Biosci. Biotechnol. Biochem., 71 (9), 2162–2168, 2007
Determination of Glyceraldehyde Formed in Glucose Degradation
and Glycation
y
1;
Teruyuki USUI,1;2 Miku YOSHINO,1 Hirohito WATANABE,3 and Fumitaka HAYASE
1Department of Agricultural Chemistry, Faculty of Agriculture, Meiji University,
1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan
2Department of Health Care Nutrition, Showagakuin Junior College,
2-17-1 Higashisugano, Ichikawa, Chiba 272-0823, Japan
3Department of Life Sciences, Faculty of Agriculture, Meiji University,
1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan
Received February 7, 2007; Accepted May 23, 2007; Online Publication, September 7, 2007
Glyceraldehyde (GLA) was determined in glucose de-
gradation and glycation. GLA was detected as a deca-
hydroacridine-1,8-dione derivative on reversed phase
HPLC using cyclohexane-1,3-dione derivatizing re-
agent. The glucose-derived GLA level was higher than
the glycation-derived GLA level, because GLA was con-
verted to intermediates and advanced glycation end
products (AGE) in glycation. GLA was also generated
from 3-deoxyglucosone and glucosone as intermediates
of glucose degradation and glycation. This study sug-
gests that glyceraldehyde is generated by hyperglycemia
in diabetes, and that it is also formed in medicines such
as peritoneal dialysis solution.
GLA and GLA-derived AGE are generated in some
diseases, and can cause the pathological changes and
physiological effects. GLA is perhaps increased by
reduced activity of GLA-related enzymes, such as
glyceraldehyde-3-phosphate dehydrogenase (GAPDH).
In addition, GLA might be generated by glucose
degradation and the glycation.
In this study, GLA, generated in glucose degradation
and glycation, was determined to be a decahydroacri-
dine-1,8-dione derivative by reversed phase HPLC by
simple derivatization using cyclohexane-1,3-dione.
Materials and Methods
Key words: glyceraldehyde; 1,3-cyclohexanedione; gly-
cation; Maillard reaction; glucose degrada-
tion
Chemicals. Glyceraldehyde and glucose were ob-
tained from Sigma-Aldrich (St. Louis, MO). Cyclo-
hexane-1,3-dione and benzoylhydrazine were from
Tokyo Chemical Industry (Tokyo). Benzaldehyde, ace-
tic acid, and methanol (HPLC grade) were from Kanto
Chemical (Tokyo). Glycine, lysine, p-toluidine, and 2,4-
dinitrophenylhydrazine were from Wako Pure Chemical
(Osaka, Japan). All other chemicals used in this study
were of analytical reagent grade. Water was used after
purification by reverse osmosis, using Milli RO 10PLUS
(Millipore, Billerica, MA), and further treated with
Simpli Lab-UV (Millipore).
Carbonyl compounds cause glycation (the Maillard
reaction) in post-translational modification processes.1,2)
Recent studies suggest that glyceraldehyde-modified
protein affects cell function. GLA-modified albumin
shows cytotoxicity in rat primary cultured cortical neu-
rons3) and Schwann cells.4) In addition, GLA-modified
albumin induces apoptosis and expression of vascular
endothelial growth factor (VEGF) in bovine retinal
pericytes,5) and also induces apoptosis and expression of
VEGF and monocyte chemoattractant protein-1 (MCP-
1) in human mesangial cells.6) We have reported that
GLA-derived advanced glycation end product (AGE)
induces oxidative stress.7,8) Moreover, GLA-modified
protein is detected by an immunological technique in the
Alzheimer’s disease specimens.9)
Synthesis of 3-deoxyglucosone. 3-Deoxyglucosone
(3DG) was synthesized according to the method de-
scribed by Kato et al.10) D-glucose (16 g) and p-toluidine
(8.8 g) were refluxed in a mixture of 95% ethanol
(360 ml) and acetic acid (17.6 ml) at 110 ꢀC for 30 min.
Then benzoylhydrazine (26.4 g) was added to the sol-
y
To whom correspondence should be addressed. Fax: +81-44-934-7902; E-mail: fumi@isc.meiji.ac.jp
Abbreviations: AGE, advanced glycation end products; ANOVA, one factor analysis of variance; CAPD, continuous ambulatory peritoneal
dialysis; 1,3CHD, cyclohexane-1,3-dione; DA, dihydroxyacetone; 3DG, 3-deoxyglucosone; DNP, 2,4-dinitrophenylhydrazine; GLA, glyceraldehyde;
GLUCO, glucosone; MCP-1, monocyte chemoattractant protein-1; MGO, methylglyoxal; PDA, photo diode array; TR, triose reductone; TRIO,
triosone; VEGF, vascular endothelial growth factor