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It is shown that a transition from traditional C18 (I) stationary phases with non-polar endcapping to the C10CN (II) phase, containing terminal polar groups, leads to a change in selectivity comparable to an increase in the activity of residual silanol groups in phases I. The effect was detected in the separation of isomeric monocaffeoylquinic acids. Two versions of the gradient mode are proposed using a Diasfer-110-C10CN column and water–acetonitrile components of the mobile phase acidified with H3PO4 for the separation of chlorogenic acids and caffeine in green coffee extracts from different manufacturers. It was shown that the proposed chromatographic method can also be used to determine trigonelline, the retention of which significantly increased in replacing phase I with phase II. The proposed method was used to differentiate the fruits of two types of coffee—Arabica and Robusta. It was found that the total amount of chlorogenic acids and caffeine is higher in Robusta coffee extracts. [ABSTRACT FROM AUTHOR]

Coffea spp. are remarkable sources of phytochemicals, but the lack of a well-defined culture medium aimed at the induction of non-embryogenic and friable callus hampers the establishment of plant cell suspension cultures for large-scale production of valuable compounds. In this paper, we describe a one-medium protocol suitable to obtain both callus and cell suspension cultures from leaves of two elite cultivars of C. arabica. The protocol was developed through an iterative process involving the determination of the best concentration of auxin and cytokinin, their optimal ratio, as well as the most effective molecule of either hormone class. Young leaves were found to be a good and easy-to-use explant source for callus induction and proliferation, provided that a cytokinin was present in association with a chlorinated auxin in a full strength, semi-solid MS medium. The best results were obtained by hormone concentration and combination of 1 mg/L of both kinetin and 2,4,5-trichlorophenoxyacetic acid. The same ratio of these growth regulators was conveniently used for the development and stabilization of cell suspension cultures in liquid MS medium. When grown in darkness, stabilized suspension cultures showed a fine and homogeneous texture, with a 10-fold biomass increase within 25 days and a cell viability > 90%. In addition, the phytochemical profile revealed the presence of the most widely studied coffee compounds. The protocol can be applied to obtain adequate amounts of cell biomass for use in physiological studies concerning the production of secondary metabolites.Key messages: A protocol for callus induction and undifferentiated cell suspensions establishment in Coffea arabica was developed. This technique represents a useful tool for in vitro production of remarkable phytochemicals from coffee. [ABSTRACT FROM AUTHOR]

Introduction: Coffee is a complex brew that contains several bioactive compounds and some of them can influence blood pressure (BP) and endothelial function (EF), such as caffeine and chlorogenic acids (CGAs). Aim: This study aimed to evaluate the acute effects of coffee on BP and EF in individuals with hypertension on drug treatment who were habitual coffee consumers. Methods: This randomized crossover trial assigned 16 adults with hypertension to receive three test beverages one week apart: caffeinated coffee (CC; 135 mg caffeine, 61 mg CGAs), decaffeinated coffee (DC; 5 mg caffeine, 68 mg CGAs), and water. BP was continuously evaluated from 15 min before to 90 min after test beverages by digital photoplethysmography. Reactive hyperemia index (RHI) assessed by peripheral arterial tonometry evaluated EF before and at 90 min after test beverages. At the same time points, microvascular reactivity was assessed by laser speckle contrast imaging. Repeated-measures-ANOVA evaluated the effect of time, the effect of beverage, and the interaction between time and beverage (treatment effect). Results: Although the intake of CC produced a significant increase in BP and a significant decrease in RHI, these changes were also observed after the intake of DC and were not significantly different from the modifications observed after the consumption of DC and water. Microvascular reactivity did not present significant changes after the 3 beverages. Conclusion: CC in comparison with DC and water neither promoted an acute increase in BP nor produced an improvement or deleterious effect on EF in individuals with hypertension on drug treatment who were coffee consumers. [ABSTRACT FROM AUTHOR]

The bioactive ingredients of green coffee beans were extracted using high pressure-assisted extraction (HPE) and compared against those derived from the traditional heat reflux (HR) extraction method. The effect of extraction conditions on the extraction efficiency of functional components (chlorogenic acids, caffeine, total phenols, and flavonoids); the free radical-scavenging activity of the extract; and the inhibition of α-glucosidase, α-amylase, and food pathogenic bacteria were measured. Using water as the solvent, the high-pressure extraction rates of chlorogenic acid (CGA) and caffeine were 3.20–5.75 mg/100 g and 4.32–7.95 mg/100 g, respectively, which were significantly higher than the rates obtained using the traditional method (2.92 mg/100 g and 3.54 mg/100 g, respectively). Under optimal extraction conditions of 25 °C and 600 MPa for 2.5 min, inhibition of α-glucosidase and α-amylase by the extract reached rates of 27.8% and 26.7%, respectively, with the highest free radical-scavenging activity of 81.5%. HPE exhibited antibacterial activity against both gram-positive (Staphylococcus aureus and Listeria innocua) and gram-negative (Escherichia coli and Salmonella enterica) bacteria, and the lowest minimum inhibitory concentration (4.69 mg/mL) against S. aureus was achieved. Scanning electron microscopic observations evidenced that high pressure partially damaged the intercellular compartments of coffee beans, increased solvent permeability, and improved the extraction rate of bioactive ingredients. The outcomes of the study demonstrated that HPE can be employed as an efficient extraction technique for production of chlorogenic acids and caffeine that might have a potential application in food and related industries. [ABSTRACT FROM AUTHOR]

To reveal the molecular mechanism in the accumulation of chlorogenic acids (CGAs) in peach (Prunus persica) fruit during growth and development, CGA contents in the flesh of the three peach cultivars 'Ruiguang 18', 'Heiyoutao', and 'Beijingyixianhong' were determined. The expression levels of CGA metabolism-related genes were analyzed based on transcriptome data (RNA-seq). These candidate genes were then screened and real-time fluorescent quantitative PCR (qRT-PCR) was performed to verify their expression. The results showed that the content of total CGAs, 5-O-caffeoylquinic acid and 3-O-caffeoylquinic acid, in the flesh of 'Ruiguang 18' exhibited a decreasing trend during fruit development, and there was a great drop at maturity stage (P < 0.05). The three contents in 'Heiyoutao' increased first at stage S2 (P < 0.05) and then decreased significantly (P < 0.05). In 'Beijingyixianhong', they stayed stable in the early stages, then total CGAs and 3-O-caffeoylquinic acid decreased significantly at the maturity stage (P < 0.05). RNA-seq transcriptome data analysis and qRT-PCR expression analysis showed that the accumulation of CGAs in fruit flesh was mainly affected by the expression of Prupe.3G100800 (PpHCT) and Prupe.3G107300 (Pp4CL), and their expression levels were highly consistent with total CGA content. Thus, we concluded that Prupe.3G100800 (PpHCT) and Prupe.3G107300 (Pp4CL) are the key genes for CGAs synthesis in peach flesh. [ABSTRACT FROM AUTHOR]

