HPLC法同时测定核苷酸花粉胶囊中5种核苷酸含量

目的:建立HPLC法以测定核苷酸花粉胶囊中的5种核苷酸(胞苷酸二钠、尿苷酸二钠、鸟苷酸二钠、肌苷酸二钠、腺苷酸)。方法:以流动相作为提取溶剂对样品进行超声提取;选用Waters Atlantis T3(4.6 mm×250 mm,5 μm)色谱柱,以10 mmol·L^-1磷酸二氢钾缓冲液(以磷酸调pH 3.9)为流动相,检测波长260 nm。结果:样品中5种核苷酸成分得到很好的分离,5种核苷酸线性良好,相关系数0.999 4~0.999 7;平均回收率(n=6)为88.25%~101.3%,RSD为0.94%~4.04%;3批样品中核苷酸总量分别是293、287、289 mg·g^-1。结论:本方法操作简单快速、灵敏度高、重现性好,准确可靠,能够满足日常分析检测的需要,为核苷酸类保健食品的质量控制提供了简便方法。

Objective:To establish an HPLC method for the determination of five kinds of nucleotides [including cytidine 5'-monophosphate disodium salt(5'-CMPNa₂),uridylic-5'-monophosphate disodium salt(5'-UMPNa₂),guanosine-5'-monophosphate disodium salt(5'-GMPNa₂),inosine 5'-monophosphate disodium salt(5'-IMPNa₂),5'-adenylic acid] in nucleotide pollen capsules.Methods:The chromatographic separation was achieved on a Waters Atlantis T3 column(4.6 mm×250 mm,5 μm),using 10 mmol·L^-1 KH₂PO₄ buffer solution(adjusted to pH 3.9 with phosphoric acid)as the mobile phase at a flow rate of 1.0 mL·min^-1,and the detection wavelength was set at 260 nm.Results:Five nucleotides were well separated within 25 minutes.The five calibration curves of nucleotides showed good linearity with correlation coefficient of 0.999 4-0.999 7.The average recovery ranges(n=6)were between 88.25% and 101.3% with RSDs of 0.94%-4.04%.In the 3 batches of samples,the total contents of nucleotides were 293 mg·g^-1,287 mg·g^-1 and 289 mg·g^-1.Conclusion:The method is simple,rapid,sensitive,reproducible,accurate and reliable,which can meet the needs of daily analytical testing,and can be applied to the quality control of five nucleotides in health products.