高效液相色谱法测定保健食品中的烟酰胺单核苷酸和烟酰胺腺嘌呤双核苷酸含量

Determination of nicotinamide mononucleotide and nicotinamide adenine dinucleotide in health food by high performance liquid chromatography

中文关键词:  烟酰胺单核苷酸  烟酰胺腺嘌呤双核苷酸  保健食品  液相色谱法

英文关键词:nicotinamide mononucleotide  nicotinamide adenine dinucleotide  health food  high performance liquid chromatography

中文摘要:

      目的 建立高效液相色谱法精确定量保健食品中烟酰胺单核苷酸(nicotinamide mononucleotide, NMN)与烟酰胺腺嘌呤双核苷酸(nicotinamide adenine dinucleotide, NAD?)的含量的分析方法。方法 采用Waters BEH-Amide色谱柱, 0.1%磷酸溶液和乙腈为流动相, 进行梯度洗脱, 流速设定为1.0 mL/min, 进样量为2 μL, 柱温箱温度设置为35 ℃, 选择263 nm作为检测波长。保健食品用60%甲醇溶液超声提取10 min, 将提取液通过微孔滤膜过滤后进行高效液相色谱法测定, 外标法定量。结果 在质量浓度10~1000 μg/mL的范围内, NMN和NAD+均展现出优异的线性关系, 其线性相关系数(r)均不低于0.999, 该方法的定量限为0.50 g/kg, NMN和NAD+的回收率在95.9%~98.8%范围内, 相对标准偏差在1.19%~3.05%之间。结论 该方法可以准确测定保健食品中NMN和NAD+含量。

英文摘要:

      Objective To establish an analytical mehod for the determination of nicotinamide mononucleotide (NMN) and nicotinamide adenine dinucleotide (NAD+) in health food by high performance liquid chromatography. Methods Determination was performed on Waters BEH-Amide column with mobile phase consisting of 0.1% phosphoric acid solution-acetonitrile (gradient elution). The flow rate was 1.0 mL/min, and the sample size was 2 μL. The column temperature was 35 ℃, and the detection wavelength was set at 263 nm. Health food was extracted with 60% methanol solution in ultrasonic water bath for 10 min. The extract was filtered through micoron filter, and analyzed by high performance liquid chromatography. Results The results showed that NMN and NAD+ exhibited excellent linear relationships, the correlation coefficient (r) was over 0.999 in the range of 10–1000 μg/mL. The method limit of quantitation was 0.50 g/kg. Recoveries were ranged from 95.9% to 98.8%, and relative standard deviations were ranged from 1.19% to 3.05%. Conclusion This method is accurate and suitable to determine NMN and NAD+ in health food.