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Separation and identification of chlorogenic acids using the HPLC method

Chlorogenic acids (CGAs) are a group of polyphenolic compounds distributed in different plants. CGAs are a family of different esters formed between the (-)-quinic acid and certain trans-cinnamic acids (caffeic, ferulic). In this study, the separation and identification of nine different CGAs isomers: 5-O- caffeoylquinic acid (5-CQA), 4-O-caffeoylquinic acid (4-CQA), 3-O-caffeoylquinic acid (3-CQA), 3, 5-O-dicaffeoylquinic acid (3, 5-diCQA), 4, 5-O- dicaffeoylquinic acid (4, 5-diCQA), 3, 4- O- dicaffeoylquinic acid (3, 4-diCQA) and 5-O- feruloylquinic acid (5-FQA), 4-O-feruloylquinic acid (4-FQA), and 3-O-feruloylquinic acid (3- FQA) was performed in a model solution using high performance liquid chromatography (HPLC). The HPLC analysis was carried out on the HPLC system (Agilent) with a PDA detector using the C18 column and two mobile phases, 0.1% phosphoric acid in water, and 100% methanol. UV/Vis spectra were recorded in the wavelength range from 200 nm to 600 nm. Chlorogenic acids have shown the maximum absorbance at the wavelength of 320 nm. The order of eluation of the individual chlorogenic acids is consistent with the literature data. All investigated chlorogenic acids are well separated, except the 3, 4 – and 3, 5-diCQA (the retention times are very close). The method has shown linearity in the range of concentrations from 20 mg/L to 300 mg/L, limit of detection (LOD) was 1.7 · 10-5mol/L and the limit of quantification (LOQ) was 5.7 · 10-5mol/L. This method could be applied in the analysis of chlorogenic acids in various beverages.

chlorogenic acids, HPLC, validation

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