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)

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• 作者：Romain Simon ; Guillaume Jubeaux ; Arnaud Chaumot ; Jé ; rô ; me Lemoine ; Olivier Geffard ; Arnaud Salvador
• 关键词：Biomarkers ; Protein quantification ; Crustacean ; Mass spectrometry ; Vitellogenin ; Endocrine disruptor ; MRM
• 刊名：Journal of Chromatography A
• 出版年：2010
• 出版时间：30 July 2010
• 年：2010
• 卷：1217
• 期：31
• 页码：5109-5115
• 全文大小：311 K

文摘

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 2.5–25,000 pg/mL and the limit of detection was estimated at 0.75 pg/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.