Isolation, identification, and quantification of roasted coffee antibacterial compounds

Coffee brew is a widely consumed beverage with multiple biological activities due both to naturally occurring components and to the hundreds of chemicals that are formed during the roasting process. Roasted coffee extract possesses antibacterial activity against a wide range of microorganisms, including Staphylococcus aureus and Streptococcus mutans, whereas green coffee extract exhibits no such activity. The naturally occurring coffee compounds, such as chlorogenic acids and caffeine, cannot therefore be responsible for the significant antibacterial activity exerted by coffee beverages against both bacteria. The very low minimum inhibitory concentration (MIC) found for standard glyoxal, methylglyoxal, and diacetyl compounds formed during the roasting process points to these alpha-dicarbonyl compounds as the main agents responsible for the antibacterial activity of brewed coffee against Sa. aureus and St. mutans. However, their low concentrations determined in the beverage account for only 50% of its antibacterial activity. The addition of caffeine, which has weak intrinsic antibacterial activity, to a mixture of alpha-dicarbonyl compounds at the concentrations found in coffee demonstrated that caffeine synergistically enhances the antibacterial activity of alpha-dicarbonyl compounds and that glyoxal, methylglyoxal, and diacetyl in the presence of caffeine account for the whole antibacterial activity of roasted coffee.     

Studies of selenium-containing volatiles in roasted coffee

Coffee has been an important and heavily used beverage in many cultures over a long period of time. Although sulfur species have been found to be abundant constituents, no work to date has explored the presence of selenium analogues. Investigation of volatile selenium species from green coffee beans, roasted beans, and brewed coffee drink was performed using solid phase microextraction (SPME) sample preconcentration in conjunction with GC/ICP-MS. Several volatile selenium species at trace levels were detected from roasted coffee beans as well as in the steam from brewed coffee drinks. No detectable selenium (and sulfur) species, however, were found in the headspace of green beans, indicating that selenium-containing volatiles are formed during roasting, as is the case for the sulfur volatiles. Matching standards were prepared and used to identify the compounds found in coffee. Artificial supplementation of the green coffee beans with selenium before roasting was performed to further characterize the selenium-containing volatiles formed during the coffee-roasting process.     

Roasting and aroma formation: effect of initial moisture content and steam treatment

Initial moisture of green coffee may vary as a function of green coffee processing and storage conditions. The impact of initial moisture and steam treatment on roasting behavior and aroma formation was investigated. Steam treated coffees as well as coffees with initial moisture content of 5.10, 10.04, and 14.70 g water per 100 g wb were roasted. Light and dark roasting trials were carried out using a fluidizing-bed roaster with a batch size of 100 g of green beans. Differences in roast coffee attributes, that is, color, density, and organic roast loss, and odorant concentrations were more marked in light roasted than in dark roasted coffees. The results of roasting steam treated coffee suggest that this step affects roasting behavior primarily by extracting some aroma precursor compounds.     

Effect of roasting conditions on the polycyclic aromatic hydrocarbon content in ground Arabica coffee and coffee brew

Roasting is a critical process in coffee production as it enables the development of flavor and aroma. At the same time, roasting may lead to the formation of nondesirable compounds, such as polycyclic aromatic hydrocarbons (PAHs). In this study, Arabica green coffee beans from Cuba were roasted under controlled conditions to monitor PAH formation during the roasting process. Roasting was performed in a pilot spouted bed roaster, with the inlet air temperature varying from 180 to 260 degrees C, using both dark (20 min) and light (5 min) roasting conditions. Several PAHs were determined in both roasted coffee samples and green coffee samples. Also, coffee brews, obtained using an electric coffee maker, were analyzed for final estimation of PAH transfer coefficients to the infusion. Formation of phenanthrene, anthracene, and benzo[a]anthracene in coffee beans was observed at temperatures above 220 degrees C, whereas formation of pyrene and chrysene required 260 degrees C. Low levels of benzo[g,h,i]perylene were also noted for dark roasting under 260 degrees C, with simultaneous partial degradation of three-cycle PAHs, suggesting that transformation of low molecular PAHs to high molecular PAHs occurs as the roasting degree is increased. The PAH transfer to the infusion was quite moderate (<35%), with a slightly lower extractability for dark-roasted coffee as compared to light-roasted coffee.     

