目的 建立定量核磁共振氢谱法测定β-烟酰胺单核苷酸(NMN)含量的方法。方法 通过核磁共振氢谱法测定NMN含量,以对苯二酚为内标,延迟时间为4 s,采样次数为16次,以NMN中δ 8.89的峰为定量峰,对苯二酚中δ 6.67的峰为内标定量峰,根据积分峰面积比与质量比计算NMN的含量。结果 定量核磁共振氢谱法中,NMN与内标对苯二酚的质量比在1.255~7.453内有良好的线性关系,线性相关系数(r2)为0.999 8,NMN的加标平均回收率为98.82%,RSD=1.80%,用高效液相色谱法测得NMN的含量为98.56%,二者的相对误差为1.93%。在复杂基质样品中,定量核磁共振氢谱法所测NMN平均含量为0.67%,RSD为1.88%,与标示量0.7%相近,采用高效液相法无法准确定量。结论 该方法操作简便,检测快速准确,可用于测定复杂基质中NMN的含量。
Abstract
: OBJECTIVE To establish a quantitative nuclear magnetic resonance method and determine the content of nicotinamide mononucleotide (NMN). METHODS Hydroquinone was selected as internal standard. Relaxation delay was 4 s and numbers of scan was 16 times. The peak of δ 8.89 in NMN and peak of δ 6.67 in hydroquinone were used as the quantitative peaks. The content of NMN was calculated according to the integral peak area ratio and mass ratio. RESULTS In the mass ratio range of NMN and hydroquinone between 1.255-7.453, a good linear relationship was established with r2=0.999 8. The average recovery of NMN was 98.82% (RSD=1.80%). The content of NMN was 98.56% determined by HPLC, and the relative error was 1.93%. The average content of NMN in complicate matrix samples determined by qNMR was 0.67% (RSD=1.88%), which was similar to the labeled amount 0.7%. However, HPLC method cannot be used for accurate quantification of complicate matrix samples. CONCLUSION This method is simple, rapid and accurate. It can be used for determination of the content of NMN in complicate matrix samples.
关键词
烟酰胺单核苷酸 /
对苯二酚 /
定量核磁共振法 /
高效液相色谱法
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Key words
nicotinamide mononucleotide /
hydroquinone /
quantitative nuclear magnetic resonance method /
high-performance liquid chromatography
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中图分类号：
R917
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参考文献
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脚注
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基金
国家重点研发计划“中医药现代化研究”重点专项2017年度项目“苦参等大宗中药材的综合利用技术研究”资助(2017YFC1701904)
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