A novel ion-exclusion chromatography–mass spectrometry method to measure concentrations and cycling rates of carbohydrates and amino sugars in freshwaters

• Novel HPLC–MS method for carbohydrate and amino sugar analysis in freshwaters.
• Rapid and efficient determination without derivatization or sample pre-concentration.
• Implementation of stable isotope tracers to study compound biogeochemical cycling.
• Glucose and N-acetyl-glucosamine showed different in situ dynamics in Lake Zurich.

The concentrations of free neutral carbohydrates and amino sugars were determined in freshwater samples of distinct matrix complexity, including meso-, eu- and dystrophic lakes and ponds, using high-performance ion-exclusion chromatography (HPIEC) coupled to mass spectrometry (MS). In contrast to other methods, our approach allowed the quantification of free neutral carbohydrates and amino sugars at low nM concentrations without derivatization, de-salting or pre-concentration. New sample preparation procedures were applied prior to injection employing syringe and hollow fiber filtration. Analytes were separated on a strong cation exchange resin under 100% aqueous conditions using 0.1% formic acid as a mobile phase. To minimize background noise in MS, analytes were detected in a multiple reaction monitoring scan mode with double ion filtering. Detection limits of carbohydrates and amino sugars ranged between 0.2 and 2 nM at a signal-to-noise ratio >5. Error ranged between 1 and 12% at 0.5–500 nM levels. Using a stable isotope dilution approach, both the utilization and recycling of glucose in Lake Zurich was observed. In contrast, N-acetyl-glucosamine was equally rapidly consumed but there was no visible de novo production. The simple and rapid sample preparation makes our protocol suitable for routine analyses of organic compounds in freshwater samples. Application of stable isotope tracers along with accurate measures of carbohydrate and amino sugar concentrations enables novel insights into the compound in situ dynamics.