Chlorogenic Acid Isomers Research Articles

Changes of the chlorogenic acid, caffeine, gama-aminobutyric acid (GABA) and antioxdant activities during germination of coffee bean (Coffea arabica)

This study was carried out to identify changes in functional materials during the germination of coffee seeds. Chlorogenic acid (CGA), caffeine, and g-aminobutyric acid (GABA) was assayed by HPLC method while germinating coffee beans for 20 days. The contents of total phenolic compounds and total flavonoid compounds were measured by colorimetric method. Antioxidant activity was measured by both 2,2-diphenyl-2-pycryl hydrazyl (DPPH) radical scavenging activity assay and 2,2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS•+) assay method. It was found that CGAs (5-caffeoylquinic acid (CQA), 4-CQA, 3-CQA), caffeine, and GABA increased and then decreased during germination. There were no significant changes in total phenolics, total flavonoids and antioxidant activities. Especially, all of the major isomers of CGA increased to the highest value at 10 days of germination, and GABA also increased significantly at the early stage of germination. This study showed that coffee beans changed into more functional food materials by germination and showed maximum functionality in appropriate germination conditions.

UHPLC-ESI-QqTOF-MS/MS characterization of minor chlorogenic acids in roasted Coffea arabica from different geographical origin.

Chlorogenic acids are relevant coffee quality markers, taste, and aroma precursors as well as important bioactive compounds. A number of mono-acyl, di-acyl, and tri-acyl quinic acid isomers were found in green coffee beans, being mono-caffeoyl, mono-feruloyl, mono-p-coumaroyl, and di-caffeoylquinic acid isomers considered as quantitatively major compounds. Roasting process increases the chemical complexity of coffee by inducing the formation of a number of lactones (quinides), shikimates, and other chlorogenic acids derivatives. So far, little attention has been paid in characterizing minor chlorogenic acids and derivatives in roasted Coffea arabica, also known as Arabica. In the present work, roasted C. arabica samples from different geographical origins (Brazil, Colombia, Costa Rica, Ethiopia, Guatemala, and India) were characterized by UHPLC-ESI-QqTOF-MS/MS. Several minor chlorogenic acid isomers were identified. In particular, HR-MS/MS provided putative identification of four dimethoxycinnamoyl-quinic acid derivatives, such as 4-dimethoxycinnamoylquinic acid, 4-dimethoxycinnamoyl-3-caffeoylquinic acid, 3-dimethoxycinnamoyl-4-feruloylquinic acid, 4-dimethoxycinnamoyl-5-feruloylquinic acid, and two caffeoyl, feruloyl quinic acid derivatives (3-caffeoyl-4-feruloylquinic acid and 3-feruloyl-4-caffeoylquinic acid). To our knowledge, these compounds were found in roasted Arabica coffee for the first time, and their presence is independent on the different geographical origins examined.

Analysis of chlorogenic acids isomers and caffeic acid in 89 herbal infusions (tea)

Several plants used to prepare aqueous beverages (teas) are known for their health benefits due to the presence of bioactive compounds, such as isomers of 3-caffeoylquinic (3-CQ), 4-caffeoylquinic (4-CQ), 5-caffeoylquinic (5-CQ), 3,4-dicaffeoylquinic (3,4-DQ), 3,5-dicaffeoylquinic (3,5-DQ), 4,5-dicaffeoylquinic (4,5-DQ) and caffeic acid. This study analyses the presence of these compounds in infusions from 89 plants. We acquired these plants from public markets and analyzed them using ultra-performance liquid chromatography. We found the listed chlorogenic acids in 93% of infusions. The infusions containing highest concentrations were yerba mate (Ilex paraguariensis), with 52.6 mg in 100 mL of infusion, followed by assa-peixe (Boehmeria caudata), white tea (Camellia sinensis), winter’s bark (Drimys winteri), green tea (Camellia sinensis), and elderflower (Sambucus nigra), with 6.0 mg in 100 mL of infusion. Several plant infusions had not been previously studied. Compared to other foods and beverages, these infusions are considered potential sources of chlorogenic acids.

