食品与发酵工业 >

2026 , Vol. 52 >Issue 12: 392 - 398

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.044641

分析与检测

固相萃取-高效液相色谱-串联质谱法快速测定牛乳及乳粉中的褪黑素

  • 曾羲 ,
  • 罗志锋 ,
  • 王乔山 ,
  • 沈玉栋 ,
  • 王宇 ,
  • 徐振林
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  • 1(广州市食品检验所,广东 广州,511400)
    2(华南农业大学 食品学院,广东省食品质量安全重点实验室,广东 广州,510642)
    3(湖南农业大学 食品科学技术学院,湖南 长沙,410128)
第一作者:博士研究生,高级工程师(徐振林教授为通信作者,E-mail:jallent@163.com)

收稿日期: 2025-10-14

  修回日期: 2025-11-27

  网络出版日期: 2026-07-17

基金资助

广东省基础与应用基础研究基金项目(2023A1515012605)

收起

Rapid determination of melatonin in milk and formula with solid phase extraction-high performance liquid chromatography tandem mass spectrometry

  • ZENG Xi ,
  • LUO Zhifeng ,
  • WANG Qiaoshan ,
  • SHEN Yudong ,
  • WANG Yu ,
  • XU Zhenlin
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  • 1(Guangzhou Institute for Food Inspection, Guangzhou 511400, China)
    2(College of Food Science, South China Agricultural University, Guangdong Provincial Key Laboratory of Food Quality and Safety, Guangzhou 510642, China)
    3(College of Food Science and Technology, Hunan Agricultural University, Changsha 410128, China)

Received date: 2025-10-14

  Revised date: 2025-11-27

  Online published: 2026-07-17

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摘要

褪黑素因其具有调节昼夜规律、改善睡眠等功效,被用于功能性乳制品开发。该研究建立了1种基于固相萃取-高效液相色谱-串联质谱(solid phase extraction-high performance liquid chromatography tandem mass spectrometry,SPE-HPLC-MS/MS)技术快速测定牛乳和乳粉中褪黑素的分析方法。样品经蛋白沉淀、超声提取、离心和Oasis HLB柱净化、洗脱,采用电喷雾正离子(positive electrospray ionization,ESI+)、多反应监测(multiple reaction monitoring, MRM)模式,以水-乙腈为流动相,经C18柱(2.1 mm×100 mm×1.7 μm)柱梯度洗脱,外标法定量。结果表明,褪黑素在0.01~10.0 ng/mL线性关系良好,2条标准曲线的相关系数(R)均在0.995以上,样品检出限为0.005 ng/mL、定量限为0.020 ng/mL,在0.2、0.4、2.0 ng/g的3个加标水平上,乳粉和牛乳的日内加标回收率分别为93.1%~101.5%和94.4%~103.7%,日间加标回收率分别为90.5%～102.8%和92.3%～104.6%,相对标准偏差(relative standard deviations, RSDs)为2.5%~6.9%(n=6)。该方法操作简便、高效、灵敏度高,可用于牛乳和乳粉中微量褪黑素的测定,为功能性乳制品开发和监管提供技术支持。

关键词： 褪黑素; 乙酰基-5-甲氧基色胺; 固相萃取; 液相色谱-串联质谱; 乳粉; 牛乳

本文引用格式

曾羲 , 罗志锋 , 王乔山 , 沈玉栋 , 王宇 , 徐振林 . 固相萃取-高效液相色谱-串联质谱法快速测定牛乳及乳粉中的褪黑素[J]. 食品与发酵工业, 2026 , 52(12) : 392 -398 . DOI: 10.13995/j.cnki.11-1802/ts.044641

Abstract

Melatonin has been applied in the development of functional dairy products, attributed to its efficacy in regulating the circadian rhythm and improving sleep quality.This study set up a method based on solid phase extraction-high performance liquid chromatography tandem mass spectrometry in rapid determination of melatonin in milk and formula.Samples were processed by protein precipitation, then extracted by ultrasound and centrifugation.The supernatant was purified and eluted by Oasis HLB column.The eluent was conducted in positive electrospray ionization (ESI+) mode under multiple reaction monitoring (MRM) mode, coupled with gradient elution by water and acetonitrile as mobile phase and C18 chromatographic column (2.1 mm×100 mm×1.7 μm) chromatographic column, quantified by external standard.As a result, the linearity were good in the range of 0.01-10.0 ng/mL, with the regression coefficients (R) in two standard curves were both above 0.995.The limit of detection (LOD) was 0.005 ng/mL and the limit of quantitation (LOQ) was 0.020 ng/mL in real samples.The within-day recoveries of three spiked levels of milk powder and milk were 93.1%-101.5% and 94.4%-103.7%, and between-day recoveries were 90.5%-102.8% and 92.3%-104.6%, and the relative standard deviations were 2.5%-6.9% (n=6).This method is easy, effective and high-sensitive, which can be applied in melatonin determination for milk powder and milk.It can supply important real meaning for the development of functional milk products and technical support of milk market in China.

Key words： melatonin; N-acetyl-5-methoxytryptamine; solid phase extraction; liquid chromatography tandem mass spectrometry; milk powder; milk

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