Analytical and Bioanalytical Chemistry

, Volume 405, Issue 26, pp 8645–8652 | Cite as

Rapid analysis of aflatoxin M1 in milk using dispersive liquid–liquid microextraction coupled with ultrahigh pressure liquid chromatography tandem mass spectrometry

  • Luca Campone
  • Anna Lisa PiccinelliEmail author
  • Rita Celano
  • Mariateresa Russo
  • Luca Rastrelli
Research Paper


A simple, rapid, and sensitive method based on simultaneous protein precipitation and extraction of aflatoxin M1 (AFM1) followed by dispersive liquid–liquid microextraction (DLLME) and ultrahigh pressure liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) analysis was developed for the determination of AFM1 in milk samples. In order to precipitate the proteins and extract AFM1 from milk, a sample pretreatment using acetonitrile and NaCl as the extraction/denaturant solvent and salting-out agent, respectively, was optimised. Subsequently, the acetonitrile (upper) phase, containing AFM1, was used as the disperser solvent in DLLME, and extractant (chloroform) and water were added in turn to the extract to perform the DLLME process. The main parameters affecting the extraction efficiency of the whole analytical procedure, such as acetonitrile volume, amount of salt, type and volume of extractant and water volume, were carefully optimised by experimental design. Under optimum conditions, the developed method provides an enrichment factor of 33 and detection and quantification limits (0.6 and 2.0 ng kg−1, respectively) below the maximum levels imposed by current regulations for AFM1 in milk and infant milk formulae. Recoveries (61.3–75.3 %) and repeatability (RSD < 10, n = 3), tested in different types of milk at four AFM1 levels, met the performance criteria required by EC Regulation No. 401/2006. Moreover, the matrix effect on the signal intensity of the analyte was negligible. The proposed method provides a rapid extraction and an accurate determination of AFM1 in milk and formula milk using a simple and inexpensive sample preparation procedure.


Rapid analysis of aflatoxin M1 in milk using DLLME


Milk Aflatoxin M1 Protein precipitation Dispersive liquid–liquid microextraction UHPLC-MS/MS 



The Project was funded by the Italian Ministry of the University and Research (MIUR) with a FIRB “Futuro in Ricerca” Project n. RBFR10GSJK “Tecniche Analitiche Avanzate per l’Analisi dei Contaminanti negli Alimenti”.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Luca Campone
    • 1
  • Anna Lisa Piccinelli
    • 1
    Email author
  • Rita Celano
    • 1
  • Mariateresa Russo
    • 2
  • Luca Rastrelli
    • 1
  1. 1.Dipartimento di FarmaciaUniversità di SalernoFisciano (SA)Italy
  2. 2.Dipartimento di AgrariaUniversità Mediterranea di Reggio CalabriaReggio di CalabriaItaly

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