LC-MS/MS quantitative analysis of reducing carbohydrates in soil solutions extracted from crop rhizospheres

A simple, sensitive, and specific analytical method has been developed for the quantitative determination of 15 reducing carbohydrates in the soil solution of crop rhizosphere. Reducing carbohydrates were derivatized with 1-phenyl-3-methyl-5-pyrazolone, separated by reversed-phase high-performance liquid chromatography and detected by electrospray ionization tandem mass spectrometry. Lower limits of quantitation of 2 ng/mL were achieved for all carbohydrates. Quantitation was performed using peak area ratios (analyte/internal standard) and a calibration curve spiked in water with glucose-d(2) as the internal standard. Calibration curves showed excellent linearity over the range 2-100 ng/mL (10-1,000 ng/mL for glucose). The method has been tested with quality control samples spiked in water and soil solution samples obtained from the rhizosphere of wheat and canola and has been found to provide accurate and precise results.