Li Jiong, Wang Jiaofei, Qiu Hongyu, Li Wei
Hangzhou Institute for Food and Drug Control, Hangzhou 310017, China.
Se Pu. 2017 Aug 8;35(8):832-836. doi: 10.3724/SP.J.1123.2017.04007.
A modified high performance liquid chromatographic (HPLC) method was developed for the determination of the five nucleotides (uridine monophosphate (UMP), adenosine monophosphate (AMP), inosine monophosphate (IMP), guanosine monophosphate (GMP) and cytidine monophosphate (CMP)) in infant formula milk powder. The samples were extracted by water, deproteinized by acetic acid and purified with an HLB SPE cartridge. The analytes were separated by a Waters XBrigde Amide column (150 mm×4.6 mm, 3.5 μ m). Acetonitrile, 10 mmol/L sodium dihydrogen phosphate aqueous solution and 0.12%(v/v) phosphoric acid aqueous solution were used as mobile phases with gradient elution. The detection wavelength of photodiode array detector was set at 254 nm. Five linear calibration curves were obtained with correlation coefficients () of 0.9999. The recoveries were determined at three spiked levels ranging from 86.9% to 105.7%. The limits of quantification (LOQs) were from 5.6 mg/kg to 8.0 mg/kg. The intra-day and inter-day precisions were 0.5%-1.7% (=5) and 0.6%-1.9% (=9), respectively. The method is simple, effective, accurate and repeatable. It is suitable for thedetermination of the five nucleotides in infant formula milk powder.
建立了一种改进的高效液相色谱(HPLC)法,用于测定婴幼儿配方奶粉中的5种核苷酸(尿苷一磷酸(UMP)、腺苷一磷酸(AMP)、肌苷一磷酸(IMP)、鸟苷一磷酸(GMP)和胞苷一磷酸(CMP))。样品用水提取,用乙酸脱蛋白,并用HLB固相萃取柱净化。采用Waters XBrigde酰胺柱(150 mm×4.6 mm,3.5μm)分离分析物。以乙腈、10 mmol/L磷酸二氢钠水溶液和0.12%(v/v)磷酸水溶液为流动相进行梯度洗脱。将光电二极管阵列检测器的检测波长设置为254 nm。获得了5条线性校准曲线,相关系数()为0.9999。在3个加标水平下测定回收率,范围为86.9%至105.7%。定量限(LOQ)为5.6 mg/kg至8.0 mg/kg。日内和日间精密度分别为0.5%-1.7%(=5)和0.6%-1.9%(=9)。该方法简单、有效、准确且可重复。适用于婴幼儿配方奶粉中5种核苷酸的测定。