In this work, the electrochemical determination of antioxidant compounds in green coffee beans using a low-pressure chromatographic system based on a single 1-cm length monolithic column is presented. Screening of the antioxidant compounds in green coffee beans using this approach was performed under different elution conditions, considering different supporting electrolyte media and applied potential values at detection. Mass spectra analyses for the structure elucidation of the separated compounds are presented. Under the selected experimental conditions, five chlorogenic acids (3-, 4-, 5-, cis-5-caffeoylquinic acids, and 5-feruloylquinic acid) and caffeine were identified. Resolution values higher than 1.5 were achieved for all identified compounds. The limits of detection for 5-caffeoylquinic acid and caffeine were 9.25 µmol L−1 and 6.73 µmol L−1, respectively. Acetonitrile consumption was of 0.325 mL per analysis. The figures of merit obtained plus the simplicity of the analytical system assembling and handling highlight the high competitiveness of the presented low-cost approach to be used in substitution of HPLC methodologies currently performed to this end, and for further analytical applications in complex matrices. [ABSTRACT FROM AUTHOR]

The fruits of several species of the Aronia genus were studied in the content of anthocyanins and chlorogenic acids. It was found that errors in determining the Aronia species can be excluded by using quantitative ratios between the main anthocyanins—cyanidin-3-galactoside (Cy3Gala, the main component), cyanidin-3-arabinoside (Cy3Ara), cyanidin-3-glucoside (Cy3Glu) and cyanidin-3-xyloside (Cy3Xyl). The highest level of anthocyanin accumulation was found for Aronia melanocarpa (Michx.) Elliott fruits. Meanwhile for the species differentiation the least accumulation in fruits of Cy3Glu (no more than 0.8% of the total anthocyanins) and Cy3Xyl (not more than 1.5 mol % of the amount of anthocyanins) may explored in comparison with that of A. prunifolia (Marshall) Rehder and A. mitschurinii A.K. Skvortsov & Maitul., the latter being often mistaken for black chokeberry. Even more reliable for the differentiation is the content and ratio of isomeric chlorogenic acids in the fruits. So, fruits of A. melanocarpa accumulate as the main chlorogenic acid 3-caffeoylquinic acid (3CQA, about 69.4 mol % of the sum of chlorogenic acids) and, in smaller but comparable amounts 5-caffeoylquinic (5CQA) and 4-caffeoylquinic (4CQA) acids, which account for about 14 mol %. In fruits of other species of chokeberry accumulation levels of 3CQA and 5CQA are comparable, but may vary with the change of the dominant one, while the accumulation 4CQA is characteristically low (less than 3.5 mol %). The paper discusses and experimentally confirms some errors in the sample preparation of Aronia fruit samples before the qualitative and quantitative determination of anthocyanins, and proposes a method of excluding them. [ABSTRACT FROM AUTHOR]

LC-HRMS Q-TOF method was used to identify and quantify metabolites of Foleyola billotii Maire plant. A series of 15 compounds were tentatively identified: three glucosinolates, 2 aliphatic (progoitrin and gluconapin) and 1 indolic (glucobrassicin); six chlorogenic acids, four coumaroyl quinic acid (CoQA) isomers and two feruloyl quinic acid (FQA) isomers; two diglycosylated flavonols, one quercetin-derived and one kaempferol-derived; three sinapic acid derivatives, sinapoyl threonic acid, hexose sinapate and a third derivative supposedly attributed to a feruloyl quinic acid sinapate; and finally, a glycosylated trans-cinnamic acid which could correspond to feruloylthreonic acid (FTA). Glucosinolates content was 55 times higher in cultivated plants; chlorogenic acids and flavonols increased too by 5 and 3.5 times, respectively. In contrast, sinapates decreased by 3 times in cultivated plants, this last result can be explained by reduced UV-B intensity in the north of Algeria. The change in F. billotii compound profile was more quantitative rather than qualitative. [ABSTRACT FROM AUTHOR]

The spent coffee ground is a by-product of the coffee industry with high potential because of its beneficial properties for health, however, due to high water content, it is necessary to stabilize it without affecting bioactive capacities. The main objective of this work was to evaluate the effect of different convective drying conditions, on some technological and functional properties of the spent coffee ground and to determine the thermal degradation kinetics of chlorogenic acids. The methodology included the optimization of the drying process conditions of spent coffee ground: temperature (40–60 °C), air flow (1.0–2.0 m/s) and cake thickness (0.01–0.02 m). Effective diffusion coefficient, moisture, water activity, water and oil holding capacity, antioxidant capacity, caffeine, and seven chlorogenic acids were determined. The results showed that the best drying conditions were 60 °C, 2.0 m/s, 1.28 cm which allows retaining 84.24% of total polyphenols and 66.00% of the antioxidant capacity. Chlorogenic acids showed thermal degradation kinetics of first order under the optimal process conditions. In general, it is concluded that the convective drying process is a valid technique for processing of coffee grounds, as it allows the preservation of antioxidant compounds with a potentially beneficial effect on health, providing a stabilized low moisture content that can be used in food applications. [ABSTRACT FROM AUTHOR]

Coffee infusion experiments were conducted to infer how operational variables (time, temperature, mass to volume ratio, and grinding) might affect the efficiency and/or selectivity of compounds extraction. Although the different variables have extensively been reported independently, to the best of our knowledge, no experimental design was yet delineated to study the simultaneous effect of variables in coffee composition. This study fulfills this gap by constructing surface models that reflect the responses in a wide-ranging design space. The freeze-dried extracts were compared regarding the overall yield of extraction, carbohydrate content and composition, caffeine, chlorogenic acid (5-CQA) content, color, and viscosity. Temperature was the major factor for coffee extracts differentiation, regarding both overall and carbohydrates yield and composition. The extraction process efficiency is more related to galactomannans extraction than arabinogalactans. Varying operational conditions, coffee extracts with distinct chemical properties are obtained from the same roasted coffee, broadening their applications in food formulations. [ABSTRACT FROM AUTHOR]

