<i>β</i>-烟酰胺单核苷酸跨境产品中NMN含量的测定

张文宇; 兰韬; 赵溪; 吴琦; 初侨; 于聪聪; 王大宏; 张维冰; 云振宇

doi:10.13386/j.issn1002-0306.2021110015

β-烟酰胺单核苷酸跨境产品中NMN含量的测定

doi: 10.13386/j.issn1002-0306.2021110015

张文宇^{1, 2,},
兰韬^2, ,,
赵溪³,
吴琦²,
初侨²,
于聪聪²,
王大宏⁴,
张维冰^{1, 5},
云振宇^2, ,

1.
齐齐哈尔大学，黑龙江齐齐哈尔 161006
2.
中国标准化研究院，北京 100191
3.
汤臣倍健股份有限公司，广东珠海 519040
4.
庶正康讯（北京）商务咨询有限公司，北京 100054
5.
华东理工大学，上海 200237

基金项目: 中央级公益性科研院所基本科研业务费（562020Y-7483）；国家市场监督管理总局科技计划项目（2020YJ044）。

详细信息

作者简介:
张文宇（1998−），男，硕士研究生，研究方向：分析化学，E-mail：1042755540@qq.com

通讯作者:
兰韬（1984−），男，副研究员，研究方向：保健食品标准化，E-mail：lantao@cnis.ac.cn

云振宇（1978−），男，博士，研究员，研究方向：保健食品标准化，E-mail：yunzy@cnis.ac.cn

中图分类号: TS207.3
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- 被引次数: 0
出版历程
- 收稿日期: 2021-11-03
- 网络出版日期: 2022-05-10
- 刊出日期: 2022-05-15

Determination of β-Nicotinamide Mononucleotide (NMN) in NMN Cross Border Products

ZHANG Wenyu^{1, 2
,},
LAN Tao^{2
, ,},
ZHAO Xi³,
WU Qi²,
CHU Qiao²,
YU Congcong²,
WANG Dahong⁴,
ZHANG Weibing^{1, 5},
YUN Zhenyu^{2
, ,}

1.
Qiqihar University, Qiqihar 161006, China
2.
National Institutes for Food and Drug Control, Beijing 100191, China
3.
Byhealth Co., Ltd., Zhuhai 519040, China
4.
Shuzheng Kangxun (Beijing) Business Consulting Co., Ltd., Beijing 100054, China
5.
East China University of Science and Technology, Shanghai 200237, China

摘要

摘要: 本文基于高效液相色谱法，通过优化色谱条件和样品前处理条件，建立了一种测定β-烟酰胺单核苷酸（NMN）胶囊和NMN片剂基质中NMN含量的检测方法，并进行方法学验证。结果表明，本方法在5~500 μg/mL范围内有较好的线性关系，方法检出限（LOD，S/N=3）为1.0 mg/kg，定量限（LOQ，S/N=10）为3.0 mg/kg，仪器精密度为0.3%。对于NMN片剂样品，方法稳定性为1.7%，重复性为1.8%；对于NMN胶囊样品，方法稳定性为0.6%，重复性RSD值0.6%。NMN片剂和NMN胶囊样品中NMN的检测回收率在90.9%~108.9%之间，相对标准偏差均不超过1.9%。综上，本方法具有较好的重现性、精密度、稳定性、重复性，结果准确可靠，可用于实际样品中NMN含量的准确测定。通过对6种市售的NMN产品进行检测，所有样品中均检出了NMN，但是部分产品的实际含量较其标称含量偏少，发现部分产品存在虚报NMN含量的问题。本文研究为制定NMN跨境产品中NMN含量的检测方法标准奠定了方法学基础，对于提高NMN相关产品质量，促进相关产业的便捷监管具有深远意义。
- 膳食补充剂 /
- β-烟酰胺单核苷酸 /
- 高效液相色谱法
Abstract: By optimizing the chromatographic conditions and sample pretreatment conditions, a method for determination of NMN content in NMN capsules and NMN tablets matrix was established based on high performance liquid chromatography, and the methodology was verified. The results showed that, the method with range of 5~500 μg/mL had a good linear relationship. The limit of detection (LOD, S/N=3) of NMN was 1.0 mg/kg, while the limit of quantitative (LOQ, S/N=10) was 3.0 mg/kg. The precision of the instrument was 0.3%. For NMN tablet samples, the stability of the method was 1.7% and the repeatability was 1.8%, while for NMN capsule samples, the stability of the method was 0.6%, and the repeatability RSD value was 0.6%. The recovery of NMN in NMN tablets and NMN capsules was between 90.9% and 108.9% with the RSD all no more than 1.9%. In conclusion, this method had good reproducibility, precision, stability and repeatability, could give accurate and reliable results, and could be used for the accurate determination of NMN in actual samples. Through the detection of 6 kinds of commercially available NMN products, NMN was detected in all samples, but the actual contents of some products were less than their nominal content. It was found that some products had the problem of false reporting of NMN content. This study would lay a methodological foundation for formulating the detection method standard of NMN content in NMN related dietary supplement products, and was of far-reaching significance for improving the quality of NMN related products and promote the convenient supervision of related industries.
- dietary supplement /
- β-nicotinamide mononucleotide (NMN) /
- HPLC

