- P-ISSN 1225-0163
- E-ISSN 2288-8985
- P-ISSN1225-0163
- E-ISSN2288-8985
- SCOPUS, ESCI, KCI
Article Detail
Home > Article Detail
Quantitative determination of inosine 5’-monophosphate dehydrogenase activity in human peripheral blood mononuclear cells by ion-pair reversed-phase high-performance liquid chromatography
Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
2010, v.23 no.6, pp.531-536
https://doi.org/10.5806/AST.2010.23.6.531
&
(2010). Quantitative determination of inosine 5’-monophosphate dehydrogenase activity in human peripheral blood mononuclear cells by ion-pair reversed-phase high-performance liquid chromatography. Analytical Science and Technology, 23(6), 531-536, https://doi.org/10.5806/AST.2010.23.6.531
- Downloaded
- Viewed
Abstract
A quantitative analytical method has been established for the measurement of inosine 5’-monophosphate dehydrogenase (IMPDH) activity in human peripheral blood mononuclear cells (PBMCs) by ion-pair reversed-phase high performance liquid chromatography equipped with ultraviolet detection (HPLC/UV). IMPDH is a β-nicotinamide adenine dinucleotide hydrate (NAD+)-dependent dehydrogenase in which the enzyme converts inosine 5’-monophosphate (IMP) into xanthosine 5’-monophosphate (XMP). Its activity was measured by quantifying a HPLC chromatogram corresponding to XMP produced during the incubation of lysed PBMCs with IMP as a substrate and NAD+ as a coenzyme. XMP produced was detected at a wavelength of 260 nm. The mobile phase was composed of a mixture of 37 mM potassium dihydrogen phosphate containing 7 mM tetra-n-butylammonium hydrogen sulfate adjusted to pH 5.5 and methanol (85:15, v/v) with a flow rate of 1 mL/min. The calibration curve was linear (r2=0.999999) in the range of 0.2-50.0 μM and the limit of quantification (LOQ) was 0.2 μM. The intra- and inter-day precisions were between 0.88-1.47% and 0.85-5.24%,respectively. The intra- and inter-day accuracies were between 98.74-99.99% and 99.95-101.65%, respectively. IMPDH activity in 11 Korean healthy volunteers ranged from 18.29 to 36.60 nmol/h/mg protein (mean = 27.70± 6.28 nmol/h/mg protein).
- keywords
- IMP, XMP, IMPDH activity, PBMCs, HPLC
Reference
1
1. W. Wang and L. Hedstrom, Biochemistry, 36(28), 8479-8483(1997).
2
2. A. C. Allison and E. M. Eugui, Immunopharmacology, 47(2-3), 85-118(2000).
3
3. E. M. Eugui and A. C. Allison, Ann. N. Y. Acad. Sci, 685, 309-329(1993).
4
4. M. A. Fleming, S. P. Chambers, P. R. Connelly, E. Nimmesgern, F. Fox, F. J. Bruzzese, S. T. Hoe, J. R. Fulghum, D. J. Livingston, C. M. Stuver, M. D. Sintchak, K. P. Wilson and J. A. Thomson, Biochemistry, 35(22), 6990-6997(1996).
5
5. A. C. Allison, T. Hovi, R. W. Watts and A. D. Webster, Lancet, 2(7946), 1179-1183 (1975).
6
6. A. C. Allison and E. M. Eugui, Springer Semin. Immunopathol., 14(4), 353-380(1993).
7
7. Y. Natsumeda, S. Ohno, H. Kawasaki, Y. Konno, G. Weber and K. Suzuki, J. Biol. Chem., 265(9), 5292- 5295(1990).
8
8. S. F. Carr, E. Papp, J. C. Wu and Y. Natsumeda, J. Biol. Chem., 268(36), 27286-27290(1993).
9
9. Y. Konno, Y. Natsumeda, M. Nagai, Y. Yamaji, S. Ohno, K. Suzuki and G. Weber, The J. Biol. Chem., 266(1), 506-509(1991).
10
10. L. R. Chiarelli, M. Molinaro, C. Libetta, C. Tinelli, L. Cosmai, G. Valentini, A. Dal Canton and M. Regazzi, Br. J. Clin. Pharmacol, 69(1), 38-50(2010).
11
11. E. Jacqz-Aigrain, E. Bessa, Y. Medard, Y. Mircheva and E. Vilmer, Br. J. Clin. Pharmacol., 38(1), 1-8(1994).
12
12. T. Furuchi, S. Kosugi, K. Ohno, T. Egawa, M. Sekine, M. Katane and H. Homma, Anal. Biochem., 384(2), 207-212(2009).
13
13. L. J. Langman, D. F. LeGatt and R. W. Yatscoff, Clin. Chem., 41(2), 295-299(1995).
14
14. P. Glander, K. P. Braun, P. Hambach, S. Bauer, I. Mai, I. Roots, J, Waiser, L. Fritsche, H. H. Neumayer, K. Budde, Clin. Biochem., 34(7), 543-549(2001).
15
15. T. Maiguma, T. Yosida, K. Otsubo, Y. Okabe, A. Sugitani, M. Tanaka, R. Oishi and D. Teshima, J. Clin. Pharm. Ther., 35(1), 79-85(2010).
16
16. U.S. Department of Health and Human Services, Food and Drug Administration, Guidance for Industry: Bioanalytical Method Validation (2001).
Other articles from this issue
- Dissolution of vanadium pentoxide using microwave digestion system for determination of vanadium by ICP-AES
- A study on the analysis of uranium isotopes in environmental samples using a kinetic phosphorescence analyzer
- Accurate determination of chloramphenicol in meat by isotope dilution liquid chromatography mass spectrometry (ID-LC/MS)
- Quantitative determination of inosine 5’-monophosphate dehydrogenase activity in human peripheral blood mononuclear cells by ion-pair reversed-phase high-performance liquid chromatography
- Bioconjugation by dual heterobifunctional coupling method:Use of the conjugates for the detection of dopamine
- Simultaneous analysis of ethylene glycol and glycolic acid in bio-specimens by GC/MS
- Optimization of analytical conditions for the determination of nitrosamines in chlorinated tap water by high performance liquid chromatography
- Reduction of perchlorate in aqueous solution using zero valence iron stabilized with alginate bead
- Sorption of aqueous uranium(VI) ion onto a cation-exchangeable K-birnessite colloid
- Simultaneous analysis and occurrences of six pharmaceuticals in surface water by LC/ESI-MS/MS
- Antitumor agents bound to silica nanoparticles : potential technology for the remediation of malignant tumors
- Analytical method validation of oxiracetam using HPLC
- Exposure assessment of dioxins through foods
- A study on shape changes of hangul signature under the influence of alcohol
- Analysis of heavy metal in polymer materials by combustion ICP-AES Method
- more
Recommanded Articles
This website recommended the use the Chrome browser for journal website.