高效毛细管电泳法测定金针菇提取物中β-烟酰胺单核苷酸含量的初步研究

doi:10.13304/j.nykjdb.2021.0728

中国农业科技导报 ›› 2023, Vol. 25 ›› Issue (2): 220-226.DOI: 10.13304/j.nykjdb.2021.0728

• 方法与技术创新 • 上一篇

高效毛细管电泳法测定金针菇提取物中β-烟酰胺单核苷酸含量的初步研究

马晓颖(), 肖军, 陈珣, 龚娜, 刘国丽, 杨涛, 肇莹()

辽宁省农业科学院食用菌研究所，辽宁省食用菌优质栽培重点实验室，沈阳 110161

收稿日期:2021-08-23 接受日期:2022-01-07 出版日期:2023-02-15 发布日期:2023-05-17
通讯作者: 肇莹
作者简介:马晓颖 Email：mxy851001@126.com；
基金资助:
揭榜挂帅科技攻关专项(2021JH1/10400035);辽宁农业攻关及产业化项目(2020JH2/10200013);中央引导地方科技发展专项资金(2021010053-JH6/105);辽宁科技特派行动专项计划(2021JH5/10400035);沈阳市农村科技特派团专项(21-116-3-23);辽宁省农业科学院院长基金项目(2022QN2303)

Preliminary Study on Determination of β-nicotinamide Mononucleotide Content from Flammulina velutiper Extract by High Performance Capillary Electrophoresis

Xiaoying MA(), Jun XIAO, Xun CHEN, Na GONG, Guoli LIU, Tao YANG, Ying ZHAO()

Institute of Edible Mushrooms，Liaoning Academy of Agricultural Sciences； Key Laboratory of Quality Cultivation of Edible Mushrooms in Liaoning Province，Shenyang 110161，China

Received:2021-08-23 Accepted:2022-01-07 Online:2023-02-15 Published:2023-05-17
Contact: Ying ZHAO

摘要/Abstract

摘要：

金针菇既是一种美味食品，又是较好的保健食品，营养成分十分丰富，建立了金针菇提取液中β-烟酰胺单核苷酸（β?-nicotinamide mononucleotide，NMN）的高效毛细管电泳（high performance capillary electrophoresis，HPCE-DAD）含量测定方法。试验结果表明，检测波长218 nm，运行缓冲液30 mmol·L^-1硼砂缓冲液和10 mmol·L^-1磷酸二氢钠缓冲液（pH 9.2），分离电压25 kV，进样时间为5 s时，该方法能够检测到金针菇提取液中的β-烟酰胺单核苷酸，检测限为0.17 μg·mL^-1，并在0.03~2.00 mg·mL^-1范围内呈较好的线性关系，利用标准曲线获得回归方程y=2E+6x-345.69，R²=0.999 4，将金针菇图谱中的相应β-烟酰胺单核苷酸（NMN）峰面积带入回归方程，计算得β-烟酰胺单核苷酸含量为27.57 μg·mL^-1。方法学考察结果表明β-烟酰胺单核苷酸标准品迁移时间和峰面积相对偏差为0.68%和1.44%；且样品8 h内的迁移时间和峰面积的相对偏差分别为1.26%和2.82%；β-烟酰胺单核苷酸在金针菇样品中的回收率为89%~94%。结果表明该方法适用于金针菇提取液中β-烟酰胺单核苷酸的测定，为食用菌中β-烟酰胺单核苷酸含量的测定提供了新的方法。

关键词: 高效毛细管电泳, β-烟酰胺单核苷酸, 金针菇

Abstract:

Flammulina velutiper is not only a delicious food， but also a good health food which is rich in nutrition. However， there was no report on the determination of β?-nicotinamide mononucleotide （NMN） content in Flammulina velutiper. A method for the determination of β??-nicotinamide mononucleotide （NMN） in Flammulina velutiper was established by high performance capillary electrophoresis （HPCE-DAD）. The detection conditions included detection wavelength 218 nm； running buffer （30 mmol·L^-1 boron sand buffer pH 9.2； 10 mmol·L^-1 phosphate bobbin）； separation voltage of 25 kV； injection time of 5 s. The results showed that the method could detect the β-nicotinamide mononucleotide content of fruits from Flammulina velutiper with the detection limit of 0.17 μg·mL^-1， and showed a good linear relationship from 0.03 to 2.00 mg·mL^-1. The regression equation was obtained by the standard curve： y=2E+6x-345.69，R²=0.999 4， the corresponding β??-nicotinamide mononucleotide （NMN） peak area was put into the regression equation， and the calculated content was 27.57 μg·mL^-1. The relative standard deviations of migration times for β??-nicotinamide mononucleotide and migration peak areas were 0.68% and 1.44%. The relative deviations of migration time and peak area within 8 h were 1.26% and 2.82%， respectively. The recoveries of β??-nicotinamide mononucleotide in Flammulina velutiper were 89%~94%. These results indicated that the method was suitable for the determination of β??-nicotinamide mononucleotide in Flammulina velutiper and provided a new method for the determination of β-nicotinamide mononucleotide in edible fungus.

Key words: high performance capillary electrophoresis, β-nicotinamide mononucleotide, Flammulina velutiper

中图分类号:

S646

马晓颖, 肖军, 陈珣, 龚娜, 刘国丽, 杨涛, 肇莹. 高效毛细管电泳法测定金针菇提取物中β-烟酰胺单核苷酸含量的初步研究[J]. 中国农业科技导报, 2023, 25(2): 220-226.

Xiaoying MA, Jun XIAO, Xun CHEN, Na GONG, Guoli LIU, Tao YANG, Ying ZHAO. Preliminary Study on Determination of β-nicotinamide Mononucleotide Content from Flammulina velutiper Extract by High Performance Capillary Electrophoresis[J]. Journal of Agricultural Science and Technology, 2023, 25(2): 220-226.

图/表 5

图1 内标法测定β-烟酰胺单核苷酸的试验结果图谱

Fig. 1 Chromatogram of the determination of β-nicotinamide mononucleotide by internal standard method

图2 不同浓度的β-烟酰胺单核苷酸的电泳图谱

Fig. 2 Capillary electrophoresis of β-nicotinamide mononucleotide at different concentrations

图3 β-烟酰胺单核苷酸的标准曲线

Fig. 3 Standard curve of β-nicotinamide mononucleotide

表1 金针菇样品中添加β-烟酰胺单核苷酸的回收率

Table 1 Recovery of the samples added β-nicotinamide mononucleotide

标准品

Compound

原始含量Original content/（mg·mL^-1）

加标量Added content/（mg·mL^-1）

测得量Found content/（mg·mL^-1）

平均回收率

Recovery rate/%

β-烟酰胺单核苷酸

β-nicotinamide mononucleotide

27 573

9 191

34 589

β-烟酰胺单核苷酸

β-nicotinamide mononucleotide

27 573

50 236

β-烟酰胺单核苷酸

β-nicotinamide mononucleotide

27 573

82 719

98 192

图4 不同放置时间的β-烟酰胺单核苷酸毛细管电泳图谱

Fig. 4 Capillary electrophoresis of β-nicotinamide mononucleotide at different times

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高效毛细管电泳法测定金针菇提取物中β-烟酰胺单核苷酸含量的初步研究

Preliminary Study on Determination of β-nicotinamide Mononucleotide Content from Flammulina velutiper Extract by High Performance Capillary Electrophoresis

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