The selectivity of the separation of isomeric chlorogenic acids often found in natural samples (3, 4, and 5-caffeoylquinic acids, 3CQA, 4CQA, and 5CQA, respectively) under the conditions of reversed-phase chromatography on stationary phases of different brands, Kromasil (C18, C8, and C4), Symmetry C18, Reprosil-Pur C18-AQ, Nucleosil RPC18, and Diasfer C18, is studied. The results obtained favor the retention mechanism with predominant surface adsorption. The change in the selectivity of separation of a pair of 5CQA and 4CQA correlates with a change in the activity of residual silanol groups. At their low activity, the order of elution 3CQA < 5CQA < 4CQA does not depend on the mode (isocratic or gradient), the type of organic modifier, and the addition of an acid (formic, orthophosphoric) to the mobile phase. With an increase in the activity of silanol groups, the separation of the pair of 5CQA and 4CQA deteriorates up to the inversion of retention times. A Kromasil 100-5C18 column (250 × 4.6 mm) was chosen for a reliable separation of three acids in plant samples (instant coffee, juice, and "Mate" tea). [ABSTRACT FROM AUTHOR]

Purpose: The purpose of this study was to evaluate acute effects of coffee with a high content of chlorogenic acids and different hydroxyhydroquinone contents on postprandial endothelial dysfunction. Methods: This was a single-blind, randomized, placebo-controlled, crossover-within-subject clinical trial. A total of 37 patients with borderline or stage 1 hypertension were randomized to two study groups. The participants consumed a test meal with a single intake of the test coffee. Subjects in the Study 1 group were randomized to single intake of coffee with a high content of chlorogenic acids and low content of hydroxyhydroquinone or coffee with a high content of chlorogenic acids and a high content of hydroxyhydroquinone with crossover. Subjects in the Study 2 group were randomized to single intake of coffee with a high content of chlorogenic acids and low content of hydroxyhydroquinone or placebo coffee with crossover. Endothelial function assessed by flow-mediated vasodilation and plasma concentration of 8-isoprostanes were measured at baseline and at 1 and 2 h after coffee intake. Results: Compared with baseline values, single intake of coffee with a high content of chlorogenic acids and low content of hydroxyhydroquinone, but not coffee with a high content of chlorogenic acids and high content of hydroxyhydroquinone or placebo coffee, significantly improved postprandial flow-mediated vasodilation and decreased circulating 8-isoprostane levels. Conclusions: These findings suggest that a single intake of coffee with a high content of chlorogenic acids and low content of hydroxyhydroquinone is effective for improving postprandial endothelial dysfunction. Clinical Trial Registration: URL for Clinical Trial: https://upload.umin.ac.jp; Registration Number for Clinical Trial: UMIN000013283. [ABSTRACT FROM AUTHOR]

Abstract: Chlorogenic acid and its derivatives (CADs) are the bioactive components in Gardenia jasminoides Ellis (GJE) fruits but occur at a low level. In the current paper, four types of calluses were induced from GJE leaf explants, and light yellow-green calluses with a fine appearance and texture were selected for the development of a cell suspension culture to produce CADs. Additionally, cultured cells were elicited by salicylic acid (SA) and methyl jasmonate (MeJA) to increase the CADs yield. Seven CADs compounds were identified via HPLC-TOF-MS/MS, namely, 3-caffeoylquinic acid, 4-caffeoylquinic acid, 5-caffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, 3,5-dicaffeoyl-4-O-(3-hydroxyl-3-methyl)-glutaroylquinic acid, and malonyl-4,5-O-dicaffeoylquinic acid. This is the first identification of malonyl-4,5-O-dicaffeoylquinic acid from GJE. Both SA and MeJA significantly promoted the CADs yield, but MeJA was more effective than SA. Cells elicited by 200 µM MeJA showed the maximum total CADs content and yield, up to 20.98 ± 1.1 mg g−1 and 232.32 ± 9.1 mg L−1, which were 19.51 times and 14.14 times those of the control, respectively. Furthermore, the total CADs extracted from cells elicited by 200 µM MeJA displayed a much higher antioxidant capacity than that of natural GJE fruits. Our research suggested that a GJE cell culture combined with MeJA elicitation has a promising application in natural CADs production.Graphical Abstract: [ABSTRACT FROM AUTHOR]

Purpose: Yerba maté is widely consumed in South America as different beverages, such as maté tea (roasted leaves) and chimarrão (green dried leaves), and linked to health benefits, mainly attributed to chlorogenic acids (CGAs). Health effects of CGAs depend on their bioavailability, but such data are scarce. The aim of this study was to investigate the distribution of CGAs and metabolites in tissues, hepatic and plasmatic kinetic profile and urinary excretion after ingestion of maté tea or 5-caffeoylquinic acid (5-CQA). Methods: Wistar rats ingested maté tea (MT) or 5-CQA (ST) and were killed after 1.5 h for tissue distribution analysis (pilot study) or at 0.5, 1, 2, 4 and 8 h for liver and plasma kinetics (main experiment). Urine was collected in metabolic cages. Biological samples were analyzed by UPLC-DAD-MS with and without incubation with β-glucuronidase and sulfatase. Results: CGAs and metabolites were detected in all tissues. Caffeic acid was the main compound in plasma up to 2 h after ingestion of maté tea, while 5-CQA predominated in ST group. Concentration of microbial metabolites increased 4 h after gavage and reached higher amounts in MT plasma and liver, when compared to ST group. Approximately 4.0 % of compounds ingested by MT and 3.3 % by ST were recovered in urine up to 8 h after the gavage. Conclusion: The study confirms that not only absorption, but also metabolization of CGAs begins in stomach. There were differences in compounds formed from maté tea or isolated 5-CQA, showing that CGAs profile in food may influence qualitatively and quantitatively the metabolites formed in the body. [ABSTRACT FROM AUTHOR]