HTML全文

图 1 β-烟酰胺核苷酸的分子结构式

Figure 1. Molecular structure formula of β-nicotinamide nucleotide

下载: 全尺寸图片幻灯片

图 2 NMN片剂样品基质（a）和NMN胶囊样品基质（b）的流动相优化分离色谱图

Figure 2. Chromatograms of NMN tablet sample matrix (a) and NMN capsule sample matrix (b) based on mobile phase optimization

下载: 全尺寸图片幻灯片

图 3 甲酸加入前后NMN片剂（a）NMN胶囊剂（b）样品基质分离谱图

Figure 3. Chromatograms of NMN tablets matrix (a) and NMN capsule sample matrix (b) before and after formic acid addition

下载: 全尺寸图片幻灯片

表 1 不同溶剂对NMN提取效率的影响（n=3）

Table 1. Effects of different solvents on the extraction efficiency of NMN (n=3)

标准品	水溶液		85%甲醇水溶液		50%甲醇水溶液
标准品	回收率（%）	RSD（%）	回收率（%）	RSD（%）	回收率（%）	RSD（%）
NMN	70.5	1.1	65.3	7.6	95.7	0.4

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表 2 NMN的线性范围、LOD和LOQ

Table 2. Standard curve, linear range, LOD and LOQ of NMN

标准品	保留时间（min）	线性方程	r	线性范围（μg/mL）	LOD（mg/kg）	LOQ（mg/kg）
NMN	5.53	Y=6.76X−13.44	0.999	5.0~500.0	1.0	3.0

下载: 导出CSV

表 3 精密度实验结果（μg/mL）

Table 3. Precision test results (μg/mL)

标准品	1	2	3	4	5	6	RSD（%）
NMN	86.4	86.1	86.3	86.1	86.9	86.4	0.3

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表 4 稳定性实验结果（mg/kg）

Table 4. Results of stability experiments (mg/kg)

基质	1	2	3	4	5	6	RSD （%）
NMN（片剂）	40.7	40.9	41.8	41.9	41.8	40.3	1.7
NMN（胶囊）	141.0	142.3	142.0	142.4	142.9	143.4	0.6

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表 5 重复性实验结果（mg/kg）

Table 5. Results of repeatability experiment (mg/kg)

基质	1	2	3	4	5	6	RSD （%）
片剂	190.7	193.8	193.3	199.7	190.1	191.4	1.8
胶囊	264.4	262.3	262.7	263.3	260.2	263.7	0.6

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表 6 加标回收率实验结果

Table 6. Experimental results of standard recovery

样品	本底值（mg/kg）	5 mg/kg		10 mg/kg		50 mg/kg
样品	本底值（mg/kg）	回收率（%）	RSD （%）	回收率（%）	RSD （%）	回收率（%）	RSD （%）
NMN片剂	31.8	95.7	0.8	95.7	0.6	108.9	0.2
NMN胶囊	41.2	90.9	0.2	93.1	0.2	94.4	1.9

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表 7 6种不同NMN产品中NMN含量（g/kg，n=3）

Table 7. NMN content in 6 different NMN products (g/kg, n=3)

序号	NMN（测定值）	产品标称
1	65.3	75.0
2	268.0	520.3
3	68.2	188.3
4	486.0	500.0
5	197.0	未标称
6	493.0	500.0

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