Analytical and Bioanalytical Chemistry

, Volume 382, Issue 4, pp 1084–1092 | Cite as

Determination of phthalate monoesters in human milk, consumer milk, and infant formula by tandem mass spectrometry (LC–MS–MS)

  • Gerda K. Mortensen
  • Katharina M. Main
  • Anna-Maria Andersson
  • Henrik Leffers
  • Niels E. Skakkebæk
Original Paper


Daily exposure of humans to phthalates may be a health risk because animal experiments have shown these compounds can affect the differentiation and function of the reproductive system. Because milk is the main source of nutrition for infants, knowledge of phthalate levels is important for exposure and risk assessment. Here we describe the development and validation of a quantitative analytical procedure for determination of phthalate metabolites in human milk. The phthalate monoesters investigated were: monomethyl phthalate (mMP), monoethyl phthalate (mEP), mono-n-butyl phthalate (mBP), monobenzyl phthalate (mBzP), mono-(2-ethylhexyl) phthalate (mEHP), and monoisononyl phthalate (mNP). The method is based on liquid extraction with a mixture of ethyl acetate and cyclohexane (95:5) followed by two-step solid-phase extraction (SPE). Detection and quantification of the phthalate monoesters were accomplished by high-pressure liquid chromatography using a Betasil phenyl column (100 mm×2.1 mm×3 μm) and triple tandem mass spectrometry (LC–MS–MS). Detection limits were in the range 0.01 to 0.5 μg L−1 and method variation was from 5 to 15%. Analysis of 36 milk samples showed that all these phthalates were present, albeit at different concentrations. Median values (μg L−1) obtained were 0.11 (mMP), 0.95 (mEP), 3.5 (mBP), 0.8 (mBzP), 9.5 (mEHP), and 101 (mNP). We also analysed seven samples of consumer milk and ten samples of infant formula. Only mBP and mEHP were detected in these samples, in the ranges 0.6–3.9 μg L−1 (mBP) and 5.6–9.9 μg L−1 (mEHP).


Phthalates Exposure Analysis Human milk Consumer milk Infant formula 



The authors gratefully acknowledge financial support from Velux Fonden for instrumental equipment and financial support from the European Union, project Expored (QLK4-CT-2001-00269), and The Danish Medical Research Council (9700833, 9700909). We also thank John W. Brock PhD for his advice during the course of this project.


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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Gerda K. Mortensen
    • 1
  • Katharina M. Main
    • 1
  • Anna-Maria Andersson
    • 1
  • Henrik Leffers
    • 1
  • Niels E. Skakkebæk
    • 1
  1. 1.Department of Growth and ReproductionCopenhagen University Hospital CopenhagenDenmark

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