In this study, flow-through chronopotentiometry (FTCP) has been developed as an electroanalytical method for characterization (identification and quantification) of chlorogenic acids (CGAs) in coffees. The characterization of CGAs in coffee was based on the electrochemical behavior of the main chlorogenic acid (CGAs) isomers presented in coffee (caffeoylquinic acids (CQAs), dicaffeoylquinic acids (diCQAs), and feruloylquinic acids (FQAs)) and the spiking of CGAs standards in coffee samples. The FTCP study has shown that electrochemical properties of CGAs strongly depend on their chemical structure and electronic properties, particularly on the presence of electron-donating −OH, −CH═CH− and −OCH groups and strong electron-withdrawing ester (−COOR) group presented in their structure. The FTCP measurements of coffee samples show that their electrochemical behavior is very similar to that of CGAs. Therefore, FTCP can be used for characterization of CGAs and determination of their content in coffees. 5- O-Caffeoylquinic acid (5-CQA), prevailed CGAs in coffees, was used as a standard for quantification of total CGA content in coffee. The linear calibration curve of 5-CQA was observed within the concentration range of 5 to 100 μmol L with the limit of detection 5.7·10 mol L. The total CGA content of coffees has been expressed in 5-CGA equivalents per 100 g of coffee. It was shown that FTCP is a very sensitive, precise, and acurate method for determination of total CGA content in coffee. It should be noted that in presented investigation, FTCP was for the first time used for the study of electrochemical properties of polyphenolic antioxidants (including CGAs) and their characterization in some of the food samples. [ABSTRACT FROM AUTHOR]

Brazil is the largest producer and exporter of coffee in the world; the beverage is recognized as the main source of chlorogenic acids (CA), which have beneficial effects on human health. In this study, a multivariate optimization of extraction conditions was performed, aiming to maximize the extraction of chlorogenic acids isomers (3-caffeoylquinic acid (3-CQA), 4-caffeoylquinic acid (4-CQA), 5-caffeoylquinic acid (5-CQA), 3.4-dicaffeoylquinic acid (3.4-DQA), 3.5-dicaffeoylquinic acid (3.5-DQA), and 4.5-dicaffeoylquinic acid (4.5-DQA)). The optimal extraction condition was obtained using ethanol and water in the proportions of 32.4:67.6, and a 22-min extraction period, resulting in a fast, precise, accurate, and cost-effective method, with low toxic waste generation. Optimized extraction conditions were employed to investigate the CA content in different types of ground roasted coffee available on the market (traditional, extra strong, gourmet, decaffeinated, and organic). Results showed that, although these products are differentiated by their value of trade, no difference in the chlorogenic acids content was found. [ABSTRACT FROM AUTHOR]

Chlorogenic acids from coffee show high antioxidant activity that contributes to inhibition of oxidative stress, which results in anticancer, antidiabetic and antiobesity coffee activities. Coffee roasting causes their partial decomposition and partial incorporation into Maillard reaction products also of high antioxidant activity, and it is still not clear whether more favorable impact on health is associated with the consumption of green or roasted coffee. The aim of the study was to evaluate the impact of coffee roasting process and also of the degree of chlorogenic acid esterification on bioactivity of coffee extracts. Extracts of green and roasted coffee and their fractions containing mono- and dichlorogenic acids (six extracts) were obtained by purification and optional fractionation of crude extracts using preparative centrifugal partition chromatography technique. The extracts were evaluated in vitro in terms of scavenging capacity, influence on lipids accumulation in 3T3L1 adipocytes, cytoprotective potential decreasing oxidative stress induced by cellular pro-oxidants in βTC3 cells, and in vivo by selected physiological indices in rats fed diets supplemented with the extracts. Roasted coffee extracts, in particular hydrophilic fraction containing monochlorogenic acids and Maillard reaction products, showed the highest scavenging capacity and contributed to the highest level of antioxidants in the hydrophilic fraction of rats serum and may be potentially used as anticancer nutraceutical with oxidative stress inhibitor activity. Non-fractionated green coffee extract exhibited antidiabetic activities including inhibition of adipogenesis, while in vivo it significantly lowered the levels of triglycerides, glucose and oxidized glutathione. [ABSTRACT FROM AUTHOR]

Main Conclusion: This work shows that, in tobacco, the ectopic expression of VvMYBPA1 , a grape regulator of proanthocyanidin biosynthesis, up- or down-regulates different branches of the phenylproanoid pathway, in a structure-specific fashion. Proanthocyanidins are flavonoids of paramount importance for animal and human diet. Research interest increasingly tilts towards generating crops enriched with these health-promoting compounds. Flavonoids synthesis is regulated by the MBW transcriptional complex, made of R2R3MYB, bHLH and WD40 proteins, with the MYB components liable for channeling the complex towards specific branches of the pathway. Hence, using tobacco as a model, here, we tested if the ectopic expression of the proanthocyanidin regulator VvMYBPA1 from grape induces the biosynthesis of these compounds in not-naturally committed cells. Here, we show, via targeted transcriptomic and metabolic analyses of primary transgenic lines and their progeny, that VvMYBPA1 alters the phenylpropanoid pathway in tobacco floral organs, in a structure-specific fashion. We also report that a modest VvMYBPA1 expression is sufficient to induce the expression of both proanthocyanidin-specific and early genes of the phenylpropanoid pathway. Consequently, proanthocyanidins and chlorogenic acids are induced or de novo synthetised in floral limbs, tubes and stamens. Other phenylpropanoid branches are conversely induced or depleted according to the floral structure. Our study documents a novel and distinct function of VvMYBPA1 with respect to other MYBs regulating proanthocyanidins. Present findings may have major implications in designing strategies for enriching crops with health-promoting compounds. [ABSTRACT FROM AUTHOR]

The beverage obtained from the yerba mate tea, besides being the most consumed in Brazil, has high concentrations of chlorogenic acids. In this study, a central composite design was employed to establish the best infusion time, temperature and water volume to maximize the extraction of chlorogenic acids 5-caffeoylquinic (5CQ), 3.4-dicaffeoylquinic (3.4 DQ), 3.5-dicaffeoylquinic (3.5 DQ) and 4.5-dicaffeoylquinic (4.5 DQ), from the leaves and stems of yerba mate tea (beverage ready for consumption). Analyses were performed by high-performance liquid chromatography and the optimum conditions were obtained through the use of the desirability function of Derringer and Suich. The maximum chlorogenic acids content in the beverage was obtained when the infusion was prepared with 2 g of mate tea, in 300 mL of water at 95 °C, under infusion for 16 min. The optimal conditions were applied for the preparation of beverages from 15 commercial samples of yerba mate tea, and it was observed that the sum of the concentration of the four compounds showed variation of up to 79 times between the average of the samples, which can be attributed to climatic conditions of cultivation of the plant and/or of processing. [ABSTRACT FROM AUTHOR]

