柱前手性衍生化-高效液相色谱法测定棉籽仁中左旋和右旋棉酚含量

Determination of (-) and (+)-Gossypols in Cottonseed Kernel by High-Performance Liquid Chromatography with Chiral Pre-column Derivatization

目的:建立柱前手性衍生化-高校液相色谱法测定棉籽仁中(-)-棉酚和(+)-棉酚含量的检测方法。方法:采用XDB-C18色谱柱(250mm×4.6mm,5μm),流动相为乙腈-10mmol/L KH2PO4缓冲溶液(磷酸调pH3.0)(80:20,V/V),(R)-(-)-2-胺基-1-丙醇为手性拆分试剂,流速1.0mL/min,柱温40℃,检测波长247nm。结果:(-)-棉酚在1.96～29.46μg/mL质量浓度范围内线性关系良好,回收率为99.7%～104.1%,RSD值为1.50%～3.07%;(+)-棉酚在1.82～27.30μg/mL质量浓度范围内线性关系良好,回收率为100.6%～105.8%,RSD值为1.08%～3.53%。结论:此法操作简便、快速、灵敏、准确,样品处理简便易行,可用于棉籽仁及相关食品中(-)-和(+)-棉酚的含量检测。

An HPLC with chiral pre-column derivatization method was developed for the determination of （--）-and （＋）-gossypols in cottonseed kernel. Chromatographic separation was carried out on an Agilent XDB-C18 column （250 mm × 4.6 mm, 5 μm） with a mobile phase which consisted of 80% acetonitrile and 20% 10 mmol/L KH2PO4 buffer adjusted to pH 3.0 with H3PO4 at a flow rate of 1.0 mL/min. （R）-（--）-2-amino-l-propanol was used as a chiral separation reagent and the detection wavelength was set at 247 nm. A good linear relationship between HPLC peak area and （--）-gossypol concentration was obtained over the range of 1.96 to 29.46 lag/mL, and the mean recoveries of （--）-gossypol at three spiked levels were 99.7% -- 104.1%, with RSDs rangeing from 1.50% to 3.07%. As for （＋）-gossypol, the linear range was 1.82-- 27.30 μg/mL, mean recoveries at three spiked levels were 100.6%- 105.8% with RSDs ranging from 1.08% to 3.53%. The method provides simplicity, rapidity, sensitivity and accuracy, and therefore is applicable to the determination of （-）-and （＋）-gossypols in cottonseed kernel and related foods.