Aoyama Keisuke
Food and Agricultural Materials Inspection Center, Kobe Regional Center, Hyogo, Japan.
Shokuhin Eiseigaku Zasshi. 2008 Aug;49(4):303-7. doi: 10.3358/shokueishi.49.303.
An HPLC method for determination of goosypol 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 100 degrees C. 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.x250 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 (RSD(r) and RSD(R)) were 3.3% and 4.4%, while the mean recovery, RSD(r) and RSD(R) of gossypol in formula feed were 87.0%, 2.7% and 5.5%, respectively.
建立了一种测定饲料中棉酚的高效液相色谱法。食品中的棉酚用乙酸 - 水 - 磷酸（85 : 15 : 1）在100℃水浴中提取20分钟。提取物用丙酮 - 水（1 : 1）稀释后注入高效液相色谱仪。高效液相色谱分析采用Shodex C18M4E（4.6 mm内径×250 mm）色谱柱，流速为1.0 mL/min，流动相为甲醇 - 水（9 : 1），用磷酸调至pH 2.6，棉酚用紫外检测器（254 nm）检测。对添加了1000和5000 mg/kg棉酚的棉籽以及添加了58和580 mg/kg棉酚的两种配方饲料进行了回收率试验。棉酚的平均回收率为90.8 - 105.0%，相对标准偏差（RSD）在3.0%以内。在8个实验室对添加了305 mg/kg棉酚的棉籽和配方饲料进行了协同研究。棉籽中棉酚的平均含量为6090 mg/kg，重复性和再现性以相对标准偏差（RSD(r)和RSD(R)）表示分别为3.3%和4.4%，而配方饲料中棉酚的平均回收率、RSD(r)和RSD(R)分别为87.0%、2.7%和5.5%。
