超高效液相色谱-串联质谱法测定蜂王幼虫中烟酰胺单核苷酸的含量  

Determination of nicotinamide mononucleotide in queen bee larva by ultra-high performance liquid chromatographic tandem mass spectrometry

作　　者：赵彤 赵洪木[2] 宋美洁 杜伟 吴黎明[1] 范芳芳 薛晓锋[1] ZHAO Tong;ZHAO Hongmu;SONG Meijie;DU Wei;WU Liming;FAN Fangfang;XUE Xiaofeng(Institute of Apicultural Research,Chinese Academy of Agricultural Sciences,Beijing 100093,China;Sericultural and Apicultural Research Institute Yunnan Academy of Agricultural Sciences,Hani-Yi Autonomous Prefecture of Honghe 661100,China;Agilent Technologies(China)Co.Ltd.,Beijing 100102,China;Lao Dian Huang Institute of Life Sciences,Suzhou 215000,China)

机构地区：[1]中国农业科学院蜜蜂研究所,北京100093 [2]云南省农业科学院桑蚕蜜蜂研究所,云南红河哈尼族彝族自治州661100 [3]安捷伦科技(中国)有限公司,北京100102 [4]老滇凰生命科学院,江苏苏州215000

出　　处：《食品与发酵工业》2022年第8期265-269,共5页Food and Fermentation Industries

基　　金：所企合作创新项目(MFSHTK-CS-2020088);所企合作创新项目(202005AF150027)。

摘　　要：建立了超高效液相色谱-串联质谱快速测定蜂王幼虫中烟酰胺单核苷酸(nicotinamide mononucleotide,NMN)的方法。样品经乙醇和水提取后,采用Waters X Select HSS T_(3)(2.1 mm×100 mm,2.5μm)色谱柱进行分离,串联质谱多反应监测模式定量分析。该方法中NMN在50~1000 ng/mL线性关系良好(R^(2)=0.9997),检出限和定量限分别为11.2μg/kg和37.4μg/kg。在添加水平为20、40、80μg/g时,样品的回收率为109.9%~111.2%,相对标准偏差为0.87%~3.31%。应用方法测定来自不同蜂场的蜂王幼虫样品中NMN的含量,11个新鲜幼虫样品均检出NMN,其含量为37.0~65.8μg/g,1个蜂王幼虫冻干粉中NMN的含量为78.0μg/g。稳定性实验结果显示,蜂王幼虫中NMN极易降解,在常温和4℃条件贮存7 d的降解率分别为96.5%和88.5%,低温或加工成冻干粉可有效抑制NMN的降解。该方法适用于蜂王幼虫中NMN的检测,实际样品分析结果显示,NMN可作为蜂王幼虫及其产品品质评价的潜在指标。A simple and fast method for the determination of nicotinamide mononucleotide(NMN) in queen bee larva by ultra-high performance liquid chromatographic tandem mass spectrometry(UHPLC-MS/MS) was developed in this study. The NMN was separated on Waters X Select HSS T_(3)column(2.1 mm×100 mm, 2.5 μm) and quantified on multiple reaction monitor(MRM). The method was validated using spiked samples with NMN standard of 20, 40 and 80 μg/g. Recoveries were 109.9%-111.2% with relative standard deviation(RSD) 0.87%-3.31%. The calibration curve was fitted with the correlation coefficient R^(2)= 0.999 7 in the range of 50-1 000 ng/mL. The limits of detection(LOD) and the limits of quantitation(LOQ) were 11.2 and 37.4 μg/kg, respectively. The proposed UHPLC-MS/MS method was applied to the analysis of queen bee larva samples. The results showed that the NMN in 11 fresh queen bee larva samples was from 37.0 to 65.8 μg/g, especially in 1 freeze-dried powder sample which was 78.0 μg/g. The NMN in queen bee larva was easily broken down, and the degradation rates at room temperature and 4 ℃ for 7 d were 96.5% and 88.5%, respectively. Low temperature storage or lyophilization could prevent the degradation of NMN in queen bee larva. As a functional and nutritional component, NMN can be used as a potential indicator for the quality evaluation of queen bee larva and their products.

关 键 词：蜂王幼虫 烟酰胺单核苷酸 超高效液相色谱-串联质谱 

分 类 号：S896.7[农业科学—特种经济动物饲养] O657.63[农业科学—畜牧兽医]