A sensitive enzymatic assay for the determination of sucrose in serum and urine.
BACKGROUND
Sucrose permeability has been suggested as a simple and non-invasive marker of gastric mucosal damage. We here report on a sensitive enzymatic assay using four sequential enzyme reactions coupled with reduced thio-NADPH.


METHODS
Sucrose is phosphorylated by sucrose phosphorylase (EC2.4.1.7). The subsequent reaction in the presence of phosphoglucomutase (EC5.4.2.2) and glucose-1,6-diphosphate forms glucose-6-phosphate. Sucrose of the monad forms the dyad thio-NADPH. The reaction is monitored by changes in absorbance at 405 nm.


RESULTS
The lower limit of detection (3SD method) was 2.8 micromol/l for serum and 7.0 micromol/l for urine. The precision of the method was <4.0%, and has sufficient analytical range.


CONCLUSIONS
The assay was sensitive enough to monitor serum sucrose concentrations during the sucrose permeability test and an automated assay may be useful in a large number of subjects.
Seimiya
Masanori
M
Division of Laboratory Medicine, Chiba University Hospital, 1-8-1 Inohana, Chuo, Chiba City, Chiba 260-8677, Japan. seichan@ho.chiba-u.ac.jp
Osawa
Susumu
S
Hisae
Nobuo
N
Shishido
Tadayuki
T
Yamaguchi
Taketo
T
Nomura
Fumio
F
D005753:Gastric Mucosa / Q000378:metabolism / Q000473:pathology; D005964:Glucosyltransferases / Q000378:metabolism; D006801:Humans; D007410:Intestinal Diseases / Q000097:blood / Q000378:metabolism / Q000473:pathology / Q000652:urine; D009249:NADP / Q000031:analogs & derivatives / Q000378:metabolism; D010539:Permeability; D010733:Phosphoglucomutase / Q000378:metabolism; D012680:Sensitivity and Specificity; D013395:Sucrose / Q000097:blood* / Q000378:metabolism / Q000652:urine*
D009249:NADP; D013395:Sucrose; D005964:Glucosyltransferases; C025901:sucrose phosphorylase; D010733:Phosphoglucomutase