated from other kinds of gossypol pigments. The recovery of the HPLC quantitative determination was 98.98% (n = 6) with a coefficient of variation of 3.26%. The experimental results show that this method has the advantages of less sample amount, simple and rapid operation, strong specificity, high sensitivity, good reproducibility and accurate and reliable results. Can be used for rapid routine analysis and quality control of gossypol in industrialized cottonseed protein production