From the field to coffee cup: impact of planting design on chlorogenic acid isomers and other compounds in coffee beans and sensory attributes of coffee beverage

Various chemical compounds in green coffee beans are precursors of coffee beverage sensory attributes. The hypothesis was that those chemical compounds would vary with microclimate over the vertical plant profile of coffee plants. The plants of the cultivar IAPAR 59 were grown in two high plant densities (6000 and 10,000 plants ha−1) under rectangular and square planting patterns. Coffee beans were collected from three canopy strata. Chemical compounds and chlorogenic acid isomer (CGA) contents were determined by near infrared spectroscopy and ultra-performance liquid chromatography, respectively. The free-choice profile technique was employed to estimate the sensory attributes of coffee beverage. Brightness, transparency appearance, typical coffee color and aroma, tastes as sweet, acid, astringent and bitter, and full-bodied texture were attributes detected by panelists. Plants cultivated under lower plant density had lower leaf area index indicating lower self-shading than under higher plant density. Coffee beans originated from the inferior stratum of higher density had the lowest concentrations of 5-caffeoylquinic acid (5-CQA), while the inverse situation occurred in their upper strata. Strata of lower planting density were associated to high levels of all CGA isomers. The coffees cultivated under higher density showed high lipids and CGA isomers contents. Under lower density, beans had low concentration of sucrose, proteins, and caffeine and high of CGA isomer contents. The beverage sensory attributes depended on the concentration of proteins, lipids, and sucrose in combination to CQA isomers in beans, varying over the vertical profile and planting designs of coffee plants.

Site-specific hydrolysis of chlorogenic acids by selected Lactobacillus species

Hydroxycinnamic acids are a major group of phenolic compounds widely distributed in plants. Among them, chlorogenic acids and caffeic acid have been in the focus of interest due to their impact on food quality and their putative health benefits. Numerous microorganisms like lactic acid bacteria are able to hydrolyze chlorogenic acids by cinnamoyl esterase enzymes. Data on the specificity of theses enzymes regarding the cleavage of distinct isomers of mono- or dichlorogenic acids is lacking. Lactobacillus reuteri, Lactobacillus helveticus, and Lactobacillus fermentum were screened for their ability to hydrolyze chlorogenic acid isomers in culture medium. Concentrations of chlorogenic acids and the released caffeic acid were determined by UHPLC-ESI-MS. The highest hydrolysis rate (100%) was observed for the hydrolysis of 5-CQA by Lactobacillus helveticus. A so far unknown metabolic pathway for the cleavage of 4-CQA is proposed including isomerization to 5-CQA and 3-CQA followed by hydrolysis.

Roasting has a distinct effect on the antimutagenic activity of coffee varieties

Coffee is a highly consumed beverage throughout the world. Its popularity derives from its organoleptic properties that are a result of the roasting process. Roasting greatly alters a coffee bean’s composition and possibly its bioactivity. In the current study, green as well as roasted extracts from both Coffea arabica (Brazil and Decaf) and Coffea canephora (Robusta) species were tested for their antimutagenic activity using the Ames test. In addition, a compositional analysis was conducted to identify the main components, mainly Chlorogenic acid isomers (CGA) and derivatives present in the extracts using UHPLC-ESI(±) and HRMS/MS methods According to the results, all extracts exhibited strong antimutagenic activity against the oxidizing factor tert-Butyl hydroperoxide, a Reactive Oxygen Species-producing compound. Roasting had a distinct effect on the antimutagenic activity of coffee, enhancing it in the Brazil variety and having no effect in the Decaf and Robusta varieties. In addition, all coffee extracts exhibited reducing activity as well as the ability to scavenge (albeit differentially) both the superoxide and hydroxyl radicals, implying that their potential antimutagenic effect can be partially attributed to their free radical scavenging activity.

Roasted and green coffee extracts show antioxidant and cytotoxic activity in myoblast and endothelial cell lines in a cell specific manner

Coffee is one of the most highly consumed beverages with potential beneficial health implications, however its molecular mechanism of action has not been completely elucidated yet. To that cause, the polyphenolic composition of different coffee extracts (from Light, Medium and Dark roasts as well as green beans) was examined by UHPLC-HRMS analysis, indicating chlorogenic acids isomers as the main constituents. In the following step, the toxicity of the extracts was tested in myoblasts and endothelial cells and differential toxicity of green and roasted samples was displayed as the myoblasts were more sensitive to green coffee extracts, in contrast to the endothelial cells. Subsequently, biologically relevant, non-cytotoxic extract concentrations were administered to explore their potential effect on cell redox status using flow cytometry and spectrophotometric assays. The results indicated that all coffee extracts improved cell redox status, however differences were observed between the two different cell lines tested, implying that coffee compounds display cell- and tissue-specificity. Glutathione levels were increased in almost all cases up to 70%, while the roasting degree affected the free radical scavenging potential of the extracts and their ability to protect from macromolecular oxidation as exhibited by the differences in ROS, CARB and TBARS levels, especially in the myoblasts.

