FT-IR Methodology for Quality Control of Arabinogalactan Protein (AGP) Extracted from Green Tea (Camellia sinensis)

Abstract

A rapid methodology of quality control was developed for arabinogalactan proteins (AGP) extracted and purified from green tea. Using the vectorial angle method and IR spectrum analysis, the 1200-800 cm(-1) region in second-derivative IR spectra was determined as the key fingerprinting region of green tea AGP, with the 1090-900 cm(-1) region reflecting their conservative and common characteristics. In fact, the key monosaccharides, galactose (Gal) and arabinose (Ara), were shown to have intense peaks at about 1075 and 1045 cm(-1), respectively, and uronic acids at about 1018 cm(-1) in second-derivative IR spectra. The variable region was identified to be at about 1134-1094 and 900-819 cm(-1) and was probably due to compositional and structural differences between AGPs. The constructed methodology was tested on green tea AGP extracted by three treatments and purified to apparent homogeneity as water-extracted Camellia sinensis AGP (CSW-AGP), pectinase-extracted C. sinensis AGP (CSP-AGP), and trypsin-extracted C. sinensis AGP (CST-AGP) with an Ara/Gal ratio of 1.37, 1.57, and 1.82, respectively. Regarding in vitro antioxidant activity, the AGPs (CSW-AGP and CST-AGP) with higher similarity (closer cos theta values calculated for second-derivative IR spectra) exhibited a similar ability of chelating ferrous ions and had a similar capability for scavenging hydroxyl radicals. In conclusion, the combination of second-derivative IR spectrum analysis and the vectorial angle method has allowed a successful characterization of green tea AGPs and was shown to be suitable for their compositional and activity discrimination and rapid quality evaluation.