高效液相色谱法测定乳粉中的核苷酸

HPLC method for determination of nucleotides in infant formula milk powder

为检测乳粉中胞嘧啶核苷酸、尿嘧啶核苷酸、腺嘌呤核苷酸、鸟嘌呤核苷酸和次黄嘌呤核苷酸5种核苷酸的质量浓度,建立了乳粉中5种核苷酸的紫外检测液相色谱法。试样经质量分数为10%乙酸去除蛋白质,SUPELCOSIL LC-18-T C18柱分离,流动相为1.40mmol/L四丁基硫酸氢铵磷酸盐缓冲液,甲醇体积分数5%,254 nm紫外检测器液相色谱分析方法定量检测。胞嘧啶核苷酸、腺嘌呤核苷酸、尿嘧啶核苷酸、鸟嘌呤核苷酸和次黄嘌呤核苷酸的平均回收率分别是93.2%,103.4%,106.2%,94.8%和103.7%。CMP,AMP,UMP,GMP和IMP10次样品重复测定变异系数分别为1.4%,1.6%,1.7%,1.8%和1.7%。

For the determination of 5 nucleotides in infant formula milk powder, the HPLC UV absorbance detector method was established. We established the method in which 10% acetic acid was used for elimination of protein, SUPELCOSIL LC-18-T column for isolation and the nucleotides were determined quantitatively at UV254 nm wavelength with HPLC. The mobile phase was phosphate and methanol. The average recovery rates of cytidine, uridine, adenosine, guanosine and inosine were 93.2%, 103.4%, 106.2%,94.8%, 103.7%,respectively and the coefficient variance were 1.4%, 1.6%, 1.7%, 1.8% and1.7%.