高效液相色谱法测定饲料添加剂D-生物素的含量

Determination of D-biotin in feed additives by HPLC

为测定饲料添加剂原料中D-生物素含量,建立并优化了高效液相色谱法(HPLC),采用色谱柱为Agilent XDB-C_(18)反相柱(150 mm×4.6 mm,5μm),流动相为乙腈+缓冲液(含0.1%高氯酸钠和0.1%磷酸的水溶液)=8.5+91.5(体积比),进样量为20μL,检测波长为210 nm,流速为1.2 mL/min。D-生物素在25～400μg/mL范围内线性关系良好,相关系数为1.00000,检出限1.5 mg/kg,方法回收率为99.5%,RSD为0.5%(n=9)。本方法快速简便,能够准确测定饲料添加剂原料中D-生物素的含量,相对于酸碱滴定法有更好的专属性。

To establish and optimize the HPLC method for the determination of D-biotin content in feed additives.Agilent XDB-C 18 （150 minx4.6 mm, 5 p～m） was used as the column,with the detection wavelength set at 210 nm, the mobile phase was acetonitrile and buffer solution （8.5：91.5） （the water solution containing 0.1% sodium perchlorate and 0.1% phosphoric acid）,with the flow rate at 1.2 mL/min,the injection volume was 20 μL.The linear range was 25 ～ 400 μg/mL（r=1.0000） and the detection limit was 1.5 mg/kg.The method recovery rear and RSD was 99.5% and 0.5% （n=9）.This method was not only fast and simple,but also could accurately measure the content of D- biotin in feed additives,and has better specificity than neutralization titration.