A porous graphitized carbon LC-ESI/MS method for the quantitation of metronidazole and fluconazole in breast milk and human plasma

J Chromatogr B Analyt Technol Biomed Life Sci. 2018 May 1:1084:175-184. doi: 10.1016/j.jchromb.2018.03.034. Epub 2018 Mar 22.


Information on drug transfer into the breast milk is essential to protect the infant from undesirable adverse effects of maternal consumption of drugs and to allow effective pharmacological treatment of breastfeeding mothers. Metronidazole and fluconazole are two drugs frequently used in nursing women to treat various infections, thus questioning infant's safety due to drug exposure through breast milk. In this article a porous graphitized carbon LC/ESI-MS assay was developed for the quantitation of metronidazole and fluconazole in breast milk and human plasma. The assay was based on the use of 150 μL of biological samples, following acetonitrile precipitation of proteins and filtration that enabled injection into the LC/ESI-MS system. All analytes and the internal standard, ropinirole, were separated by using a porous graphitized carbon analytical column (150 × 2.1 mm i.d., particle size 5 μm) with isocratic elution. The mobile phase consists of 55% acetonitrile in water acidified with 0.1% concentrated formic acid and pumped at a flow rate of 0.25 mL min-1. The assay was linear over a concentration range of 0.1 to 15 μg mL-1 for all analytes in both biological samples. Intermediate precision was found to be <8.4% over the tested concentration ranges. A run time of <5 min for each sample made it possible to analyze a large number of biological samples per day. The method is the first reported application for the analysis of metronidazole and fluconazole in both breast milk and human plasma and it can be used to support a wide range of clinical studies.

Keywords: Breast milk; Fluconazole; Human plasma; LC-ESI/MS; Metronidazole; Porous graphitized carbon.

MeSH terms

  • Acetonitriles
  • Chromatography, Liquid / methods*
  • Female
  • Fluconazole / analysis*
  • Graphite / chemistry
  • Humans
  • Limit of Detection
  • Linear Models
  • Male
  • Metronidazole / analysis*
  • Milk, Human / chemistry*
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization / methods*


  • Acetonitriles
  • Metronidazole
  • Graphite
  • Fluconazole
  • acetonitrile