胶囊中β-烟酰胺单核苷酸和烟酰胺腺嘌呤二核苷酸的测定

Determination ofβ-Nicotinamide Mononucleotide and Nicotinamide Adenine Dinucleotide in capsule

建立高效液相色谱法同时测定胶囊中β-烟酰胺单核苷酸(β-nicotinamide mononucleotide,NMN)和烟酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide,NAD+)含量的方法。样品经磷酸盐缓冲溶液(PBS)提取,采用ZORBAX SB-Aq(粒径5μm,4.6 mm×250 mm)进行分离,采用0.1%甲酸水和甲醇作为流动相进行梯度洗脱,紫外检测器测定,外标法定量。在最优条件下,NMN和NAD+在1.00~100 mg/L范围内线性关系良好,相关系数分别为0.9999和0.9999。NMN和NAD+的检出限均为0.1 g/kg,定量限均为0.3 g/kg,在0.3,0.6和3.0 g/kg的加标水平下,NMN和NAD+的回收率为81.6%~101%,相对标准偏差为0.737%~2.15%。该方法灵敏度高、选择性好、准确可靠、操作简单,可用于胶囊中NMN和NAD+含量的同时快速检测。

A method for simultaneous determination ofβ-nicotinamide mononucleotide(NMN)and nicotinamide adenine dinucleotide(NAD+)in capsule by HPLC(high performance liquid chromatography)was established.The sample was extracted with Phosphate buffer(PBS),and ZORBAX SB-Aq chromatographic column was used for gradient elution with 0.1%(V/V)formic acid and methanol and water as mobile phase.It was detected by ultraviolet detector and quantified by external standard.Under the optimized condition,NMN and NAD+had a good linear relationship in the concentration range of 1.00-100 mg/L,and the correlation coefficients of them were 0.9999 and 0.9999,respectively.The limits of detection of NMN and NAD+were 0.1 g/kg,while the limits of quantitation were 0.3 g/kg.The recoveries of NMN and NAD+ranged from 81.6%to 101%at the concentration level of 0.3,0.6 and 3.0 g/kg,and the relative standard deviations were 0.737%-2.15%.The method had the advantages of high sensitivity,good selectivity,accurate and reliable results and simplicity of operator,which was suitable for the simultaneous quantitative analysis of NMN and NAD+in capsule.