Chromatographic and Voltammetric Characterization of Chlorogenic Acids in Coffee Samples

The accurate and precise characterization of eight chlorogenic acids (CGAs) isomers, caffeoylquinic acids (5-CQA, 4-CQA, 3-CQA), dicaffeoylquinic acids (3, 5-diCQA, 4, 5-diCQA, 3, 4-diCQA), and feruloylquinic acids (5-FQA, 4- FQA), was carried out using different brands of coffee. High-performance liquid chromatography (HPLC), differential pulse (DPV) and square- wave voltammetry (SWV) were applied for the investigation. HPLC proved to be an accurate and precise method for separation, identification and quantification of CGAs isomers in coffee extracts. DPV and SWV have shown that electrochemical behaviour of coffee extracts was very similar to that of CGAs isomers and DPV and SWV can be used for the correct characterization of CGAs in coffee. All three techniques have shown a very high correlation of CGAs content in all investigated coffee samples. Therefore, HPLC, DPV and SWV methods can be used in combination as very selective, sensitive and precise methods for characterization of CGAs isomers in coffee extracts.

Morphological characteristics, chemical and genetic diversity of kenaf (Hibiscus cannabinusL.) genotypes

Jaihyunk Ryu, Soon-Jae Kwon, Dong-Gun Kim, Min-Kyu Lee, Jung Min Kim, Yeong Deuk Jo, Sang Hoon Kim, Sang Wook Jeong, Kyung-Yun Kang, Se Won Kim, Jin-Baek Kim, and Si-Yong Kang. J Plant Biotechnol 2017;44:416-30. https://doi.org/10.5010/JPB.2017.44.4.416

Determination of chlorogenic acids and caffeine in homemade brewed coffee prepared under various conditions.

Coffee, a complex mixture of more than 800 volatile compounds, is one of the most valuable commodity in the world, whereas caffeine and chlorogenic acids (CGAs) are the most common compounds. CGAs are mainly composed of caffeoylquinic acids (CQAs), dicaffeoylquinic acids (diCQAs), and feruloylquinic acids (FQAs). The major CGAs in coffee are neochlorogenic acid (3-CQA), cryptochlorogenic acid (4-CQA), and chlorogenic acid (5-CQA). Many studies have shown that it is possible to separate the isomers of FQAs by high-performance liquid chromatography (HPLC). However, some authors have shown that it is not possible to separate 4-feruloylquinic acid (4-FQA) and 5-feruloylquinic acid (5-FQA) by HPLC. Therefore, the present study was designated to investigate the chromatographic problems in the determination of CGAs (seven isomers) and caffeine using HPLC-DAD. The values of determination coefficient (R2) calculated from external-standard calibration curves were >0.998. The recovery rates conducted at 3 spiking levels ranged from 99.4% to 106.5% for the CGAs and from 98.8% to 107.1% for the caffeine. The precision values (expressed as relative standard deviations (RSDs)) were <7% and <3% for intra and interday variability, respectively. The tested procedure proved to be robust. The seven CGAs isomers except 4-FQA and 5-FQA were well distinguished and all gave good peak shapes. We have found that 4-FQA and 5-FQA could not be separated using HPLC. The method was extended to investigate the effects of different brewing conditions such as the roasting degree of green coffee bean, coffee-ground size, and numbers of boiling-water pours, on the concentration of CGAs and caffeine in homemade brewed coffee, using nine green coffee bean samples of different origins. It was reported that medium-roasted, fine-ground coffees brewed using three pours of boiling water were the healthiest coffee with fluent CGAs.

Characterization of Chlorogenic Acids in Coffee by Flow-Through Chronopotentiometry

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 −OCH3 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−1 with the limit of detection 5.7·10−7 mol L−1. 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.

Chlorogenic acid isomer contents in 100 plants commercialized in Brazil

This study analysed 100 plants employed in Brazil as ingredients to infusions for their caffeic acid, 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) contents. The samples were collected from public markets and analysed using ultra-high performance liquid chromatography (UPLC). The highest concentrations of chlorogenic acids were found in yerba mate (Ilex paraguariensis), 9,2g·100g−1, white tea (Camellia sinensis), winter's bark (Drimys winteri), green tea (Camellia sinensis), elderflower (Sambucus nigra), and Boehmeria caudata (known as assa-peixe in Brazil), 1,1g·100g−1. The present work showcased the investigation of chlorogenic acids in a wide range of plants not yet studied in this regard and also resulted in a comparative table which explores the content of six isomers in the samples.

Polyphenol patterns to trace sweet (Prunus avium) and tart (Prunus cerasus) varieties in cherry jam.

