饲料中生物素的高效液相色谱测定

Determination of Biotin in Fodder by HPLC

介绍了用液相色谱测定饲料中生物素的方法,探讨了流动相中缓冲液、pH值、有机溶剂等对分离的影响,确定的较佳色谱条件:HypersilODS柱,流动相为甲醇-0.1mol·L-1KH2PO4(H3PO4调pH＝3.5)(体积比25∶75),流速1.0mL／min,紫外210nm检测;方法的相对标准偏差在3.5%以内,回收率在92%～102%,检出限为1mg／kg;实验表明该法简便、快速,适应性好;方法应用于饲料样品中生物素的测定,取得了很好的结果。

This paper reports the determination of biotin in fodders by HPLC,particularly focusing on the dicusˉsion of the effects of mobile phase on the separation efficiency.It is found that the biotin's peak on chromatogram was tailed and became wider in the system containing only methanol-water without buffer.However,adding KH 2 PO 4 -H 3 PO 4 buffer in mobile phase may effectively improve the chromatogram.By experiment,the optimum chromatographic conditions were identified on Hypersil ODS column(4.6mm×250mm,5μm),with methanol-0.05mol·L -1 KH 2 PO 4 -H 3 PO 4 buffer(25∶75by volume)as a mobile phase,pH value of3.5,flow rate of1.0mL/min,and detected wavelength of UV210nm.The RSD is less than3.5%and the recovery is in the range of92%～102%.The detection limit is1mg/kg.The method has been successfully applied to the determiation of biotin in fodders,which proved to be simple,rapid,and good suitability.