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Development and Validation of Primary Method for the Determination of Glucose in Human Serum by Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry and Comparison with Field Methods

  • Lee, Hwa Shim (Center for Bioanalysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science) ;
  • Lee, Jong Man (Center for Bioanalysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science) ;
  • Park, Sang Ryoul (Center for Bioanalysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science) ;
  • Lee, Je Hoon (Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea) ;
  • Kim, Yong Goo (Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea)
  • Received : 2012.11.16
  • Accepted : 2013.03.13
  • Published : 2013.06.20

Abstract

Glucose is a common medical analyte measuring in human serum or blood samples. The development of a primary method is necessary for the establishment of traceability in measurements. We have developed an isotope dilution liquid chromatography tandem mass spectrometry as a primary method for the measurement of glucose in human serum. Glucose and glucose-$^{13}C_6$ in sample were ionized in ESI negative mode and monitored at mass transfers of m/z 179/89 and 185/92 in MRM, respectively. Glucose was separated on $NH_2P$-50 2D column, and the mobile phase was 20 mM $NH_4OAc$ in 30% acetonitrile/70% water. Verification of this method was performed by the comparison with NIST SRMs. Our results agreed well with the SRM values. We have developed two levels of glucose serum certified reference material using this method and distributed them to the clinical laboratories in Korea as samples for proficiency testings. The expended uncertainty was about 1.2% on 95% confidence level. In proficiency testings, the results obtained from the clinical laboratories showed about 3.6% and 3.9% RSD to the certified values. Primary method can provide the traceability to the field laboratories through proficiency testings or certified reference materials.

Keywords

References

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