Determination of gossypol in feeds by HPLC
2008
Aoyama, K.(Food and Agricultural Materials Inspection Center, Kobe (Japan). Kobe Regional Center)
An HPLC method for determination of gossypol in feed was developed. Gossypol in food was extracted with acetic acid-water-phosphoric acid (85:15:1) for 20 min in a water bath at 100degC. The extract was diluted with acetone-water (1:1), and injected into the HPLC. HPLC was performed with a Shodex C18M4E (4.6 mm i.d. x 250 mm) column at a flow-rate of 1.0 mL/min, using a mobile phase of methanol-water (9:1) adjusted to pH 2.6 with phosphoric acid, and gossypol was detected with a UV detector (254 nm). A recovery test was conducted with cottonseed spiked with gossypol at 1,000 and 5,000 mg/kg, and with 2 kinds of formula feed spiked with gossypol at 58 and 580 mg/kg. The mean recoveries of gossypol were 90.8-105.0% and the relative standard deviations (RSD) were within 3.0%. A collaborative study was conducted with cottonseed and formula feed spiked with gossypol at 305 mg/kg in 8 laboratories. The average content of gossypol in cottonseed was 6,090 mg/kg, and the repeatability and reproducibility as the relative standard deviation (RSDsub(r) and RSDsub(R)) were 3.3% and 4.4%, while the mean recovery, RSDsub(r) and RSDsub(R) of gossypol in formula feed were 87.0%, 2.7% and 5.5%, respectively.
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