Gema Baeza

Hypocholesterolaemic and antioxidant effects of yerba mate (Ilex paraguariensis) in high-cholesterol fed rats

OBJECTIVE To study the effect of mate (Ilex paraguariensis) on serum lipids and antioxidant status in normocholesterolaemic and hypercholesterolaemic rats. METHODS Triglycerides (TG), total, LDL- and HDL-cholesterol levels, total antioxidant capacity (FRAP and ABTS assays), malondialdehyde (MDA) and protein carbonyls were analysed in serum, and MDA, glutathione and antioxidant enzyme activity in livers of rats drinking water or mate fed normal or cholesterol-cholic supplemented diets. RESULTS ABTS, glutathione and antioxidant enzymes were not affected by any treatment. In normocholesterolaemic animals, mate had no effect on serum lipids or antioxidant status, yet it increased serum carbonyls and liver MDA concentrations. In hypercholesterolaemic rats, mate consumption had no effect on HDL-cholesterol or protein carbonyls, yet it showed a marked hypolipidaemic action, decreasing TG, total and LDL-cholesterol, and serum MDA levels that had been increased after consuming the high-cholesterol diet. CONCLUSION Potential beneficial effect of mate on markers of cardiovascular risk seems to be restricted to hyperlipaemic animals.

Pharmacokinetics of caffeine and its metabolites in plasma and urine after consuming a soluble green/roasted coffee blend by healthy subjects

Coffee is widely consumed worldwide; therefore, the methylxanthines contained in coffee, mainly caffeine (CF), are among the most abundant bioactive compounds in our diet. In the present work, the bioavailability and metabolism of methylxanthines in a commercial soluble green/roasted coffee blend was studied. After a 3-day restriction of methylxanthine-containing foods, fasting healthy subjects (12 men and women) consumed the coffee product containing 70.69mg CF and 0.119mg theobromine (TB). Plasma samples were taken before (t=0h) and after coffee consumption at different time points (0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, 9, 10 and 12h). Urine was collected at baseline (-2-0h) and at different intervals (0-2, 2-5, 5-8, 8-12 and 12-24h). Samples were analyzed by HPLC-DAD and LC-MS-QToF, and pharmacokinetic parameters were calculated. CF was the main methylxanthine found in plasma (Cmax=10.50μM, Tmax=1.2h). In addition, seven methylxanthines and methyluric acids were detected between 0.5 and 12h after coffee intake, paraxanthine (PX) being the major metabolite (Cmax=3.36μM), followed by 1-methyluric acid (1-MU; Cmax=1.44μM) and 1-methylxanthine (1-MX; Cmax=1.27μM), identified in plasma samples for the first time. In 24h urine, eleven methylxanthines and methyluric acids were detected, 1-MU being the major metabolite (Cmax=150.52μM, Tmax=12h) amounting to 67.7% of the total urinary metabolites. In conclusion, a rapid absorption, metabolization and excretion of caffeine and its derived methylxanthines and methyluric acids have been observed after consumption of a green/roasted coffee product.

Exhaustive Qualitative LC-DAD-MSn Analysis of Arabica Green Coffee Beans: Cinnamoyl-glycosides and Cinnamoylshikimic Acids as New Polyphenols in Green Coffee

Coffee is one of the most consumed beverages in the world, due to its unique aroma and stimulant properties. Although its health effects are controversial, moderate intake seems to be beneficial. The present work deals with the characterization and quantification of polyphenols and methylxanthines in four Arabica green coffee beans from different geographical origins. The antioxidant activity was also evaluated. Forty-three polyphenols (cinnamic acid, cinnamoyl-amide, 5 cinammoyl-glycosides, and 36 cinnamate esters) were identified using LC-MSn. Among these, cinnamate esters of six different chemical groups (including two dimethoxycinnamoylquinic acid isomers, three caffeoyl-feruloylquinic acid isomers, caffeoyl-sinapoylquinic acid, p-coumaroyl-feruloylquinic acid, two caffeoylshikimic acid isomers, and trimethoxycinnamoylshikimic acid) in addition to five isomers of cinnamoyl-glycosides called caffeoyl-2,7-anhydro-3-deoxy-2-octulopyranosic acid (CDOA) are described for the first time in Arabica green coffee beans. Moreover, 38 polyphenols (6-7% w/w) and 2 methylxanthines (1.3% w/w) were quantified by HPLC-DAD. Caffeoylquinic was the most abundant group of compounds (up to 85.5%) followed by dicaffeoylquinic and feruloylquinic acids (up to 8 and 7%, respectively) and the newly identified cinnamoyl-glycosides (CDOA) (up to 2.5%). Caffeine was the main methylxanthine (99.8%), with minimal amounts of theobromine (0.2%). African coffees (from Kenya and Ethiopia) showed higher polyphenolic content than American beans (from Brazil and Colombia), whereas methylxanthine contents varied randomly. Both phenols and methylxanthines contributed to the antioxidant capacity associated with green coffee, with a higher contribution of polyphenols. We conclude that green coffee represents an important source of polyphenols and methylxanthines, with high antioxidant capacity.

