Abstract:

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-1-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 μg/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.

Key words: cottonseed, (－)-gossypol, (＋)-gossypol, (R)-(－)-2-amino-1-propano, chiral pre-column derivatization, high performance liquid chromatography (HPLC)