Profiling of pentose phosphate pathway intermediates in blood spots by tandem mass spectrometry: application to transaldolase deficiency.
BACKGROUND
Recently, several patients with abnormal polyol profiles in body fluids have been reported, but the origins of these polyols are unknown. We hypothesized that they are derived from sugar phosphate intermediates of the pentose phosphate pathway (PPP), and we developed a semiquantitative method for profiling of pentose phosphate pathway intermediates.


METHODS
Sugar phosphates in blood spots were simultaneously analyzed by liquid chromatography-tandem mass spectrometry using an ion-pair-loaded C(18) HPLC column. The tandem mass spectrometer was operated in the multiple-reaction monitoring mode. Enzymatically prepared D-[(13)C(6)]glucose 6-phosphate was used as internal standard. The method was used to study sugar phosphates abnormalities in a patient affected with a deficiency of transaldolase (TALDO1; EC 2.2.1.2).


RESULTS
In control blood spots, dihydroxyacetone phosphate, pentulose 5-phosphates, pentose 5-phosphates, hexose 6-phosphates, and sedoheptulose 7-phosphate were detected. Detection limits ranged from approximately 100 to approximately 500 nmol/L. Glyceraldehyde 3-phosphate and erythrose 4-phosphate were undetectable. Intra- and interassay imprecision (CVs) were 10-17% and 12-21%, respectively. In blood from the TALDO1-deficient patient, sedoheptulose 7-phosphate was increased.


CONCLUSIONS
The new method allows investigation of patients in whom a defect in the PPP is suspected. Measurements of sugar phosphate intermediates of the PPP may provide new insights into metabolic defects underlying the accumulating polyols.
Huck
Jojanneke H J
JH
Department of Pediatrics, VU University Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
Struys
Eduard A
EA
Verhoeven
Nanda M
NM
Jakobs
Cornelis
C
van der Knaap
Marjo S
MS
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001800:Blood Specimen Collection; D002648:Child; D002675:Child, Preschool; D002853:Chromatography, Liquid; D005636:Fructosephosphates / Q000097:blood; D019298:Glucose-6-Phosphate / Q000097:blood; D005986:Glyceraldehyde 3-Phosphate / Q000097:blood; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D013058:Mass Spectrometry; D008875:Middle Aged; D010427:Pentose Phosphate Pathway; D010428:Pentosephosphates / Q000097:blood; D012267:Ribosemonophosphates / Q000097:blood; D012274:Ribulosephosphates / Q000097:blood; D012680:Sensitivity and Specificity; D013403:Sugar Phosphates / Q000097:blood; D014153:Transaldolase / Q000172:deficiency*
D005636:Fructosephosphates; D010428:Pentosephosphates; D012267:Ribosemonophosphates; D012274:Ribulosephosphates; D013403:Sugar Phosphates; D005986:Glyceraldehyde 3-Phosphate; C020495:sedoheptulose 7-phosphate; C031524:ribulose 5-phosphate; C031626:ribose-5-phosphate; D019298:Glucose-6-Phosphate; C026959:erythrose 4-phosphate; C031625:xylulose-5-phosphate; C027618:fructose-6-phosphate; D014153:Transaldolase