Screening of raw coffee for thiol binding site precursors using "in bean" model roasting experiments

The purpose of the following study was to investigate the influence of coffee roasting on the thiol-binding activity of coffee beverages, and to investigate the potential of various green bean compounds as precursors of thiol-binding sites by using promising "in bean" model roast experiments. Headspace gas chromatographic analysis on coffee brews incubated in the presence of the roasty-sulfury smelling 2-furfurylthiol for 20 min at 30 degrees C in septum-closed vessels revealed that the amounts of "free" thiol decreased drastically with increasing the roasting degree of the beans used for preparation of the brews. A half-maximal binding capacity (BC(50)) of 183 mg of 2-furfurylthiol per liter of standard coffee beverage was determined for a roasted coffee (CTN value of 67), thus demonstrating that enormous amounts of the odor-active thiol are "bound" by the coffee. Furthermore, biomimetic "in bean" precursor experiments have been performed in order to elucidate the precursor for the thiol-binding sites in the raw coffee bean. These experiments opened the possibility of studying coffee model reactions under quasi-natural roasting conditions and undoubtedly identified chlorogenic acids as well as thermal degradation products caffeic acid and quinic acid as important precursors for low-molecular-weight thiol-binding sites. In particular, when roasted in the presence of transition metal ions, chlorogenic acids and even more caffeic acid showed thiol-binding activity which was comparable to the activity measured for the authentic coffee brew.     

Incidence,level, and behavior of aflatoxins during coffee bean roasting and decaffeination

Screening for aflatoxins (Afs), isolation and identification of Aspergillus flavus, and the effect of decaffeination and roasting on the level of contamination in coffee beans are studied. The percent frequency of A. flavus ranged between 4 and 80% in green coffee beans (GCB), whereas in ground roasted coffee beans (GRCB), it ranged between 1 and 71%. Aflatoxins were detected in 76.5 and 54.6% of the infected samples with averages of 4.28 and 2.85 microg/kg of GCB and GRCB, respectively. Roasting was demonstrated to lower the concentration of Afs in GCB. The Afs levels were reduced by approximately 42.2-55.9% depending on the type and temperature of roasting. The highest yields of Afs were detected in the decaffeinated green coffee beans (24.29 microg/kg) and roasted coffee beans (16.00 microg/kg). The growth of A. flavus in liquid medium containing 1 or 2% caffeine was reduced by 50%, and the level of aflatoxin in the medium was undetectable.     

Formation of flavor components by the reaction of amino acid and carbonyl compounds in mild conditions

This work describe products of reactions between four alpha-dicarbonyl compounds (diacetyl, pentan-2,3-dione, glyoxal, and methylglyoxal) or two alpha-hydroxy ketones, (acetoine and acetol) and amino acids present in wines. The results shows the formation of odorous products or strong-smelling additives resulting from the Maillard and Strecker reaction in a primarily aqueous medium, at low temperature and low pH ( approximately pH 3.5) of the wine. GC/FID, GC/FPD, GC/NPD and GC/MS techniques were used. The olfactive characteristics of the products are described. In the presence of sulfur amino acids and in particular cysteine, many products were formed with a heterocycle production such as pyrazines and methylpyrazines, methylthiazoles, acetylthiazoles, acetylthiazolines, acetylthiazolidines, trimethyloxazole, and dimethylethyloxazoles. These various compounds present odors of sulfur, cornlike, pungent, nut, popcorn, roasted hazelnut, toasted, roasted, and ripe fruits. The chemical conditions of the model reactions are specified. The influence of temperature and pH on the reactions in the presence of cysteine were also studied.     

Modeling the formation of some polycyclic aromatic hydrocarbons during the roasting of Arabica coffee samples

Roasting is a critical process in coffee production, as it enables the development of flavor and aroma. At the same time, roasting may lead to the formation of nondesirable compounds, such as polycyclic aromatic hydrocarbons (PAHs). In this study, Arabica green coffee beans from Cuba were roasted under controlled conditions to monitor PAH formation during the roasting process. Roasting was performed in a pilot-spouted bed roaster, with the inlet air temperature varying from 180 to 260 degrees C, for roasting conditions ranging from 5 to 20 min. Several PAHs were determined in both roasted coffee samples and green coffee samples. Different models were tested, with more or less assumptions on the chemical phenomena, with a view to predict the system global behavior. Two kinds of models were used and compared: kinetic models (based on Arrhenius law) and statistical models (neural networks). The numbers of parameters to adjust differed for the tested models, varying from three to nine for the kinetic models and from five to 13 for the neural networks. Interesting results are presented, with satisfactory correlations between experimental and predicted concentrations for some PAHs, such as pyrene, benz[a]anthracene, chrysene, and anthracene.     

Effect of roasting on the antioxidant activity of coffee brews

Colombian Arabica coffee beans were roasted to give light, medium, and dark samples. Their aqueous extracts were analyzed by gel filtration chromatography, UV-visible spectrophotometry, capillary electrophoresis, and the ABTS(*)(+) assay. A progressive decrease in antioxidant activity (associated mainly with chlorogenic acids in the green beans) with degree of roasting was observed with the simultaneous generation of high (HMM) and low molecular mass (LMM) compounds possessing antioxidant activity. Maximum antioxidant activity was observed for the medium-roasted coffee; the dark coffee had a lower antioxidant activity despite the increase in color. Analysis of the gel filtration chromatography fractions showed that the LMM fraction made a greater contribution to total antioxidant activity than the HMM components.     

