Pierre-Olivier Hétu1, Marie-Eve Gingras, Bernard Vinet. 1. Department of Biochemistry, Hôpital Notre-Dame, Centre Hospitalier de l'Université de Montréal, Montréal, Québec, Canada. pierre-olivier.hetu.chum@ssss.gouv.qc.ca
Abstract
OBJECTIVES: To develop an isotope dilution liquid chromatography tandem mass spectrometry (LC-IDMS/MS) method for the standardization of serum creatinine. DESIGN AND METHODS: Supernatants obtained by protein precipitation were injected into a LC-MS/MS. Chromatography was performed on a cation exchanger pre-column in normal phase isocratic mode. Creatinine and creatinine-D(3) were quantified using ion transitions of m/z 114-->44 and 117-->47, respectively. RESULTS: The method was calibrated with the NIST Standard Reference Material 914a and was found to be exact in analyzing the certified reference material SRM 967 from NIST (97.1+/-0.9% and 102.1+/-0.9% of target value for levels 1 and 2, respectively) and by calculating recuperation of spiked creatinine in 10 different patient samples (103.6+/-4.1%). Intra-assay imprecision was 0.9% at both 64.6 and 354 micromol/L creatinine, while inter-assay imprecisions were 1.9% and 1.8%. Absence of ion suppression was confirmed by spiking experiments. The method was shown to be free of carryover. A good correlation was obtained between the LC-MS/MS method and a Jaffe method run on an automated analyzer (r=0.999). CONCLUSIONS: We have developed a fast and simple method for the quantification of serum creatinine by isotope dilution tandem mass spectrometry (IDMS/MS) and we propose that this method can be used as a reference method by laboratories that wish to validate their serum creatinine automated assay. Published by Elsevier Inc.
OBJECTIVES: To develop an isotope dilution liquid chromatography tandem mass spectrometry (LC-IDMS/MS) method for the standardization of serum creatinine. DESIGN AND METHODS: Supernatants obtained by protein precipitation were injected into a LC-MS/MS. Chromatography was performed on a cation exchanger pre-column in normal phase isocratic mode. Creatinine and creatinine-D(3) were quantified using ion transitions of m/z 114-->44 and 117-->47, respectively. RESULTS: The method was calibrated with the NIST Standard Reference Material 914a and was found to be exact in analyzing the certified reference material SRM 967 from NIST (97.1+/-0.9% and 102.1+/-0.9% of target value for levels 1 and 2, respectively) and by calculating recuperation of spiked creatinine in 10 different patient samples (103.6+/-4.1%). Intra-assay imprecision was 0.9% at both 64.6 and 354 micromol/L creatinine, while inter-assay imprecisions were 1.9% and 1.8%. Absence of ion suppression was confirmed by spiking experiments. The method was shown to be free of carryover. A good correlation was obtained between the LC-MS/MS method and a Jaffe method run on an automated analyzer (r=0.999). CONCLUSIONS: We have developed a fast and simple method for the quantification of serum creatinine by isotope dilution tandem mass spectrometry (IDMS/MS) and we propose that this method can be used as a reference method by laboratories that wish to validate their serum creatinine automated assay. Published by Elsevier Inc.
Authors: Dennis R Koop; Lisa A Bleyle; Myrna Munar; Ganesh Cherala; Amira Al-Uzri Journal: J Chromatogr B Analyt Technol Biomed Life Sci Date: 2013-03-04 Impact factor: 3.205