Green coffee is one of health-promoting supplements of the diet, applied in the form of either preparations or enriched food products. Its positive impact is manifested by mitigation of the development of certain tumors, e.g., in the colon and liver, and type 2 diabetes. Many studies proved that chlorogenic acids are the main active substances in green coffee. The bioavailability of these compounds depends among others on their interactions with other components of the diet, mainly proteins. When they are used as food ingredients, their bioavailability is additionally decreased because of the decomposition or interactions with other ingredients during food processing. The undesirable changes may be limited among others by microencapsulation, for example with β-cyclodextrin. In this study, rats were fed the pro-oxidative high fat diet, which was supplemented with chlorogenic acids from green coffee that were used in four forms such as: a purified extract, complexes of chlorogenic acids and β-cyclodextrin, and bread supplemented with either the extract or the β-cyclodextrin inclusion complex. Chlorogenic acids added to bread because of the reduced absorption from the crumb in the small intestine and increased passage to the colon, contributed to the beneficial modification of enzymatic activities of intestinal microbiota. When added directly to the diet, they contributed to the improved antioxidant status in the liver and kidneys, lowered glucose level and increased HDL level. A high ratio of reduced to oxidized glutathione in the liver and a high concentration of antioxidants in the blood serum were observed after administration of chlorogenic acids in the form of inclusion complexes with β-cyclodextrin, indicating that microencapsulation increased their bioaccessibility due to the limited interactions with other components of the diet. [ABSTRACT FROM AUTHOR]

Coffee is the main source of chlorogenic acids (CGAs) in human nutrition. In this study, one main point of interest was whether the simultaneous consumption of coffee and food affects the absorption and bioavailability of CGAs. Fourteen healthy participants consumed pure instant coffee (T1), or coffee with either a high-carbohydrate (T2) or a high-fat meal (T3). All volunteers consumed the same quantity of CGAs (3.1 mg CGA/kg bw), and blood and urine samples were collected at various time points for up to 15 h and 24 h after consumption, respectively. Existing CGAs, and their respective metabolite concentrations, were determined using HPLC-ESI-MS/MS. The area under the curve (AUC) was the measure of CGA quantity in plasma samples collected after each treatment. We observed significantly greater CGA bioavailability after pure instant coffee consumption (T1) than with an additional consumption of a high-fat meal (T3). However, the difference in CGA bioavailability between pure coffee (T1) and coffee plus high-carbohydrate meal (T2) was not statistically significant. The latter observation held true when T3-samples were compared with T2-samples. When the metabolites were split into their respective classes, significant differences in the sums of AUC were observed only for some classes, and not among the treatments. Data did not show differences in the total bioavailability, but revealed differences in the kinetic of release. The co-ingestion of breakfast favored a slow and continuous release of colonic metabolites in contrast to the non-metabolized coffee components appearing in the first hour after coffee consumption. Urine samples collected over 24 h did not show any statistically significant differences among the treatments. Only samples collected within the first 6 h post-coffee consumption showed that the CGA quantities in urine samples collected after T1 and T3 were significantly greater than quantities with an additional consumption of carbohydrates (T2). Breakfast had no significant effect on CGA absorption from coffee. However, a shift in gastrointestinal transit time and in the plasma metabolite composition was observed. So breakfast consumption induces retarded release of chlorogenic acid metabolites in humans. [ABSTRACT FROM AUTHOR]

Purpose: Epidemiological evidence suggests that coffee consumption is associated with a lower risk of type 2 diabetes. Coffee contains caffeine and several other components that may modulate glucose regulation. The chlorogenic acids (CGA) in coffee have been indicated as constituents that may help to normalize the acute glucose response after a carbohydrate challenge. The aim of this study was to investigate whether two coffee beverages that differ in CGA content due to different roasting degrees will differentially affect glucose regulation. Methods: In a controlled crossover trial, 11 healthy fasted volunteers consumed 300 mL of either light (LIR) or dark (DAR) roasted coffee, or water, followed 30 min later by a 75-g oral glucose tolerance test (OGTT). Blood samples were drawn at baseline, 30, 60, and 120 min. Differences in glucose and insulin responses and insulin sensitivity index (ISI) were analyzed. The CGA and caffeine contents in the coffees were analyzed using UPLC-MS/MS. Results: No differences in glucose area under the curve (AUC) were found between treatments. Glucose concentrations were higher at 60 min after ingestion of DAR compared with water, while ingestion of LIR showed similar glucose concentrations as ingestion of water. Insulin AUC was higher after ingestion of DAR compared with water, and both coffees raised insulin concentrations and reduced ISI compared with water, with no difference between the two coffees. Conclusion: Two coffees with different CGA contents did not differentially affect glucose or insulin responses during an OGTT, but both increased the insulin response compared with water. [ABSTRACT FROM AUTHOR]

Chlorogenic acids and caffeine are important for flavor formation as well as the health effect of green coffee brews and its extracts. The content of these compounds was determined by HPLC-DAD analysis in twelve samples of coffee from Robusta and Arabica types of different geographical origin including steamed and decaffeinated coffees. Generally, Robusta coffee extracts contain twice as much caffeine as Arabica, and its content varies from 3.41 % per dry mass in Arabica type from Laos or Rwanda to 8.16 % in Robusta coffee from Indonesia. The highest concentration of 5- O-caffeoylquinic acid (5-CQA) was obtained for both coffees from Uganda. Decaffeination process does not affect the concentration of this main chlorogenic acid, but steaming of the coffee beans with hot water produced a significant decrease in the level of 5-CQA. Antioxidant activity of coffee extracts was measured by CUPRAC and F-C assays, which really measure the reducing power of the sample components. Extracts of green coffee beans from Vietnam possessed the highest antioxidant activity in both assays. [ABSTRACT FROM AUTHOR]

