Mass spectrometry assay as an alternative to the enzyme-linked immunosorbent assay test for biomarker quantitation in ecotoxicology: application to vitellogenin in Crustacea (Gammarus fossarum).
Vitellogenin (Vg) is a widespread biomarker for measuring exposure to endocrine disruptors. Vg quantification is usually done by using the ELISA test (enzyme-linked immunosorbent assay). Since this test is specific to a target protein, it can rarely be used with other species due to low cross-reactivity across species. Therefore alternative analytical methods have to be considered as the development of a specific and sensitive ELISA test for each new target is time-consuming and may prove unsuccessful. This paper focuses on the development of a quantitative assay by liquid chromatography tandem mass spectrometry (LC-MS/MS) of vitelogenin in an invertebrate (Gammarus fossarum) for which no ELISA kit is available. The linearity of the method was within the concentration range 2.5-25,000pg/mL and the limit of detection was estimated at 0.75pg/mL of Vg. This method has been demonstrated to be an alternative to existing immunological methods for quantifying Vg in invertebrates due to its greater sensitivity, specificity and reproducibility (intra- and inter-assay below 15%). This assay was applied for Vg determination in female G. fossarum following exposure to a known endocrine disruptor, methyl farnesoate, in crustaceans. The availability of a quantitative G. fossarum LC-MS/MS assay should open the way for further studies to evaluate xenoestrogen effects in aquatic male G. fossarum.
Simon
Romain
R
UMR 5180 Sciences Analytiques, Bât. CPE, Université de Lyon, Lyon 1, 69622 Villeurbanne cedex, France.
Jubeaux
Guillaume
G
Chaumot
Arnaud
A
Lemoine
Jérôme
J
Geffard
Olivier
O
Salvador
Arnaud
A
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002853:Chromatography, Liquid / Q000379:methods*; D003445:Crustacea / Q000737:chemistry* / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005231:Fatty Acids, Unsaturated / Q000633:toxicity*; D005260:Female; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D014819:Vitellogenins / Q000032:analysis*
D015415:Biomarkers; D005231:Fatty Acids, Unsaturated; D014819:Vitellogenins; C032429:methyl farnesoate