Billedeau S M
Food and Drug Administration, National Center for Toxicological Research, Jefferson, AR 72079.
J Chromatogr. 1989 Jun 23;472(2):371-9. doi: 10.1016/s0021-9673(00)94136-0.
A high-performance liquid chromatographic (HPLC) method for analysis of vitamin K3 (menadione), as the water-soluble sodium bisulfite salt, in synthetic animal feed is described. The menadione sodium bisulfite (MSB) is extracted with aqueous methanol, converted to oil-soluble menadione with sodium carbonate, and partitioned into n-pentane. After evaporation, the menadione is dissolved in methanol and analyzed by reversed-phase HPLC. The menadione is reduced on-line to its fluorescent 1,4-dihydroxy analogue by zinc reduction prior to fluorescence determination at 325 nm excitation wavelength and 425 nm emission wavelength. The fluorescence response was linear in the range 1 to 100 ng of menadione injected. Recovery experiments were performed on synthetic animal feed spiked at 20 and 200 mg/kg levels of MSB. Average recoveries from feed were greater than 90% with an average relative standard deviation of 5.5%. Additional confirmation of menadione in actual feed extracts was performed using capillary gas chromatography-Fourier transform infrared spectroscopy (GC-FT-IR). The HPLC-fluorescence and GC-FT-IR methods had lower limits of detection of 20 micrograms/kg and 2 mg/kg, respectively.
本文描述了一种用于分析合成动物饲料中维生素K3（甲萘醌，以水溶性亚硫酸氢盐形式存在）的高效液相色谱（HPLC）方法。甲萘醌亚硫酸氢钠（MSB）用水性甲醇萃取，用碳酸钠转化为油溶性甲萘醌，再分配到正戊烷中。蒸发后，甲萘醌溶解在甲醇中，通过反相HPLC进行分析。在325nm激发波长和425nm发射波长下进行荧光测定之前，甲萘醌通过锌还原在线还原为其荧光性1,4 - 二羟基类似物。注入的甲萘醌在1至100ng范围内，荧光响应呈线性。对添加了20和200mg/kg水平MSB的合成动物饲料进行了回收率实验。饲料的平均回收率大于90%，平均相对标准偏差为5.5%。使用毛细管气相色谱 - 傅里叶变换红外光谱（GC - FT - IR）对实际饲料提取物中的甲萘醌进行了额外确认。HPLC - 荧光法和GC - FT - IR法的检测下限分别为20微克/千克和2毫克/千克。