Exogenous application of synthetic and natural elicitors of plant defence has been shown to result in mass production of secondary metabolites with nutraceuticals properties in cultured cells. In particular, salicylic acid (SA) treatment has been reported to induce the production of phenylpropanoids, including cinnamic acid derivatives bound to quinic acid (chlorogenic acids). Centella asiatica is an important medicinal plant with several therapeutic properties owing to its wide spectrum of secondary metabolites. We investigated the effect of SA on C. asiatica cells by monitoring perturbation of chlorogenic acids in particular. Different concentrations of SA were used to treat C. asiatica cells, and extracts from both treated and untreated cells were analysed using an optimised UHPLC-QTOF-MS/MS method. Semi-targeted multivariate data analyses with the aid of principal component analysis (PCA) and orthogonal projection to latent structures-discriminant analysis (OPLS-DA) revealed a concentration-dependent metabolic response. Surprisingly, a range of chlorogenic acid derivatives were found to be downregulated as a consequence of SA treatment. Moreover, irbic acid (3,5- O-dicaffeoyl-4- O-malonilquinic acid) was found to be a dominant CGA in C. asiatica cells, although the SA treatment also had a negative effect on its concentration. Overall SA treatment was found to be an ineffective elicitor of CGA production in cultured C. asiatica cells. [ABSTRACT FROM AUTHOR]

Coffee, the one of the most popular beverages in the world, contains many bioactive compounds especially caffeine-the natural stimulant and chlorogenic acids with antioxidative properties. Other chemicals such as diterpenes may influence human serum lipids and protect from the risk of some types of cancer. All these compounds are widely determined in Coffea arabica and robusta green and roasted coffee beans, coffee brews and instant coffees to increase food quality standards. The most important analytical methods are reviewed, including these using high-performance liquid chromatography, ultraviolet spectrophotometry and voltammetry techniques, that have been applied to quantify the major bioactive compounds of coffee samples: phenols-inter alia chlorogenic acids and derivatives; methylxanthines-caffeine; trigonelline; nicotinic acid; diterpenes; and short-chain carboxylic acids. Usage of hyphenated techniques such as liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry for identification and determination of coffee constituents is also presented. [ABSTRACT FROM AUTHOR]

Previous studies suggest that only trans-isomers of chlorogenic acid (CGA) are naturally produced. Cis-isomers have been noted in some plant tissues exposed to different mechanical processes as well as untreated tobacco leaves exposed to sunlight. Very little, however, is known about the biological significance and origin of cis-isomers. Here we show for the first time the accumulation of cis-5-caffeoylquinic acid in cultured tobacco cells treated with different inducers of plant defence (lipopolysaccharides, flagellin peptide-22, chitosan, acibenzolar- S-methyl and isonitrosoacetophenone), without exposure to UV light and with a 2-fold (on average) increase in the concentration of the pool in comparison to non-stimulated cells. Our UHPLC-Q-TOF-MS and multivariate statistical results suggest the presence of a possible biological pathway responsible for the production of cis-CGAs in tobacco plants. [ABSTRACT FROM AUTHOR]

Habitual consumption of medium amounts of coffee over the whole life-span is hypothesized to reduce the risk to develop diabetes type 2 (DM2) and Alzheimer's disease (AD). To identify putative bioactive coffee-derived metabolites, first, pooled urine from coffee drinkers and non-coffee drinkers were screened by UPLC-HDMS. After statistical data analysis, trigonelline, dimethylxanthines and monomethylxanthines, and ferulic acid conjugates were identified as the major metabolites found after coffee consumption. For quantitative analysis of these markers in body fluids, targeted methods based on stable-isotope dilution and UPLC-MS/MS were developed and applied to plasma samples from a coffee intervention study ( n = 13 volunteers) who consumed a single cup of caffeinated coffee brew after a 10-day washout period. Chlorogenic acid-derived metabolites were found to be separated into two groups showing different pharmacokinetic properties. The first group comprised, e.g., ferulic acid and feruloyl sulfate and showed early appearance in the plasma (∼1 h). The second group contained particularly chlorogenic acid metabolites formed by the intestinal microflora, appearing late and persisting in the plasma (>6 h). Trigonelline appeared early but persisted with calculated half-life times ∼5 h. The plasma levels of caffeine metabolites significantly and progressively increased 2-4 h after coffee consumption and did not reach c within the time frame of the study. The pharmacokinetic profiles suggest that particularly trigonelline, caffeine, its metabolites, as well as late appearing dihydroferulic acid, feruloylglycine and dihydroferulic acid sulfate formed from chlorogenic acid by the intestinal microflora accumulate in the plasma due to their long half-life times during habitual consumption of several cups of coffee distributed over the day. Since some of these metabolites have been reported to show antioxidant effects in vivo, antioxidant-response-element activating potential, and neuroprotective properties, respectively, some of these key metabolites might account for the inflammation- and DM2/AD risk reducing effects reported for habitual life time consumption of coffee. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Nematotoxic effect of an aqueous extract of Artemisia annua and its components caffeic acid, chlorogenic acid (5-caffeoylquinic acid, 5-CQA), artemisinin and the related semi-synthetic artesunate, was investigated on the root-knot nematode Meloidogyne incognita and the potato cyst nematode Globodera rostochiensis and on the virus-vector dagger nematode Xiphinema index. Juveniles of M. incognita and G. rostochiensis and females of X. index were exposed to 500, 250 and 125 μl ml solutions of the A. annua aqueous extract, caffeic acid, chlorogenic acid and artesunate and to 50 μl ml solution of artemisinin for 2, 4, 8 and 24 h. Egg masses of M. incognita and cysts of G. rostochiensis were exposed for 24, 48, 96 h and 1 or 2 weeks only to the extract solutions. Aqueous extract was highly effective on G. rostochiensis juveniles, whereas M. incognita juveniles were affected only at long exposure times. Adversely, egg hatch inhibition was strong on M. incognita and poor or minimal on G. rostochiensis. Females of X. index were sensitive only to long exposures to the highest extract concentration. Both caffeic and chlorogenic acid did not affect juveniles of M. incognita but were highly active on G. rostochiensis juveniles and X. index females even at the lowest concentration. Artesunate toxicity was almost zero on M. incognita and low on X. index females, but high on G. rostochiensis juveniles. Artemisinin solution was lethal to more than 50 % of G. rostochiensis juveniles within 24 h, but did not affect M. incognita juveniles and X. index females. Results suggest different roles of the tested compounds in the biocidal activity on each target nematode species. The extract of A. annua and its main phytochemicals seem to have a potential to be developed into new nematicidal formulates, though their activity should be validated in the soil. [ABSTRACT FROM AUTHOR]

