Systems Biology and Medicine Center for Complex Diseases, Center for Clinical Research, The Affiliated Hospital of Qingdao University, Qingdao, China.
Methods Mol Biol. 2022;2303:111-119. doi: 10.1007/978-1-0716-1398-6_10.
Glycosaminoglycan (GAG) fine structures from the same animal cells and tissues are controlled not only by the biosynthetic and metabolic enzymes but also by other environmental factors, such as chemicals, growth factors, nutritional factors, and isolation procedures. To facilitate direct quantitative comparison of disaccharide compositions from different GAG preparations, several stable isotope labeling strategies have been developed. In this report, 1-phenyl-3-methyl-5-pyrazolone (PMP) and deuterated d5-PMP are used for differential disaccharide labeling and profiling of chondroitin sulfate GAG by high performance liquid chromatography (HPLC) coupled with mass spectrometry (MS).
糖胺聚糖 (GAG) 的精细结构不仅受到生物合成和代谢酶的控制,还受到其他环境因素的影响,如化学物质、生长因子、营养因素和分离程序。为了便于直接定量比较不同 GAG 制剂的二糖组成,已经开发了几种稳定同位素标记策略。在本报告中,使用 1-苯基-3-甲基-5-吡唑酮 (PMP) 和氘代 d5-PMP 进行差异二糖标记,并通过高效液相色谱 (HPLC) 与质谱 (MS) 联用对硫酸软骨素 GAG 进行分析。