Simultaneous determination of chlorogenic acid, caffeic acid and caffeine in hydroalcoholic and aqueous extracts of Ilex paraguariensis by HPLC and correlation with antioxidant capacity of the extracts by DPPH· reduction

Abstract

A new high performance liquid chromatographic method has been established for simultaneous determination of chlorogenic acid, caffeic acid and caffeine in hydroalcoholic and aqueous extracts ofIlex paraguariensis. Analytical curves showed good linear regression in the concentration ranges 0.49-7.8 μg/mL for chlorogenic acid, 0.25-3.9 μg/mL for caffeic acid and 0.244-7.8 μg/mL for caffeine. Reduction of the DPPH· radical was used to determine the antioxidant capacity of the extracts. Our method for the simultaneous determination of chlorogenic acid, caffeic acid and caffeine was highly sensitive, having lower detection and quantitation limits than other papers that used similar methodology. INTRODUCTIONA free radical is any chemical species capable of independent existence that contains one or more unpaired electrons (Halliwel, Gutteridge, 1999). The role of freeradical reactions in biology has become an area of intense interest. It is generally accepted that free radicals play an important role in the development of tissue damage and pathological events in living organisms (De Groot, Noll, 1987;Schinella et al., 2000). The consumption of plant foods is associated with a reduced risk of chronic diseases (cancer and cardiovascular), in part because of substances with antioxidant capacity, and those that modulate enzyme activity and gene expression (Day et al., 2004).Antioxidative properties have been found in leaf infusions of yerba mate (Ilex paraguariensis) (Chandra, Mejia, 2004). Mate is a popular tea in South America, where it is frequently drunk instead of coffee as a stimulating drink. It is greenish in color and contains caffeine, tannins and several vitamins such as B1 (thiamine), B2 (riboflavin), B5 (pantothenic acid), C, E, β-carotene, sucrose, fructose, folic acid, trigonelline, choline and many polyphenolic compounds (flavonoids and chlorogenic acid) (Gugliucci, Stahl, 1995). Much experimental work (in vivo and in vitro) has been performed to evaluate the antioxidant capacity of yerba mate. The leaf extract of I. paraguariensis was shown to be absorbed and to be active in vivo when healthy patients drank it, and one hour later blood was collected and the LDL content analysed in relation to lipid peroxidation (Gugliucci, 1996). The Ilex paraguariensis aqueous / ascorbate system and that induced by the CCl 4 /NADPH system, in both cases detected by the thiobarbituric acid assay (Schinella et al., 2000) For the simultaneous determination of CGA (chlorogenic acid) and CFA (caffeic acid) (Figure 1) by HPLC, various sets of chromatographic conditions have been used in recent work: in Li et al. (2004), the mobile phase was a mixture of methanol and 0.2 mol/L acetate buffer (pH 3.6) (15:85, v/v) flowing at 1.0 mL/min, the detection wavelength (λ) being set at 300 nm; in Li et al. (2005), methanol:water:acetic acid (19:81:1.5, v/v) was used, flowing at 1.0 mL/min, with λ at 240 nm, and in Pellati et al. (2005), aqueous phosphoric acid solution (0.1%) and acetonitrile (90:10 v/v), flowing at 1.5 mL/min, with the...

Influence of the addition of natural antioxidant from mate leaves (Ilex paraguariensis St. Hill) on the chemical, microbiological and sensory characteristics of different formulations of Prato cheese

“…Moure et al (2001) showed that the stability of polyphenol compounds during the extraction and dehydration is affected by chemical and enzymatic degradations, by the volatilization of compounds and mainly by the thermal decomposition. Rivelli et al (2007) Brasil 1996). Figure 1a presents the pH values of the different formulations tested in this work.…”

The influence of leaf age on methylxanthines, total phenolic content, and free radical scavenging capacity of Ilex paraguariensis aqueous extracts

“…Mate is used for its central nervous system stimulant properties, which are due to the presence of the methylxanthines caffeine and theobromine (Blumenthal and Brinckmann, 2000;Dermarderosian, 2001;Filip et al, 1998). Additionally, yerba-mate is also considered a functional food, because of its nutritional and medicinal properties, such as hypocholesterolemic, hepatoprotective, diuretic, and antioxidant properties (Bixby et al, 2005;Filip et al, 2000;Gugliucci and Stahl, 1995;Heck and De Mejia, 2007;Rivelli et al, 2007;Valerga et al, 2012), which can protect against the harmful effect of free radicals, thereby increasing the defense system of the organism. It can also help prevent atherosclerosis and coronary heart disease (Heck and De Mejia, 2007;Miranda et al, 2008;Puangpraphant and de Mejia, 2009).…”

Are coffee silverskin extracts safe for topical use? An in vitro and in vivo approach

“…The HPLC method used in this study to quantify HMF, CQA and caffeine in CS extracts was based on the work of Lemos et al (2010) and Rivelli et al (2007). The chromatographic analysis was per-formed in a HPLC integrated system (Jasco, Japan) equipped with a PU-980 pump and a Jasco AS-950 automatic sampler with a 20 µL loop.…”