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The adrenal gland is important for many physiological and pathophysiologicalprocesses, but studies are often restricted by limited availability of samplematerial. Improved methods for sample preparation are needed to facilitateanalyses of multiple classes of adrenal metabolites and macromolecules in asingle sample. A procedure was developed for preparation of chromaffin cells,mouse adrenals, and human chromaffin tumors that allows for multi-omics analysesof different metabolites and preservation of native proteins. To evaluate thenew procedure, aliquots of samples were also prepared using conventionalprocedures. Metabolites were analyzed by liquid-chromatography with massspectrometry or electrochemical detection. Metabolite contents of chromaffincells and tissues analyzed with the new procedure were similar or even higherthan with conventional methods. Catecholamine contents were comparable betweenboth procedures. The TCA cycle metabolites, cis-aconitate, isocitate, andα-ketoglutarate were detected at higher concentrations in cells, whilein tumor tissue only isocitrate and potentially fumarate were measured at highercontents. In contrast, in a broad untargeted metabolomics approach, amethanol-based preparation procedure of adrenals led to a 1.3-fold higher numberof detected metabolites. The established procedure also allows for simultaneousinvestigation of adrenal hormones and related enzyme activities as well asproteins within a single sample. This novel multi-omics approach not onlyminimizes the amount of sample required and overcomes problems associated withtissue heterogeneity, but also provides a more complete picture of adrenalfunction and intra-adrenal interactions than previously possible.

Adrenal Hormone Interactions and Metabolism: A Single Sample Multi-Omics Approach