The objective of the paper was to identify the number of major loci explaining caffeine content in coffee seeds. Investigations were based on previously published results: (1) Caffeine binds to chlorogenic acids in a 1:1 molecular ratio; (2) Between species, the caffeine content is correlated to the chlorogenic acid content; (3) Only a part of chlorogenic acids is bound to caffeine. Especially, the content ratio between caffeine and chlorogenic acids varied between species. For identifying the number of major loci, a quantitative trait locus (QTL) approach was carried out using an interspecific cross between two highly differentiated species- Coffea liberica dewevrei and Coffea pseudozanguebariae, the latter being a caffeine-free species. As main finding, two QTLs, i.e., RCQ1 and CQA1, were identified allowing us to explain up to 97 % of the caffeine content variance. RCQ1 explained variation of the caffeine/chlorogenic acid ratio and was genetically independent of the second QTL. The latter explained the part of the caffeine content which was dependent on the chlorogenic acid content. The findings also confirmed that only a part of chlorogenic acids were trapped by caffeine, as in wild species. [ABSTRACT FROM AUTHOR]

The preparation of a cup of coffee may vary between countries, cultures and individuals. Here, an analysis of nine different extraction methods is presented regarding analytical and sensory aspects for four espressi and five lunghi. This comprised espresso and lungo from a semi-automatic coffee machine, espresso and lungo from a fully automatic coffee machine, espresso from a single-serve capsule system, mocha made with a percolator, lungo prepared with French Press extraction, filter coffee and lungo extracted with a Bayreuth coffee machine. Analytical measurements included headspace analysis with HS SPME GC/MS, acidity (pH), titratable acidity, content of fatty acids, total solids, refractive indices (expressed in °Brix), caffeine and chlorogenic acids content with HPLC. Sensory analysis included visual, aroma, flavor and textural attributes as well as aftersensation. The technical differences in the extraction methods led to a higher concentration of the respective quantities in the espressi than in the lunghi. Regarding the contents per cup of coffee, the lunghi generally had a higher content than the espressi. The extraction efficiency of the respective compounds was mainly driven by their solubility in water. A higher amount of water, as in the extraction of a lungo, generally led to higher extraction efficiency. Comparing analytical data with sensory profiles, the following positive correlations were found total solids ↔ texture/body, headspace intensity ↔ aroma intensity, concentrations of caffeine/chlorogenic acids ↔ bitterness and astringency. [ABSTRACT FROM AUTHOR]

Phenylalanine ammonia lyase (PAL) is the first entry enzyme of the phenylpropanoid pathway producing phenolics, widespread constituents of plant foods and beverages, including chlorogenic acids, polyphenols found at remarkably high levels in the coffee bean and long recognized as powerful antioxidants. To date, whereas PAL is generally encoded by a small gene family, only one gene has been characterized in Coffea canephora ( CcPAL1), an economically important species of cultivated coffee. In this study, a molecular- and bioinformatic-based search for CcPAL1 paralogues resulted successfully in identifying two additional genes, CcPAL2 and CcPAL3, presenting similar genomic structures and encoding proteins with close sequences. Genetic mapping helped position each gene in three different coffee linkage groups, CcPAL2 in particular, located in a coffee genome linkage group (F) which is syntenic to a region of Tomato Chromosome 9 containing a PAL gene. These results, combined with a phylogenetic study, strongly suggest that CcPAL2 may be the ancestral gene of C. canephora. A quantitative gene expression analysis was also conducted in coffee tissues, showing that all genes are transcriptionally active, but they present distinct expression levels and patterns. We discovered that CcPAL2 transcripts appeared predominantly in flower, fruit pericarp and vegetative/lignifying tissues like roots and branches, whereas CcPAL1 and CcPAL3 were highly expressed in immature fruit. This is the first comprehensive study dedicated to PAL gene family characterization in coffee, allowing us to advance functional studies which are indispensable to learning to decipher what role this family plays in channeling the metabolism of coffee phenylpropanoids. [ABSTRACT FROM AUTHOR]

queous and acetone/water extracts from Hamamelis virginiana leaves were investigated to obtain a thorough insight into their phenolic composition. To secure compound integrity, a gentle extraction method including the exclusion of light was used. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analyses yielded a fingerprint including 27 phenolic constituents. Quantification of the key compounds on an equivalent basis by high-performance liquid chromatography diode-array detection (HPLC-DAD) showed that gallotannins consisting of six to 11 galloyl units constitute the main fraction, whereas procyanidins and catechin represented only a minor part. Closer inspection revealed that both extracts possess virtually the same galloyl hexose distribution, and the octagalloyl hexose represents the major tannin constituent. Additionally, eight flavonol glycosides and their corresponding aglycones quercetin and kaempferol, as well as three chlorogenic acid isomers and other hydroxycinnamic acids, were identified. Moreover, stability studies on the aqueous extract (5 °C, dark; room temperature, dark; room temperature, light) revealed that the phenolic profile underwent changes when exposed to light. Especially the gallotannins proved to be considerably unstable which may result in phytochemically altered Hamamelis leaf extracts upon transport and storage. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

The oxidation of 5- O-caffeoylquinic acid (5-CQA) was investigated in the temperature range from 161 to 188 °C. The kinetic experiments were carried out in small batch reactors made of glass, where a very thin (1.1 μm) fixed bed of powder 5-CQA was in contact with air at approximately 91.4 kPa. Aqueous 5-CQA solutions were prepared with the solid found in the reactors at different reaction times and used to measure the decrease of 5-CQA concentrations with a spectrophotometer operated at 323 nm. A shrinking core model was applied to describe the non-catalytic heterogeneous reaction between 5-CQA and oxygen. It basically assumes mass transfer of O from bulk air to the outer surface of nonporous particles of solid 5-CQA, where an irreversible reaction with no formation of solid products takes place. The proposed model correctly reproduced the obtained kinetic results in all the investigated operating conditions when an exponential reduction of external area of solid with reaction time was taken into account. Arrhenius-type expressions successfully described the dependence of the calculated global reaction and shrinking rate constants on temperature. An activation energy close to 77046 J mol was found for the slow combustion reaction of 5-CQA. [ABSTRACT FROM AUTHOR]

