超高效液相色谱法测定婴幼儿配方奶粉中5种核苷酸

Determination of five nucleotides in infant formula milk powder by Ultra Performance Liquid Chromatography

建立并优化了超高效液相色谱检测婴幼儿配方奶粉中5种核苷酸(尿苷酸(UMP)、腺苷酸(AMP)、肌苷酸(IMP)、鸟苷酸(GMP)、胞苷酸(CMP))的方法。样品用水提取后,经乙酸沉淀蛋白质和Oasis HLB固相萃取柱净化,采用Waters BEH Amide(1.7μm,2.1×100 mm)色谱柱分离,以乙腈、10 mmol/L磷酸二氢钠溶液和0.1%(v/v)磷酸水溶液为流动相进行梯度洗脱,紫外检测器(波长为254 nm)检测。标准曲线相关系数为0.9995~0.9999;回收率为97.0%~102%之间;6次样品测定5种核苷酸RSD为2.5~4.7%;定量限为1.5~2.0 mg/kg。该方法分离效果好,回收率高,重复性好,可作为婴幼儿配方奶粉中5种核苷酸的有效检测方法。

A method for the determination of five nucleotides(UMP, AMP, IMP, GMP and CMP) in infant formula milk powder by ultra performance liquid chromatography(UPLC) was established and optimized. The samples were extracted by water, deproteinizde by acetic acid and purified with an Oasis HLB SPE cartridge. The analytes were separated by a waters BEH amide column(1.7 μm,2.1×100 mm),acetonitrile、10 mmol/L sodium dihydrogen phosphate aqueous solution and 0.1%(v/v)phosphoric acidaqueous solution were used as mobile phases with gradient elution.The detection wavelength UV detector was set at 254 nm. The relative coefficient was 0.9995~0.9999, the recovery97.0%~102%;the RSD in 6 samples by testing was 2.5%~4.7%. The limits of quantification were from 1.5 mg/kg to 2.0 mg/kg. The method is effective,accurate and repeatable..It is suitable for the determination of the five nucleotides in infant formula milk powder.