The aim of this work was to assess whether the characteristic polyphenol traits of cherry biotypes persisted in thermally processed cherry products, such as jam. Thus, the RP-HPLC-diode array detector profiles of both colorless polyphenols and anthocyanins from three cherry varieties (two sweet and one tart cherry) were compared with those of low-sugar jam sourced from the same cultivars. Individual components were characterized by mass spectrometry. The total phenolic and total anthocyanin content as well as the radical scavenging potential (residual 75-91, 88-91 and 73-75%, respectively) were only slightly reduced by deep thermal treatments. Apart from the interconversion among the isomers of chlorogenic acid, the profile of both colorless polyphenols and anthocyanins substantially survived the jam manufacturing under conventional temperature-time regimen (80°C, 1h). The species- and cultivar-specific polyphenol molecular asset, especially the anthocyanin pattern, has potential to be monitored for traceability purpose, aimed to the varietal assessment of cherry biotypes used for producing jam.

Glycoalkaloid, phenolic, and flavonoid content and antioxidative activities of conventional nonorganic and organic potato peel powders from commercial gold, red, and Russet potatoes

Potato peels from the large-scale peeling of potatoes are a source of bioactive compounds. We determined the content of the glycoalkaloids α-chaconine and α-solanine, individual and total phenolics compounds, and total flavonoids extracted from three peels prepared from conventional and three from organic commercial gold, red, and Russet potatoes as well as determining their antioxidant activities using three different methods. Glycoalkaloids and individual phenolic compounds (caffeic acid, chlorogenic acid, and chlorogenic acid isomer) were determined using HPLC and LC–MS, and total phenolic and flavonoid content by colorimetry. There was a wide variation in the content of total glycoalkaloids, phenolic compounds, flavonoids, and antioxidative activities, as indicated by the following ratios for the total organic to nonorganic glycoalkaloids: gold, 3.89; Russet, 2.43; and red, 0.50. The corresponding phenolics and flavonoid ratios indicated a narrower range. The described methods could be used to determine the composition and antioxidative activities of peels derived from other potato cultivars, taking into account their methods of growth and postharvest storage, and aid in compositional, nutritional, medical, and industrial studies on potato peel, to relate their composition to their many described applications.

A robust platform based on ultra-high performance liquid chromatography Quadrupole time of flight tandem mass spectrometry with a two-step data mining strategy in the investigation, classification, and identification of chlorogenic acids in Ainsliaea fragrans Champ

The chlorogenic acids are the major bioactive constituents of the whole plant of Ainsliaea fragrans Champ. (Xingxiang Tuerfeng). These compounds are usually present as isomeric forms in Xingxiang Tuerfeng. Therefore, an efficient approach should be developed for the rapid discovery and identification of chlorogenic acids isomers through the fragmentation pathway and rules. In this study, the collision induced dissociation tandem mass spectrometry (CID-MS/MS) fragmentation routes of chlorogenic acids were systematically investigated by UHPLC–QTOF–MS/MS in the negative ion mode using eight chlorogenic acids standards. As a result, diagnostic product ions for rapid discovery and classification of chlorogenic acids isomers were determined according to their MS/MS fragmentation patterns and intensity analysis. Based on these findings, a novel two-step data mining strategy was established. The first key step was to screen different kinds of substitution and the skeleton of the quinic acid using the characteristic product ions and neutral losses. The second key step was to screen and classify different types of chlorogenic acids using their diagnostic product ions. It was apply to the rapid investigation, classification, and identification of chlorogenic acids. And the same carbon skeletons from a complex extract of Ainsliaea fragrans Champ. were effectively identified. 88 constituents, including 14 chlorogenic acids types, were rapidly discovered and identified, and in particular, 12 types of chlorogenic acids, including p-CoQC, FQA, BQC, CQA-Glu, CFQA, p-Co-CQC, di-p-CoQC, BCQA, di-CQA-Glu, PCQA, tri-QCA, and P-di-CQA, were first discovered in Ainsliaea fragrans Champ. In conclusion, UHPLC-QTOF-MS/MS method together with a systematic two-step data mining strategy was established as a feasible, effective, and rational technique for analyzing chlorogenic acids. Additionally, this study laid a foundation for the study of the active substances and quality control of Ainsliaea fragrans Champ.

Multivariate Optimization of Chlorogenic Acid Extraction From Brazilian Coffee

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.

