Determination of\nFertility Hormones and Adipokines\nby LC-MRM/MS Analysis

Due to the increase\nin the rate of male and female infertility,\nassisted fertilization practices are currently adopted as valid support\nfor couples unable to get pregnant. Analytical approaches for fertility\nhormone dosages are constantly being developed, following the technological\nprogress of fertilization methods that have evolved for more than\na century. Indeed, the analysis of fertility hormones in serum samples\nis a common clinical practice to check the fertility state, but absolute\nquantification of these hormones is a great challenge due to biological\nvariability and low serum concentrations. Currently, ELISA (enzyme-linked\nimmunosorbent assay) based methods are the most used analytical techniques\nto quantify hormones in blood in clinical settings. The current Article\ndiscusses the development of a liquid chromatography-tandem mass spectrometry\nmethod (LC-MS/MS) to monitor multiple fertility hormones of a protein\nnature in a single chromatographic run, i.e., LH (luteinizing hormone),\nFSH (follicle-stimulating hormone), TSH (thyroid-stimulating hormone),\nAMH (anti-Müllerian hormone), adiponectin, ghrelin, leptin,\nglucagon, and obestatin. Particular attention has been paid to the\nAMH hormone, whose ELISA-based quantification is known to be controversial\ndue to the poor reproducibility between the various kits used. For\nAMH, the internal standard method was used for the quantitative determination\nto compare mass spectrometry data to the ELISA assays performed by\nan accredited analysis laboratory on a cohort of samples from women\naged between 18 and 60 years. The ability to monitor multiple transitions\nby LC-MRM/MS ensured both high specificity and high selectivity,\nwhich is necessary for the quantification of protein and steroid hormones,\nbesides improvements in data reproducibility and reduced analysis\ntimes and costs.