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

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- 13C 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 2 P-50 2D column, and the mobile phase was 20 mM NH 4 OAc 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.

Glucose and glycerol concentrations and their tracer enrichment measurements using liquid chromatography tandem mass spectrometry

“…[22] However, glucose 13 C-enrichment in human plasma can also be measured using LC-IRMS [22] and glucose D 2enrichment can be measured using LC-MS. [6,14] Two LC-MS/MS methods to measure glucose concentration (not tracer enrichment) in underivatized human serum have recently been published. [23,24] However, characteristic for both methods is that three out of six 13 C-labeled carbon atoms of the internal standard were lost. Therefore, these methods are not suited to measure tracer enrichment in connection to concentration measurements.…”

Screening of Potential Stress Biomarkers in Sweat Associated with Sports Training

“…The tandem mass spectrometry conditions were first selected through bibliographic research [11,17,23,25,[27][28][29][30][31][32][33]. The analytical standard analyte of E, NE, Phe, Trp, Tyr, His, Lys and Asc was used to confirm the biomarker identification (these have certified content and purity that is used as a reference in the analysis).…”

Development of an SI-Traceable HPLC–Isotope Dilution Mass Spectrometry Method To Quantify β-Lactoglobulin in Milk Powders

“…It allows highly accurate and precise measurements of various substances and has been used widely since its development in the 1980s. − It has been used to detect and quantify drugs, hormones, pesticides and veterinary drug residues in serum, and other organic small molecules with high accuracy. With the development of the life science and biotechnology industries, IDMS methods have been further developed to quantify proteins, − nucleic acids, and other biological macromolecules. − Especially, IDMS has been successfully applied into analyses with milk powders and shows several advantages. Zhang et al developed an IDMS method to quantify12 mycotoxins in milk-based infant formula and foods.…”