A Fast Determination of Chlorophylls in Barley Grass Juice Powder Using HPLC Fused-Core Column Technology and HPTLC
Abstract
This work presents a novel approach to qualitative and quantitative evaluation of chlorophyll content in food supplement using simple and fast separation techniques (high performance liquid chromatography (HPLC) and high performance thin layer chromatography (HPTLC)). For the first time, an AscentisA (R) Express C-18 HPLC column (100 x 4.6 mm; 2.7 mu m), based on fused-core particles, was utilised for the analysis of chlorophyll a and chlorophyll b in green young barley grass juice powder extracts.
The mobile phase was composed of methanol, water and ethyl acetate of 40:10:50 (v/v/v) at a flow rate of 1.5 mL/min. Chlorophylls were separated in time less than 6 min.
The extraction procedure was optimised by using of 90 % of acetone in water, and the method was validated. The method precision was found in the range of 1.15-2.85 %, and the method recovery was established at three concentration levels.
Calibration graphs were linear in the range of 0.75-25.5 mg/L, with correlation coefficients r (2) = 0.99958 for chlorophyll a and r (2) = 0.99961 for chlorophyll b. The comparative HPTLC fingerprint analysis of three samples from different continents showed only slight qualitative differences between the particular samples.
The results demonstrated that the proposed methods can be used as fast and powerful techniques for the characterisation of barley juice extracts in quality control.