S-Adenosylmethionine (SAM) and S-Adenosylhomocysteine (SAH) Monitoring Using Analytical Methods in Clinical Laboratory Practice: Where Are We?

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Abstract
S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) are essential intermediates in one-carbon metabolism and key regulators of cellular methylation capacity. Their concentrations and the SAM/SAH ratio are increasingly studied as biomarkers across metabolic, cardiovascular, neurological, and cancer-related diseases. This review outlines validated analytical methods for quantifying SAM and SAH, focusing primarily on liquid chromatography–tandem mass spectrometry (LC–MS/MS), which is considered the gold standard in both clinical and research settings. A comprehensive literature search identified studies on method development, validation, and clinical use of SAM and SAH measurements. Special attention is given to analytical challenges arising from their high polarity, structural similarity, endogenous presence, and limited stability. The review also discusses preanalytical variables, including biological matrix selection, sample handling, and storage conditions. LC–MS/MS methods are compared with alternative techniques, such as immunoassays, with respect to sensitivity, specificity, matrix effects, and clinical relevance. Additionally, the review summarizes the concentration ranges of SAM and SAH, and their ratio, in healthy and patient populations, noting current standardization limitations. Overall, the review highlights the importance of harmonized analytical protocols and matrix-specific validation to enable reliable clinical interpretation of SAM and SAH as methylation biomarkers.

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