The adrenal gland is important for many physiological and pathophysiological processes, but studies are often restricted by limited availability of sample material. Improved methods for sample preparation are needed to facilitate analyses of multiple classes of adrenal metabolites and macromolecules in a single sample. A procedure was developed for preparation of chromaffin cells, mouse adrenals, and human chromaffin tumors that allows for multi-omics analyses of different metabolites and preservation of native proteins. To evaluate the new procedure, aliquots of samples were also prepared using conventional procedures. Metabolites were analyzed by liquid-chromatography with mass spectrometry or electrochemical detection. Metabolite contents of chromaffin cells and tissues analyzed with the new procedure were similar or even higher than with conventional methods. Catecholamine contents were comparable between both procedures. The TCA cycle metabolites, cis-aconitate, isocitate, and α-ketoglutarate were detected at higher concentrations in cells, while in tumor tissue only isocitrate and potentially fumarate were measured at higher contents. In contrast, in a broad untargeted metabolomics approach, a methanol-based preparation procedure of adrenals led to a 1.3-fold higher number of detected metabolites. The established procedure also allows for simultaneous investigation of adrenal hormones and related enzyme activities as well as proteins within a single sample. This novel multi-omics approach not only minimizes the amount of sample required and overcomes problems associated with tissue heterogeneity, but also provides a more complete picture of adrenal function and intra-adrenal interactions than previously possible.

中文翻译：

---

肾上腺激素相互作用和代谢：单一样本多组学方法

肾上腺对许多生理和病理生理过程都很重要，但研究往往受到样本材料有限可用性的限制。需要改进的样品制备方法来促进对单个样品中多类肾上腺代谢物和大分子的分析。开发了一种用于制备嗜铬细胞、小鼠肾上腺和人类嗜铬肿瘤的程序，允许对不同代谢物进行多组学分析并保存天然蛋白质。为了评估新程序，还使用常规程序制备了等分试样。代谢物通过液相色谱与质谱或电化学检测进行分析。使用新方法分析的嗜铬细胞和组织的代谢物含量与传统方法相似甚至更高。两种程序之间的儿茶酚胺含量相当。TCA 循环代谢物、顺乌头酸、异柠檬酸和 α-酮戊二酸在细胞中检测到的浓度较高，而在肿瘤组织中仅检测到异柠檬酸和潜在的富马酸含量较高。相比之下，在广泛的非靶向代谢组学方法中，基于甲醇的肾上腺制备程序导致检测到的代谢物数量增加了 1.3 倍。既定的程序还允许同时研究单个样品中的肾上腺激素和相关酶活性以及蛋白质。