Quantitative ¹H-NMR Spectroscopy for Profiling Primary Metabolites in Mulberry Leaves.

The primary metabolites in aqueous extract of mulberry (Morus alba L.) leaves were characterized by using proton nuclear magnetic resonance (¹H-NMR) spectroscopy. With the convenience of resonance assignment, GABA together with the other 10 primary metabolites was simultaneously identified and quantified in one ¹H-NMR spectrum. In this study, external calibration curves for metabolites were employed to calculate the concentrations of interests. The proposed quantitative approach was demonstrated with good linearity (r² ranged in the interval of 0.9965-0.9999), precision, repeatability, stability (RSD values in the ranges of 0.35-4.89%, 0.77-7.13% and 0.28-2.33%, respectively) and accuracy (recovery rates from 89.2% to 118.5%). The established ¹H-NMR method was then successfully applied to quantify 11 primary metabolites in mulberry leaves from different geographical regions within a rapid analysis time and a simple sample preparation procedure.