高效液相色谱-质谱法测定棉籽仁中左旋和右旋棉酚含量

Determination of(-)-and(+)-gossypol in cottonseed by high-performance liquid chromatography with mass spectrum

建立一种高效液相色谱一质谱法测定棉籽仁中（-）-棉酚和（＋）-棉酚含量的检测方法。样品经石油醚脱脂，含10％冰醋酸的N，N-二甲基甲酰胺超声波提取，以（R）-（-）-2-胺基-1-丙醇为手性拆分试剂进行柱前衍生化反应，乙腈：1．0％甲酸=85：15作为流动相，用AgilentXDB—C18色谱柱（250ramx4．6ram，5μm）分离，流速1mL／min，柱后分流比15％，在电喷雾正离子模式下对m／z633．2进行选择性离子监测。该方法下棉酚在1．93-38．6μg/mL范围内线性关系良好，（-）-和（＋）-棉酚在低、中、高的三个添加水平范围内的平均回收率为99．4％-103．0％，RSD值为1-27％～3．86％；（-）-和（＋）-棉酚的检出限分别为2．52、3．41ng／mL，定量限分别为8．39、11．4ng/mL。结果表明，此法操作简便、快速、灵敏、准确，样品处理简便易行，适于测定棉籽仁中（-）和（＋）-棉酚的含量。

A HPLC with mass spectrum derivatization method was developed for the determination of（-）-and（ ＋ ） -gossypol in cottonseed.After degreased by petroleum ether,the samples were extracted with ultrasoninc by 10% glacial acetic acid of N, N- dimethyl formamide formamide and （R） - （ - ） -2- amino- 1 - propanol was selected as the chiral separation reagent.Then the chromatographic separation was carried out on an Agilent XDB-C18 column （250mm x4.dmm,5μm）with a mobile phase consisted of 85% acetonitrile and 15% water within 1.0% formic acid at a flow rate of 1mL/min.The condition of mass spectrometer was operated at m/z 633.2 in positive elctrospray ionization mode.It showed that a good linear relationship between HPLC peak area and gossypol concentration was obtained over the range of 1.93-38.6pg/mL,mean spike recoveries of（-）-and（ ＋ ）-gossypol at three levels were 99.4% - 103.0%, with RSD ranged from 1.27%-3.86%.The limits of detection were 2.52ng/mL and 3.41ng/mL, while the limits of quantification were 8.39ng/mL and l 1.4ng/mL for （-）- and （ ＋ ）- gossypol respectively.The method provided simplicity, rapidity, sensitivity and accuracy, and therefore was applicable to the determination of （ - ） and （ ＋ ） -gossypol in cottonseed.