Measurement of glucose and fructose in clinical samples using gas chromatography/mass spectrometry

Abstract

Objective The impact of increased fructose consumption on carbohydrate metabolism is a topic of current interest, but determination of serum level has been hindered due to low concentration and interference from serum glucose. We are reporting a method for the quantification of glucose and fructose in clinical samples using gas chromatography/mass spectrometry (GC/MS). The accuracy and precision of GC/MS and an enzymatic assay were compared. Design and methods Mass spectrometry fragmentation patterns of methyloxime peracetate derivatized aldose and ketose were determined. Unique fragments for glucose and fructose were used for quantitative analysis using isotope labeled recovery standards. Results Methyloxime peracetate derivatives of glucose and fructose showed characteristic loss of acetate (M-60) or ketene (M-42) under chemical ionization (CI). Under electron impact (EI) ionization, a unique C1-C2 fragment of glucose was formed, while a C1-C3 fragment was formed from keto-hexoses. These unique fragments were used in the quantitative assay of glucose and fructose in clinical samples. In clinical samples, the GC/MS assay has a lower limit of detection than that of the enzymatic assay. In plasma samples from patients evaluated for diabetes the average serum glucose and fructose were 6.19±2.72 mM and 46± 25.22 μM. Fructose concentrations in many of these samples were below the limit of detection of the enzymatic method. Conclusion Derivatization of aldose and ketose monosaccharides to their respective O-methyloxime acetates for GC/MS analysis is a facile method for determination of serum/plasma glucose and fructose samples.

Fructose metabolism and metabolic disease

“…Fructose concentrations in peripheral plasma are typically about 0.04 mM, can acutely increase 10-fold after fructose consumption, and return to fasting levels within 2 hours (39)(40)(41). This rapid clearance is mediated in large part by efficient extraction by the liver.…”

Quality control of sweet medicines based on gas chromatography-mass spectrometry

“…The derivatization of aldose and alditols to aldononitrile acetates was successfully archived single peaks and quantification analysis of carbohydrates in Ganoderma (38). However, the validity of quantitative analysis using these derivatives also has the drawback that they cannot be applied for analysis of ketoses (14,31). Actually, when aldoses and ketoses exist simultaneously in the matrix, samples can go through oximation first and subsequent derivatization by acetylation and silylation, respectively, and finally converted to aldononitrile acetates and TMSO respectively.…”

“…Derivatization of aldose and ketose to their respective O-methyloxime acetates (oximation by methoxiamine hydrochloride and acetylation by acetic anhydride) for GC-MS analysis is also a facile method for the determination when aldoses and ketoses simultaneously existed, and more stable and sensitive than TMSO. Moreover, O-methyloxime acetates derivatives of glucose and fructose showed characteristic fragments both in chemical ionization (CI) and electronic ionizaton (EI) mode of mass spectrometry (31).…”

“…All of these oximes derivatives produce two peaks corresponding to the syn (E) and anti (Z) forms per reducing sugar and applicable to aldoses, ketoses and alditols (except aldononitrile acetates). As relative simple chromatograms are achieved, these derivatives have also been adopted for carbohydrate analysis in complex mixtures (30)(31)(32)(33). The comparison of single-step derivatization trimethylsilylation and two-step approaches including ethoximation-trimethylsilylation (EO-TMS), ethoximation-trifluoroacetylation (EO-TFA), benzoximation-trimethylsilylation (BO-TMS) and benzoximation-trifluoroacetylation (BO-TFA) have been comprehensively studied for derivatization of aldoses and ketoses with regard to chromatographic characteristics.…”

Optimization of Differential pH Sensors Device Operation Conditions to Be Used in Quantification of Low Glucose Concentration

“…Determining glucose concentration for many applications was done by using different methods such as gas chromatography-mass spectrometry, HPLC, electrochemical and enzymatic assay (Mosca et al, 1981;Xie et al, 2009;Diessel et al, 2004;Ciantar et al, 2002;Boduroglu et al, 2005;Barrado et al, 1999;Mori et al, 1999;Wahjudi et al, 2010).…”