To identify chemical parameters that might be used as discriminators, pH, soluble solids, caffeine, trigonelline, total caffeoylquinic acids, 3- O-caffeoylquinic acid, 4- O-caffeoylquinic acid, 5- O-caffeoylquinic acid, total dicaffeoylquinic acids, 3,4- O-dicaffeoylquinic acid, 3,5- O-dicaffeoylquinic acid, 4,5- O-dicaffeoylquinic acid, total feruloylquinic acids, 3- O-feruloylquinic acid, and 5- O-feruloylquinic acid were measured in Arabica and Robusta coffees submitted to three roasting levels. It was found that the fraction of soluble solids increased with roasting level, being slightly higher in Robusta roasted coffee. The contents of caffeine did not vary significantly between roasting degrees within the Arabica and Robusta samples, respectively, revealing a considerable stability during browning. The contents of trigonelline in Arabica and Robusta coffee decreased significantly with browning intensification. Overall, the levels of chlorogenic acids remained higher in Robusta roasted coffee beans but decreased sharply with roast increase. With roasting intensification, the ratio of total caffeoylquinic acids, total dicaffeoylquinic acids, and total feruloylquinic acids varied markedly in both species, with the proportion of total caffeoylquinic acids and total feruloylquinic acids increasing significantly, whereas the opposite occurred with dicaffeoylquinic acids. One can conclude, through the application of a multivariate analysis, that these chemicals form four clusters, constituting caffeine, trigonelline, total dicaffeoylquinic acids, and total feruloylquinic acids a relevant group for T roasting level discrimination, in both coffee species. Additionally, detailing discriminators for roasting intensity in Arabica coffee might be caffeine, trigonelline, 3- O-caffeoylquinic acid, and 4- O-caffeoylquinic acid, whereas in Robusta roasted coffee are trigonelline, 3- O-caffeoylquinic acid, 4- O-caffeoylquinic acid, 5- O-caffeoylquinic acid, 3,4- O-dicaffeoylquinic acid, 3,5- O-dicaffeoylquinic acid, 4,5- O-dicaffeoylquinic acid, 3- O-feruloylquinic acid, and 5- O-feruloylquinic acid. [ABSTRACT FROM AUTHOR]

Water and ethanolic extracts were obtained from green and roasted (3 different roast degrees) Arabica and Robusta coffee beans. Three types of water extracts were prepared from the examined, finely ground material through: (a) brewing with boiling water, (b) boiling in water, and (c) boiling in water under elevated pressure. All these extracts were lyophilized. Two types of ethanolic extracts were derived from the examined material through (a) extraction of the finely ground coffee beans and (b) extraction of the solid residue that remained after boiling the coffee beans in water under elevated pressure. These ethanolic extracts were dried. Both water and ethanolic extracts were analyzed for concentration of potential antioxidants such as chlorogenic acids and caffeine (by HPLC) and Maillard reaction products (measurements of absorbance at 420 nm). Concentration of chlorogenic acids in Robusta extracts varied between 0.4 and 36.0 g × 100 g−1 dry extract weight (db.), while in Arabica extracts it ranged from 0.1 to 22.4 g × 100 g−1 db. Extracts of dark roasted Arabica contained more chlorogenic acids than those of Robusta. Concentration of caffeine, which in green and roasted coffee beans is maintained at the similar level, tended to increase in Robusta extracts with the roast degree and temperature of extraction with water, while in case of Arabica extracts there was no noticeable tendency. Caffeine concentrations varied between 0.12 and 8.41 g × 100 g−1 db. and between 0.03 and 6.53 g × 100 g−1 db. in Robusta and Arabica extracts, respectively. Ethanolic extracts were characterized by relatively higher caffeine concentrations and lower contents of brown pigments and chlorogenic acids as compared to water extracts. The richest in antioxidants were extracts of green Robusta coffee beans derived through boiling in water under elevated pressure. [ABSTRACT FROM AUTHOR]

Chlorogenic acids (CGA) are involved in the bitterness of coffee due to their decomposition in phenolic compounds during roasting. CGA mainly include caffeoyl-quinic acids (CQA), dicaffeoyl-quinic acids (diCQA) and feruloyl-quinic acids (FQA), while CQA and diCQA constitute CGA sensu stricto (CGAs.s.). In the two cultivated species Coffea canephora and Coffea arabica, CGAs.s. represents 88% and 95% of total CGA, respectively. Among all enzymes involved in CGA biosynthesis, caffeoyl-coenzyme A 3-O-methyltransferase (CCoAOMT) is not directly involved in the CGAs.s. pathway, but rather in an upstream branch leading to FQA through feruloyl-CoA. We describe how a partial cDNA corresponding to a CCoAOMT encoding gene was obtained and sequenced. Specific primers were designed and used for studying polymorphism and locating the corresponding gene on a genetic map obtained from an interspecific backcross between Coffea liberica var. Dewevrei and Coffea pseudozanguebariae. Offspring of this backcross were also evaluated for the chlorogenic acid content in their green beans. A 10% decrease was observed in backcross progenies that possess one C. pseudozanguebariae allele of the CCoAOMT gene. This suggests that CGAs.s. accumulation is dependent on the CCoAMT allele present and consequently on the activity of the encoded isoform, whereby CGA accumulation increases as the isoform activity decreases. Possible implications in coffee breeding are discussed. [ABSTRACT FROM AUTHOR]

Introduction: Plants have been used to treat various ailments and diseases, including viral infections. Often activity is reported after screening plants traditionally used, without identifying the active principles.Objectives: This study investigated the use of metabolomics to identify common compound groups or compounds from unrelated plants, but with similar reported biological activity. Plants with anti-viral activities against Herpes Simplex Virus (HSV), Cytomegalovirus (CMV) and Human Immunodeficiency Virus (HIV) were collected and analysed. A few non-active plants, with no reported anti-viral activity were included as control samples.Methods: 1H-NMR and LC-MS metabolomic analysis were conducted, to determine the chemical similarity between plants with similar activity using SIMCA and XCMS online.Results: Plants with anti-HSV, anti-HIV and anti-CMV activity, presented specific clusters, with the non-active samples separating from the active samples. The anti-HSV group presented a clear contribution plot and chlorogenic acid was identified by NMR. LC-MS metabolomic analysis confirmed the NMR results and furthermore identified several chlorogenic acid isomers including the main substructures of chlorogenic acids.Conclusion: Metabolomic analysis on unrelated plants with similar activity can be used to identify the active compound groups or compounds, thereby eliminating the need for screening of plants to determine biological activity, additionally providing information on possible active principles. The two analytical methods identified chlorogenic acids and its building blocks as common and important compounds within plants with anti-HSV activity. Intensified research on plants containing chlorogenic acids should be the focus of future research for development of accessible anti-HSV treatments. [ABSTRACT FROM AUTHOR]