Food and Fermentation Industries >

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

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

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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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

Cite this article

ZENG Xi , LUO Zhifeng , WANG Qiaoshan , SHEN Yudong , WANG Yu , XU Zhenlin . Rapid determination of melatonin in milk and formula with solid phase extraction-high performance liquid chromatography tandem mass spectrometry[J]. Food and Fermentation Industries, 2026 , 52(12) : 392 -398 . DOI: 10.13995/j.cnki.11-1802/ts.044641

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