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Analysis of glycolysis metabolites by capillary zone electrophoresis with indirect UV detection.
J Capillary Electrophor. 1996 May-Jun; 3(3):155-63.JC

Abstract

The glycolysis metabolites glucose 6-phosphate (G6-P), fructose 6-phosphate (F6-P), fructose 1,6-bisphosphate (F1,6-BP), fructose 2,6-bisphosphate (F2,6-BP), glyceraldehyde phosphate (GAP), dihydroxyacetone phosphate (DHAP), phosphoenolpyruvate (PEP), pyruvate, and lactate were analyzed by capillary zone electrophoresis (CZE) with indirect UV detection. The chromophores phthalic acid, sorbic acid, and 4-hydroxybenzoic acid were studied as background electrolytes. Both detection sensitivity and resolution were found to depend on the pH and the concentration of the carrier electrolyte. Optimum separation and detection of the phosphate compounds were accomplished upon reversal of electroendosmotic flow (EOF) with OFM Anion-BT (Waters Corp., Milford, MA) at a concentration of 4-6 mM 4-hydroxybenzoic acid, pH 11.6-12.0, with the detection wave-length set at 280 nm. The highly alkaline pH allowed the successful separation of the isomers F6-P and G6-P, as well as F1,6-BP and F2,6-BP, respectively. The effect of sample ionic strength on the detection limits of G6-P, F6-P, F1,6-BP, and F2,6-BP was also investigated: These limits ranged from 1 to 3 microM in both low- and high-ionic-strength samples. However, high Mg2+ concentrations in the sample led to a progressive loss of resolution between F1,6-BP and F2,6-BP, unless the inlet reservoir was replenished with fresh electrolyte after every injection. Linearity of detection was observed over one to two orders of magnitude.

Authors+Show Affiliations

Shen P
Department of Chemistry, Stanford University, CA 94305-5080, USA.
Hauri D
No affiliation info available
Ross J
No affiliation info available
Oefner PJ
No affiliation info available

MeSH

Dihydroxyacetone PhosphateElectrophoresis, CapillaryFructosediphosphatesFructosephosphatesGlucose-6-PhosphateGlycerophosphatesGlycolysisIndicators and ReagentsLactatesPhosphoenolpyruvatePyruvatesReproducibility of ResultsSensitivity and SpecificitySpectrophotometry, UltravioletSugar Phosphates

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

9384748

Citation

Shen, P, et al. "Analysis of Glycolysis Metabolites By Capillary Zone Electrophoresis With Indirect UV Detection." Journal of Capillary Electrophoresis, vol. 3, no. 3, 1996, pp. 155-63.
Shen P, Hauri D, Ross J, et al. Analysis of glycolysis metabolites by capillary zone electrophoresis with indirect UV detection. J Capillary Electrophor. 1996;3(3):155-63.
Shen, P., Hauri, D., Ross, J., & Oefner, P. J. (1996). Analysis of glycolysis metabolites by capillary zone electrophoresis with indirect UV detection. Journal of Capillary Electrophoresis, 3(3), 155-63.
Shen P, et al. Analysis of Glycolysis Metabolites By Capillary Zone Electrophoresis With Indirect UV Detection. J Capillary Electrophor. 1996 May-Jun;3(3):155-63. PubMed PMID: 9384748.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analysis of glycolysis metabolites by capillary zone electrophoresis with indirect UV detection. AU - Shen,P, AU - Hauri,D, AU - Ross,J, AU - Oefner,P J, PY - 1996/5/1/pubmed PY - 1997/12/31/medline PY - 1996/5/1/entrez SP - 155 EP - 63 JF - Journal of capillary electrophoresis JO - J Capillary Electrophor VL - 3 IS - 3 N2 - The glycolysis metabolites glucose 6-phosphate (G6-P), fructose 6-phosphate (F6-P), fructose 1,6-bisphosphate (F1,6-BP), fructose 2,6-bisphosphate (F2,6-BP), glyceraldehyde phosphate (GAP), dihydroxyacetone phosphate (DHAP), phosphoenolpyruvate (PEP), pyruvate, and lactate were analyzed by capillary zone electrophoresis (CZE) with indirect UV detection. The chromophores phthalic acid, sorbic acid, and 4-hydroxybenzoic acid were studied as background electrolytes. Both detection sensitivity and resolution were found to depend on the pH and the concentration of the carrier electrolyte. Optimum separation and detection of the phosphate compounds were accomplished upon reversal of electroendosmotic flow (EOF) with OFM Anion-BT (Waters Corp., Milford, MA) at a concentration of 4-6 mM 4-hydroxybenzoic acid, pH 11.6-12.0, with the detection wave-length set at 280 nm. The highly alkaline pH allowed the successful separation of the isomers F6-P and G6-P, as well as F1,6-BP and F2,6-BP, respectively. The effect of sample ionic strength on the detection limits of G6-P, F6-P, F1,6-BP, and F2,6-BP was also investigated: These limits ranged from 1 to 3 microM in both low- and high-ionic-strength samples. However, high Mg2+ concentrations in the sample led to a progressive loss of resolution between F1,6-BP and F2,6-BP, unless the inlet reservoir was replenished with fresh electrolyte after every injection. Linearity of detection was observed over one to two orders of magnitude. SN - 1079-5383 UR - https://www.unboundmedicine.com/medline/citation/9384748/Analysis_of_glycolysis_metabolites_by_capillary_zone_electrophoresis_with_indirect_UV_detection_ L2 - https://antibodies.cancer.gov/detail/CPTC-CLIC1-1 DB - PRIME DP - Unbound Medicine ER -