Dihydrocaffeic acid, a major microbial metabolite of chlorogenic acids, shows similar protective effect than a yerba mate phenolic extract against oxidative stress in HepG2 cells

The hepatoprotective effect of a yerba mate phenolic extract (YMPE), rich in chlorogenic acids, and its main circulating metabolites dihydrocaffeic (DHCA) and dihydroferulic (DHFA) acids were assessed in human hepatoma HepG2 cells subjected to oxidative damage induced by tert-butylhydroperoxide (t-BOOH). Direct treatment of HepG2 cells with realistic concentrations of YMPE (1, 10 and 50μg/mL), DHCA or DHFA (0.2, 1, 10μM) for 20h was not cytotoxic and significantly decreased ROS generation. Pre-treatment with YMPE and DHCA prevented the cytotoxicity and macromolecular damage induced by t-BOOH. Moreover, decreased levels of reduced glutathione (GSH), and increased ROS levels and antioxidant enzyme activity induced by t-BOOH were dose-dependently recovered. DHFA only showed a slight protection against cell cytotoxicity, lipid oxidation and GSH depletion. In conclusion, YMPE and one of its major microbial metabolites, DHCA, confer significant protection against oxidative damage, adding evidences to the beneficial health effects associated with mate intake.

An aqueous pomegranate seed extract ameliorates oxidative stress of human hepatoma HepG2 cells

BACKGROUND Aqueous pomegranate seed extract (PSE), a by-product of the pomegranate juice industry, was recently identified as a potential antiglycative ingredient. Ellagic acid was proposed as the major polyphenol responsible for the antiglycative activity as exerted in in vitro models. However, there is no information on safety aspects of this extract in biological systems before its application as ingredient. The cytotoxicity of PSE (1-100 µg mL(-1) ) was evaluated by determining its effect on cell viability and redox status of cultured HepG2 cells. The protective effect of the PSE against oxidative stress induced by tert-butyl hydroperoxide (t-BOOH) was also investigated. RESULTS No changes in cell integrity or intrinsic antioxidant status resulted from a direct treatment with aqueous PSE, even at high dosage. In addition, reactive oxygen species (ROS) induced by t-BOOH were reduced by 21% when cells were pretreated with 100 µg mL(-1) of aqueous PSE at 180 min. The range of concentrations investigated was effective in decreasing the ROS formation but not in a dose-dependent manner. CONCLUSION Aqueous pomegranate seed extract enhances human hepatoma cells integrity and resistance to cope with a stressful situation at concentration up to 100 µg mL(-1) .

Improved LC-MSn characterization of hydroxycinnamic acid derivatives and flavonols in different commercial mate (Ilex paraguariensis) brands. Quantification of polyphenols, methylxanthines, and antioxidant activity

Yerba mate is a beverage rich in bioactive compounds popular in South America. Polyphenols and methylxanthines were qualitatively and quantitatively analyzed in four commercial brands of yerba mate, as well as the antioxidant capacity of the beverages. Using LC/MSn analysis, 58 polyphenols were observed of which 4-sinapoylquinic acid, di- and tri-methoxycinnamoylquinic acids, two isomers of trimethoxycinnamoylshikimic acid and four isomers of caffeoyl-2,7-anhydro-3-deoxy-2-octulopyranosonic acid were identified for the first time in mate. Additionally, 46 polyphenols and 2 methylxanthines were quantified by HPLC-DAD. Hydroxycinnamic acid derivatives and flavonols comprised 90% and 10% of mate phenols, respectively, 3-caffeoylquinic (26.8-28.8%), 5-caffeoylquinic (21.1-22.4%), 4-caffeoylquinic (12.6-14.2%) and 3,5-dicaffeoylquinic acids (9.5-11.3%) along with rutin (7.1-7.8%) were the most abundant polyphenols, whereas caffeine was the main methylxanthine (90%). Ilex paraguariensis is an important source of polyphenols with moderate methylxanthines content; therefore its high antioxidant capacity was mainly associated to its polyphenolic composition.