Study of transaldolase deficiency in urine samples by capillary LC-MS/MS.
Transaldolase (TAL) is a key enzyme of the pentose phosphate pathway (PPP). TAL deficiency is a newly recognized cause of liver cirrhosis. We have developed an ion-pair LC separation combined with negative ion electrospray MS/MS detection method to assess PPP metabolites in urine samples from TAL-deficient mice. Sedoheptulose 7-phosphate (S7P), C5-polyols D-arabitol and D-ribitol, and 6-phosphogluconate (6PG) levels were markedly increased in urine of TAL-deficient mice with respect to those of wild-type and heterozygote littermates. The detection limits of S7P, D-arabitol, and 6PG were 0.15 +/- 0.015 pmol, 3.5 +/- 0.41 pmol, and 0.61 +/- 0.055 pmol, respectively. The limit of quantitation was 0.4 +/- 0.024 nmol/ml for S7P, 1.6 +/- 0.11 nmol/ml for 6PG and 10 +/- 0.7 nmol/ml for D-arabitol. Additional metabolites, hexose 6-phosphates (m/z 259), D-ribose 5-phosphate and D-xylulose 5-phosphate (m/z 229), D-fructose 1,6-diphosphate (m/z 339), C6-polyols (m/z 181) and GSSG (m/z 611), that have been positively identified in mouse urine, showed similar levels in control and TAL-deficient mice.
SUNY Upstate Medical University, Department of Medicine, NY 13210, USA.
György
G
Vas
Karina
K
Conkrite
Wendy
W
Amidon
Yueming
Y
Qian
Katalin
K
Bánki
András
A
Perl
D000818:Animals; D053801:Capillary Electrochromatography / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D010427:Pentose Phosphate Pathway; D013403:Sugar Phosphates / Q000652:urine*; D014153:Transaldolase / Q000737:chemistry* / Q000378:metabolism*; D016482:Urinalysis / Q000379:methods*
D013403:Sugar Phosphates; D014153:Transaldolase