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Determination of serum uric acid using high-performance liquid chromatography (HPLC)/isotope dilution mass spectrometry (ID-MS) as a candidate reference method.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Oct 01; 857(2):287-95.JC

Abstract

Uric acid is an important diagnostic marker of catabolism of the purine nucleosides, and accurate measurements of serum uric acid are necessary for proper diagnosis of gout or renal disease appearance. A candidate reference method involving isotope dilution coupled with liquid chromatography/mass spectrometry (LC/MS) has been described. An isotopically labeled internal standard, [1,3-(15)N(2)] uric acid, was added to serum, followed by equilibration and protein removal clean up to prepare samples for liquid chromatography/mass spectrometry electrospray ionization (LC/MS-ESI) analyses. (M-H)(-) ions at m/z 167 and 169 for uric acid and its labeled internal standard were monitored for LC/MS. The accuracy of the measurement was evaluated by a comparison of results of this candidate reference method on lyophilized human serum reference materials for uric acid (Standard Reference Materials SRM909b) with the certified values determined by gas chromatography/mass spectrometry reference methods and by a recovery study for the added uric acid. The method performed well against the established reference method of ion-exchange followed by derivatization isotope dilution (ID) gas chromatography mass spectrometry (ID-GC/MS). The results of this method for uric acid agreed well with the certified values and were within 0.10%. The amounts of uric acid recovered and added were in good agreement for the three concentrations. This method was applied to determine uric acid in samples of frozen serum pools. Excellent precision was obtained with within-set CVs of 0.08-0.18% and between-set CVs of 0.02-0.07% for LC/MS analyses. Liquid chromatography/tandem mass spectrometry electrospray ionization (LC/MS/MS-ESI) analysis was also performed. The LC/MS and LC/MS/MS results were in very good agreement (within 0.14%). This LC/MS method, which demonstrates good accuracy and precision, and is in the speed of analysis without the need for a derivatization stage, qualifies as a candidate reference method. This method can be used as an alternative reference method to provide an accuracy base to which the routine methods can be compared.

Authors+Show Affiliations

National Institute of Metrology, National Research Center for CRMs, Beijing 100013, China. xhdai.75@iccas.ac.cnNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

17704012

Citation

Dai, Xinhua, et al. "Determination of Serum Uric Acid Using High-performance Liquid Chromatography (HPLC)/isotope Dilution Mass Spectrometry (ID-MS) as a Candidate Reference Method." Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, vol. 857, no. 2, 2007, pp. 287-95.
Dai X, Fang X, Zhang C, et al. Determination of serum uric acid using high-performance liquid chromatography (HPLC)/isotope dilution mass spectrometry (ID-MS) as a candidate reference method. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;857(2):287-95.
Dai, X., Fang, X., Zhang, C., Xu, R., & Xu, B. (2007). Determination of serum uric acid using high-performance liquid chromatography (HPLC)/isotope dilution mass spectrometry (ID-MS) as a candidate reference method. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 857(2), 287-95.
Dai X, et al. Determination of Serum Uric Acid Using High-performance Liquid Chromatography (HPLC)/isotope Dilution Mass Spectrometry (ID-MS) as a Candidate Reference Method. J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Oct 1;857(2):287-95. PubMed PMID: 17704012.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Determination of serum uric acid using high-performance liquid chromatography (HPLC)/isotope dilution mass spectrometry (ID-MS) as a candidate reference method. AU - Dai,Xinhua, AU - Fang,Xiang, AU - Zhang,Chunmei, AU - Xu,Ruifeng, AU - Xu,Bei, Y1 - 2007/08/02/ PY - 2007/02/06/received PY - 2007/07/18/revised PY - 2007/07/23/accepted PY - 2007/8/21/pubmed PY - 2007/12/7/medline PY - 2007/8/21/entrez SP - 287 EP - 95 JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences JO - J Chromatogr B Analyt Technol Biomed Life Sci VL - 857 IS - 2 N2 - Uric acid is an important diagnostic marker of catabolism of the purine nucleosides, and accurate measurements of serum uric acid are necessary for proper diagnosis of gout or renal disease appearance. A candidate reference method involving isotope dilution coupled with liquid chromatography/mass spectrometry (LC/MS) has been described. An isotopically labeled internal standard, [1,3-(15)N(2)] uric acid, was added to serum, followed by equilibration and protein removal clean up to prepare samples for liquid chromatography/mass spectrometry electrospray ionization (LC/MS-ESI) analyses. (M-H)(-) ions at m/z 167 and 169 for uric acid and its labeled internal standard were monitored for LC/MS. The accuracy of the measurement was evaluated by a comparison of results of this candidate reference method on lyophilized human serum reference materials for uric acid (Standard Reference Materials SRM909b) with the certified values determined by gas chromatography/mass spectrometry reference methods and by a recovery study for the added uric acid. The method performed well against the established reference method of ion-exchange followed by derivatization isotope dilution (ID) gas chromatography mass spectrometry (ID-GC/MS). The results of this method for uric acid agreed well with the certified values and were within 0.10%. The amounts of uric acid recovered and added were in good agreement for the three concentrations. This method was applied to determine uric acid in samples of frozen serum pools. Excellent precision was obtained with within-set CVs of 0.08-0.18% and between-set CVs of 0.02-0.07% for LC/MS analyses. Liquid chromatography/tandem mass spectrometry electrospray ionization (LC/MS/MS-ESI) analysis was also performed. The LC/MS and LC/MS/MS results were in very good agreement (within 0.14%). This LC/MS method, which demonstrates good accuracy and precision, and is in the speed of analysis without the need for a derivatization stage, qualifies as a candidate reference method. This method can be used as an alternative reference method to provide an accuracy base to which the routine methods can be compared. SN - 1570-0232 UR - https://www.unboundmedicine.com/medline/citation/17704012/Determination_of_serum_uric_acid_using_high_performance_liquid_chromatography__HPLC_/isotope_dilution_mass_spectrometry__ID_MS__as_a_candidate_reference_method_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1570-0232(07)00545-4 DB - PRIME DP - Unbound Medicine ER -