Quantitation of carbohydrate monomers and dimers by liquid chromatography coupled with high-resolution mass spectrometry.

As remnants of plant wastes or plant secretions, carbohydrates are widely found in various environmental matrices. Carbohydrate-containing feedstocks represent important carbon sources for engineered bioproduction of commodity compounds. Routine monitoring and quantitation of heterogenous carbohydrate mixtures requires fast, accurate, and precise analytical methods. Here we present two methods to quantify carbohydrates mixtures by coupling hydrophilic interaction liquid chromatography with electrospray ionization high-resolution mass spectrometry. Method 1 was optimized for eleven different carbohydrates: three pentoses (ribose, arabinose, xylose), three hexoses (glucose, fructose, mannose), and five dimers (sucrose, cellobiose, maltose, trehalose, lactose). Method 1 can monitor these carbohydrates simultaneously, except in the case of co-elution of xylose/arabinose and lactose/maltose/cellobiose peaks. Using the same stationary and mobile phases as in Method 1, Method 2 was developed to separate glucose and galactose, which were indistinguishable in Method 1. Both methods have low limits of detection (0.019-0.40 μM) and quantification (0.090-1.3 μM), good precision (2.4-13%) except sucrose (18%), and low mass error (0.0-2.4 ppm). Method 1 was robust at analyzing high ionic strength solutions, but a moderate matrix effect was observed. Finally, we apply Method 1 to track concurrently the extracellular depletion of five carbohydrates (xylose, glucose, fructose, mannose, and maltose) by Pseudomonas protegens Pf-5, a biotechnologically-important soil bacterial species.