A high-performance thin-layer chromatographic method for chlorogenic acid and hyperoside determination from berry extracts

The present study investigates a quantitative high performance thin layer chromatographic (HPTLC) method for chlorogenic acid and hyperoside determination from berry extracts. Chlorogenic acid and hyperoside were extracted using a methanol and water mixture and separated on a HPTLC 60F254 silica gel plates with ethyl acetate, acetic acid, formic acid and water 10:1.1:1.1:2.3 (v/v/v/v) as mobile phase within 35 minutes. After chromatography, the plates were sprayed with natural products (polyethylene glycol reagent (NP/PEG)). Densitometric analysis of these compounds was performed via fluorescence, under UV light at λ = 366 nm. The Rf of chlorogenic acid and hyperoside was 0.74 and respectively 0.85. Calibration data revealed good linear and polynomial relationships (r = 0.99996 and 0.99999) between peak height area and chlorogenic acid, respectively hyperoside concentration. Based on the results obtained in this study, the investigated method, can be used for rapid routine analysis of chlorogenic acid and hyperoside.