Variation in polyphenolic profile and in vitro antioxidant activity of eastern teaberry (Gaultheria procumbens L.) leaves following foliar development

Seasonal dynamics in the polyphenolic composition, antioxidant activity, and their relationships during plant development were evaluated for eastern teaberry (Gaultheria procumbens L.) leaves, a traditional herbal medicine of North American natives. With the complementary UHPLC-PDA-ESI-MS3, HPLC-PDA-fingerprint, Folin-Ciocalteau, and n-butanol/HCl assays of methanol-water (75:25, v/v) extracts, the dried leaf samples harvested monthly across the growing season under Polish climate conditions were found rich in structurally diverse polyphenols (149.2–210.7mg/g DW) including the dominating salicylates (64.6–107.5mg/g DW), proanthocyanidins (53.0–66.8mg/g DW), and flavonoids (17.3–25.3mg/g DW), and the accompanying chlorogenic acid isomers (2.4–4.4mg/g DW) and simple phenolic acids (0.9–1.1mg/g DW). Among 28 detected analytes, gaultherin (64.6–107.5mg/g DW), miquelianin (14.6–21.1mg/g DW), procyanidin A-type trimer (5.5–9.5mg/g DW), and (–)-epicatechin (5.8–7.8mg/g DW) were the most abundant. The phenolic levels and antioxidant activity parameters in the DPPH (EC50, 15.0–18.2μg DW/mL; 0.95–1.16mmol Trolox equivalents/g DW) and FRAP (2.3–3.4mmolFe 2+/g DW; 0.86–1.26mmol Trolox equivalents/g DW) assays showed parallel seasonal trends with maxima in September and October. As the subsequent correlation studies confirmed the determinative impact of polyphenols on the leaf antioxidant activity and its seasonal fluctuations, the Fall season could be recommended as optimal for harvesting the plant material for medicinal purposes and cost-effective production of natural health products.

울릉 마가목의 클로로겐산 이성체의 최종당화산물의 생성 저해 및 라디칼 소거 활성

Advanced glycation end product (AGE) formation and reactive oxygen species are potential therapeutic targets for the prevention of diabetic nephropathy and other pathogenic complications. Activity-guided isolation of an ethylacetate-soluble portion of 80% methanolic extract from fruits of Sorbus commixta of the Ulleung Island origin using AGE formation inhibition assay led to the isolation and identification of three caffeoylquinic acid derivatives of a previously known structure, 3-O-caffeoylquinic acid (neochlorogenic acid; 1), 4-O-caffeoylquinic acid (cryptochlorogenic acid; 2), and 5-O-caffeoylquinic acid (chlorogenic acid; 3). The structures of these compounds were confirmed by interpretation of nuclear magnetic resonance and mass spectrometry data. Among the isolates, the major metabolite, neochlorogenic acid (1) showed the most potent inhibitory effect against AGE formation with an IC50 value of 167.5±3.5 μM. Furthermore, all isolated chlorogenic acid isomers were evaluated for their radical scavenging activity against peroxynitrite, and structurally related isomers 1, 2, and 3 exhibited potent inhibitory effects in this radical scavenging assay. This result suggests that the monocaffeoyl quinic acid derivatives isolated from S. commixta might be beneficial for the regulation of diabetic complications and related diseases.

Inhibitory effect of chlorogenic acid on digestion of potato starch

The effect of the chlorogenic acid isomer 5-O-caffeoylquinic acid (5-CQA) on digestion of potato starch by porcine pancreatic alpha amylase (PPAA) was investigated using isolated starch and cooked potato tuber as substrates. In vitro digestion was performed on five varieties of potato with varying phenolic content. Co- and pre-incubation of PPAA with 5-CQA significantly reduced PPAA activity in a dose dependent manner with an IC50 value of about 2mgmL−1. Lineweaver-Burk plots indicated that 5-CQA exerts a mixed type inhibition as km increased and Vmax decreased. The total polyphenol content (TPC) of peeled tuber tissue ranged from 320.59 to 528.94mg 100g−1dry weight (DW) in raw tubers and 282.03–543.96mg 100g−1DW in cooked tubers. With the exception of Désirée, TPC and 5-CQA levels decreased after cooking. Principle component analysis indicated that digestibility is affected by multiple factors including phenolic, dry matter and starch content.

Transformation of chlorogenic acids during the coffee beans roasting process

Due to the recently discovered biomedical activity of chlorogen acid, the interest in its physicochemical and pharmaceutical properties has increased. The plant product most frequently mentioned as remarkably rich in this compound is coffee beans. The present paper shows that the roasting process of coffee beans not only leads to the degradation of chlorogenic acid isomers to volatile components, the fact well known from the literature, but also causes their transformation to quinic acid and its epimer and to four chlorogenic acid lactones (basic and epimeric forms of 3- and 4-O-caffeoylquinic acid lactones) and changes the ratios between the amounts of chlorogen acid isomers and dicaffeoylquinic acids isomers. It is also shown that the increase of water content above its standard level in the initial green coffee beans slows down only the changes occurring during the roasting process.