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Steroid hormones are small molecules (MW around 300 Da) characterized by a large range of polarity and their analysis has always presented a serious challenge. Persistent problems with the specificity of conventional immunological methods are the cause of inconsistent results in the literature, a particularly problematic situation for healthcare decisions. At present, mass spectrometric methods have become the gold standard for accurate steroid profiling, and their advent will require the re-analysis of previously published data. However, it is a common misconception to consider the use of theses sophisticated technologies as a guarantee for accurate measures. Steroid analysis, especially in nervous tissues, indeed requires well-validated purification and separation steps before mass spectrometry, only then will mass spectrometric analysis be the absolute reference methodology.
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Lière et al. (2015) studied this question.
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