Comprehensive Two-Dimensional GC×GC-MS Method for Advanced Sugar Profiling in Human Biofluids

The composition of sugar compounds in human biofluids can be affected by several factors, such as diet, health, demographic background, and lifestyle. The accurate quantification of this profile allows for the identification and application of biomarkers that are connected to health and nutrition and provides useful insights into the mechanistic background of sugar metabolism. Existing methods typically quantify only a few sugar compounds simultaneously, however, thus constraining full assessment of the sugar profile. In response to this limitation, researchers at the Department of Safety and Quality of Fruit and Vegetables at the Max Rubner Institute (Karlsruhe, Germany) have proposeda comprehensive two-dimensional gas chromatography (GC×GC) coupled with a mass spectrometry-based detector sugaromics method applying a nonpolar-medium polar column setup. A paper based on their efforts was recently published in Analytical Chemistry.¹

The sugar profile of human biofluids is highly complex; up to 84 different sugar compounds can be detected simultaneously in urine and blood.^2-4The authors of the study¹ report that, despite the complex influence of diet, disease, and lifestyle on sugar profiles, surprisingly few analytical methods are currently able to quantify even ten or more sugar compounds simultaneously.

The researchers describe the method they developed as a combination of a targeted and untargeted approach to absolutely quantify various sugars in urine (n = 40) and serum (n = 36) and simultaneously identify untargeted sugar compounds in urine (n = 35) and serum (n = 22). The method was evaluated through a fit-for-purpose validation using the guidelines of the U.S. Food and Drug Administration and of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use as guiding principles. Most sugars subjected to the quantification demonstrated satisfactory validity parameters in terms of linearity, lower and upper limit of quantification, precision, accuracy, and carryover. In addition, the validated method was applied to human urine and serum samples (n = 40) and indicated quantifiable analyte concentrations within the expected range. In this respect, for healthy adults, absolute concentrations of seven sugars in urine and 14 in serum were reported for the first time.¹

The authors of the study write that the method “paves the way for application of the sugaromics method to investigate human nutrition and health and to assess metabolic pathways of sugars in the future.” They recommend that future work target ribonic and xylonic acid separation in urine and of other coeluting sugar compounds in both matrices.¹

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References

1. Streitenberger, E. E.; Egert, B.; Frommherz, L. et al. A Sugaromics Method for Combined Targeted and Untargeted Sugar Profiling: Fit-for-Purpose Validation of a Quantitative GC × GC-MS Approach. Anal Chem. 2026.DOI: 10.1021/acs.analchem.5c04230
2. Mack, C. I. Identification of Markers for Dietary Intake and Health Status by GC-MS Based Metabolite Profiling Approaches. Doctoral Thesis, Universität Hohenheim, 2020. https://hohpublica.uni-hohenheim.de/handle/123456789/6499.
3. Mack, C. I.; Ferrario, P. G.; Weinert, C. H. et al. Exploring the Diversity of Sugar Compounds in Healthy, Prediabetic, and Diabetic Volunteers. Mol Nutr Food Res. 2020, 64 (9), e1901190. DOI: 10.1002/mnfr.201901190
4. Mack, C. I.; Weinert, C. H.; Egert, B. et al. The Complex Human Urinary Sugar Profile: Determinants Revealed in the Cross-Sectional KarMeN Study. Am J Clin Nutr. 2018, 108 (3), 502-516. DOI: 10.1093/ajcn/nqy131