title: Determination of methylglyoxal in human blood plasma using fluorescence high performance liquid chromatography after derivatization with 1,2-diamino-4,5-methylenedioxybenzene.
abstract: Methylglyoxal (MG) is a highly reactive dicarbonyl compound that promotes the non-enzymatic glycation of proteins to yield irreversible advanced glycated end products, leading to the cross-linking or degradation of proteins. The physiological relevance of MG currently remains unclear because its metabolic behavior has not yet been elucidated in detail. Although several labeling methods that require a HPLC system have been developed and used to measure MG, a standard method to analyze the content of MG in biological samples has not been established. We herein present a practical method based on HPLC with fluorescence detection to measure low MG levels. MG concentrations were also measured in human blood plasma using the present method in order to demonstrate its utility. A calibration curve was produced using freshly purified MG at concentrations ranging between 0.05 and 1.0μM. The intra-day and inter-day relative standard diviations of the method were 2.55% and 4.03%, respectively. The limit of detection and limit of quantification were 60fmol and 200fmol, respectively for MG with a 10-μl injection volume of the derivatized sample solution. When the optimized method was applied to human plasma, the resulting concentrations of MG in the plasma of healthy subjects (n=23) ranged between 0.024 and 0.258μM (mean±SD=0.098±0.066). Thus, the method developed herein is simple, sensitive, and easy to operate for the measurement of MG in biological samples.
authors.affiliation: Department of Analytical Biochemistry, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan. Electronic address: yo@my-pharm.ac.jp.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Ogasawara
authors.affiliation: Department of Analytical Biochemistry, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Tanaka
authors.affiliation: Department of Analytical Biochemistry, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan.
authors.forename: Shin
authors.initials: S
authors.lastname: Koike
authors.affiliation: Department of Psychiatry and Behavioral Sciences, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa, Setagaya, Tokyo, 156-8506, Japan.
authors.forename: Yasue
authors.initials: Y
authors.lastname: Horiuchi
authors.affiliation: Department of Psychiatry and Behavioral Sciences, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa, Setagaya, Tokyo, 156-8506, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Miyashita
authors.affiliation: Department of Psychiatry and Behavioral Sciences, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa, Setagaya, Tokyo, 156-8506, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Arai
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods*; D005260:Female; D005453:Fluorescence; D006801:Humans; D057230:Limit of Detection; D008297:Male; D008875:Middle Aged; D010655:Phenylenediamines / Q000737:chemistry; D011765:Pyruvaldehyde / Q000097:blood*; D015203:Reproducibility of Results
chemical_list: D010655:Phenylenediamines; C058886:1,2-diamino-4,5-methylenedioxybenzene; D011765:Pyruvaldehyde
keywords: Biomarker; Carbonyl stress; Fluorescence detection; HPLC method; Methylglyoxal