Quantification of three beta-lactam antibiotics in breast milk and human plasma by hydrophilic interaction liquid chromatography/positive-ion electrospray ionization mass spectrometry

Drug Test Anal. 2017 Jul;9(7):1062-1072. doi: 10.1002/dta.2104. Epub 2016 Nov 4.


The use of cephalosporins during breast feeding raises several issues, including the risk of drug exposure through breast milk for the infant. In this paper, a hydrophilic interaction liquid chromatography/positive ion electrospray mass spectrometric assay (HILIC/ESI-MS) was developed for the quantitation of cefuroxime, cefoxitin, and cefazolin in breast milk and human plasma. The assay was based on the use of small sample size, 25 μL of biological samples, following acetonitrile precipitation of proteins and filtration that enabled injection into the HILIC/ESI-MS system. All analytes and the internal standard, alfuzosin, were separated by using a ZIC®-HILIC analytical column (150.0 × 2.1 mm i.d., particle size 3.5 µm, 200 Å) with isocratic elution. The mobile phase was composed of a 6% 12.5 mM ammonium acetate water solution in acetonitrile and pumped at a flow rate of 0.25 mL min-1 . The assay was linear over a concentration range of 0.2 to 5 µg mL-1 and 0.4 to 20 µg mL-1 for all the analytes in breast milk and in human plasma, respectively. Intermediate precision was found to be less than 4.2% over the tested concentration ranges. A run time of less than 12 min for each sample made it possible to analyze a large number of biological samples per day. The method is the first reported application of HILIC in the analysis of antibiotics in breast milk and human plasma and it can be used to support a wide range of clinical studies. Copyright © 2016 John Wiley & Sons, Ltd.

Keywords: antibiotics; breast milk; human plasma; hydrophilic interaction liquid chromatography; liquid chromatography/mass spectrometry.

Publication types

  • Validation Study

MeSH terms

  • Anti-Bacterial Agents / analysis*
  • Anti-Bacterial Agents / blood*
  • Cefazolin / analysis
  • Cefazolin / blood
  • Cefoxitin / analysis
  • Cefoxitin / blood
  • Cefuroxime / analysis
  • Cefuroxime / blood
  • Chromatography, Liquid / methods
  • Female
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Limit of Detection
  • Milk, Human / chemistry*
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • beta-Lactams / analysis*
  • beta-Lactams / blood*


  • Anti-Bacterial Agents
  • beta-Lactams
  • Cefoxitin
  • Cefazolin
  • Cefuroxime