Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Piazzale Aldo Moro, 00185, Rome, Italy.
Department of Cell Biology and Neuroscience, Istituto Superiore di Sanità, Viale Regina Elena, Rome, Italy.
Adv Exp Med Biol. 2017;975 Pt 1:535-549. doi: 10.1007/978-94-024-1079-2_42.
There is an increasing interest for analytical methods aimed to detect biological sulfur-containing amines, because of their involvement in human diseases and metabolic disorders. This work describes an improved HPLC method for the determination of sulfur containing amino acids and amines from different biological matrices. We optimized a pre-column derivatization procedure using dabsyl chloride, in which dabsylated products can be monitored spectrophotometrically at 460 nm. This method allows the simultaneous analysis of biogenic amines, amino acids and sulfo-amino compounds including carnosine, dopamine, epinephrine, glutathione, cysteine, taurine, lanthionine, and cystathionine in brain specimens, urines, plasma, and cell lysates. Moreover, the method is suitable for the study of physiological and non-physiological derivatives of taurine and glutathione such as hypotaurine, homotaurine, homocysteic acid and S-acetylglutathione. The present method displays good efficiency of derivatization, having the advantage to give rise to stable products compared to other derivatizing agents such as o-phthalaldehyde and dansyl chloride.With this method, we provide a tool to study sulfur cycle from a metabolic point of view in relation to the pattern of biological amino-compounds, allowing researchers to get a complete scenario of organic sulfur and amino metabolism in tissues and cells.
由于含硫生物胺在人类疾病和代谢紊乱中的作用，人们对其分析方法的兴趣日益增加。本工作描述了一种改进的 HPLC 方法，用于测定来自不同生物基质的含硫氨基酸和胺。我们优化了使用二甲氨基萘磺酰氯（dabsyl chloride）的柱前衍生程序，其中衍生产物可以在 460nm 处进行分光光度法监测。该方法可同时分析生物胺、氨基酸和磺基氨基酸化合物，包括脑标本、尿液、血浆和细胞裂解物中的肌肽、多巴胺、肾上腺素、谷胱甘肽、半胱氨酸、牛磺酸、硫代高丝氨酸和胱硫醚。此外，该方法适用于研究牛磺酸和谷胱甘肽的生理和非生理衍生物，如次牛磺酸、同型牛磺酸、高半胱氨酸和 S-乙酰谷胱甘肽。与其他衍生试剂如邻苯二醛和丹磺酰氯相比，该方法具有良好的衍生效率优势，可产生稳定的产物。通过这种方法，我们从代谢角度提供了一种研究硫循环的工具，与生物氨基酸化合物的模式相关联，使研究人员能够在组织和细胞中全面了解有机硫和氨基酸代谢。
