通过兔肝氧代醛脱氢酶将糖基化中间体3-脱氧葡萄糖酮氧化为2-酮-3-脱氧葡萄糖酸，然后进行高效液相色谱法对其进行定量分析。

Quantitation of the glycation intermediate 3-deoxyglucosone by oxidation with rabbit liver oxoaldehyde dehydrogenase to 2-keto-3-deoxygluconic acid followed by high-performance liquid chromatography.

作者信息

Fujii E, Iwase H, Ishii-Karakasa I, Yajima Y, Hotta K

机构信息

Department of Internal Medicine, School of Medicine, Kitasato University, Kanagawa, Japan.

出版信息

J Chromatogr B Biomed Appl. 1994 Oct 14;660(2):265-70. doi: 10.1016/0378-4347(94)00313-0.

DOI:10.1016/0378-4347(94)00313-0

PMID:7866516

Abstract

A simple and sensitive method for the detection of 3-deoxyglucosone was developed using oxidation with crude oxoaldehyde dehydrogenase to 2-keto-3-deoxygluconic acid followed by high-performance liquid chromatography (HPLC). Oxoaldehyde dehydrogenase was prepared from rabbit liver and partially characterized. 2-Keto-3-deoxygluconic acid produced from 3-deoxyglucosone by oxoaldehyde dehydrogenase was derivatized with 1,2-diamino-4,5-methylenedioxybenzene, and the fluorescent products were detected and quantitated by HPLC using a solvent containing borate. In the presence of borate, 2-keto-3-deoxygluconic acid was completely separated from N-acetylneuraminic acid. The detection limit of 3-deoxyglucosone was 2.5 pmol/injection (10 microliters) at a signal-to-noise ratio of 3. This method was used to confirm the inhibitory effect of aminoguanidine on glycation.

摘要

开发了一种简单灵敏的检测3-脱氧葡萄糖酮的方法，该方法先用粗制的氧化醛脱氢酶将3-脱氧葡萄糖酮氧化为2-酮基-3-脱氧葡萄糖酸，然后进行高效液相色谱（HPLC）分析。氧化醛脱氢酶从兔肝脏中制备并进行了部分特性鉴定。氧化醛脱氢酶作用于3-脱氧葡萄糖酮产生的2-酮基-3-脱氧葡萄糖酸用1,2-二氨基-4,5-亚甲二氧基苯进行衍生化，荧光产物通过使用含硼酸盐的溶剂的HPLC进行检测和定量。在硼酸盐存在下，2-酮基-3-脱氧葡萄糖酸与N-乙酰神经氨酸完全分离。3-脱氧葡萄糖酮的检测限在信噪比为3时为2.5 pmol/进样（10微升）。该方法用于证实氨基胍对糖基化的抑制作用。