Interactions between volatile and nonvolatile coffee components. 1. Screening of nonvolatile components

This study is the first of two publications that investigate the phenomena of coffee nonvolatiles interacting with coffee volatile compounds. The purpose was to identify which coffee nonvolatile(s) are responsible for the interactions observed between nonvolatile coffee brew constituents and thiols, sulfides, pyrroles, and diketones. The overall interaction of these compounds with coffee brews prepared with green coffee beans roasted at three different roasting levels (light, medium, and dark), purified nonvolatiles, and medium roasted coffee brew fractions (1% solids after 1 or 24 h) was measured using a headspace solid-phase microextraction technique. The dark roasted coffee brew was slightly more reactive toward the selected compounds than the light roasted coffee brew. Selected pure coffee constituents, such as caffeine, trigonelline, arabinogalactans, chlorogenic acid, and caffeic acid, showed few interactions with the coffee volatiles. Upon fractionation of medium roasted coffee brew by solid-phase extraction, dialysis, size exclusion chromatography, or anion exchange chromatography, characterization of each fraction, evaluation of the interactions with the aromas, and correlation between the chemical composition of the fractions and the magnitude of the interactions, the following general conclusions were drawn. (1) Low molecular weight and positively charged melanoidins present significant interactions. (2) Strong correlations were shown between the melanoidin and protein/peptide content, on one hand, and the extent of interactions, on the other hand (R = 0.83-0.98, depending on the volatile compound). (3) Chlorogenic acids and carbohydrates play a secondary role, because only weak correlations with the interactions were found in complex matrixes.     

In vitro antioxidant and ex vivo protective activities of green and roasted coffee

The antioxidant properties of green and roasted coffee, in relation to species (Coffea arabica and Coffea robusta) and degree of roasting (light, medium, dark), were investigated. These properties were evaluated by determining the reducing substances (RS) of coffee and its antioxidant activity (AA) in vitro (model system beta-carotene-linoleic acid) and ex vivo as protective activity (PA) against rat liver cell microsome lipid peroxidation measured as TBA-reacting substances. RS of C. robustasamples were found to be significantly higher when compared to those of C. arabica samples (p < 0.001). AA for green coffee samples were slightly higher than for the corresponding roasted samples while PA was significantly lower in green coffee compared to that of all roasted samples (p < 0.001). Extraction with three different organic solvents (ethyl acetate, ethyl ether, and dichloromethane) showed that the most protective compounds are extracted from acidified dark roasted coffee solutions with ethyl acetate. The analysis of acidic extract by gel filtration chromatography (GFC) gave five fractions. Higher molecular mass fractions were found to possess antioxidant activity while the lower molecular mass fractions showed protective activity. The small amounts of these acidic, low molecular mass protective fractions isolated indicate that they contain very strong protective agents.     

Fourier transform infrared and physicochemical analyses of roasted coffee

In this study, Brazilian coffee beans processed to different stages of roast at 210, 220, 230, and 240 °C were analyzed for pH value, titratable acidity, moisture content, and color lightness. Fourier transform infrared (FTIR) spectroscopy, in conjunction with principal component analysis, was conducted to study the effects of process time and temperature on the IR-active components of the acetyl acetate extract of the roasted coffee. The results showed that high-temperature-short-time resulted in higher moisture content, higher pH value, and higher titratable acidity when the beans were roasted beyond the start-of-second-crack stage, as compare to low-temperature-long-time process (LTLT). The LTLT process also resulted in greater IR absorbance for aldehydes, ketones, aliphatic acids, aromatic acids, and caffeine carbonyl bands on the FTIR spectra. Clusters for principal component score plots were well separated, indicating that the changes IR-active components in the coffee extracts, due to the different roasting treatments, can be discriminated by the FTIR technique. On the basis of the loading plots of principal components, changes of IR-active compounds in the coffee extract at various stages of roasting were discussed.     

Insight into the Mechanism of Coffee Melanoidin Formation Using Modified "in Bean" Models

To study the mechanism of coffee melanoidin formation, green coffee beans were prepared by (1) removal of the hot water extractable components (WECoffee); (2) direct incorporation of sucrose (SucCoffee); and (3) direct incorporation of type II arabinogalactan-proteins (AGPCoffee). As a control of sucrose and AGP incorporation, lyophilized green coffee beans were also immersed in water (control). The original coffee and the four modified "in bean" coffee models were roasted and their chemical characteristics compared. The formation of material not identified as carbohydrates or protein, usually referred to as "unknown material" and related to melanoidins, and the development of the brown color during coffee roasting have distinct origins. Therefore, a new parameter for coffee melanoidin evaluation, named the "melanoidin browning index" (MBI), was introduced to handle simultaneously the two concepts. Sucrose is important for the formation of colored structures but not to the formation of "unknown material". Type II AGPs also increase the brown color of the melanoidins, but did not increase the amount of "unknown material". The green coffee hot water extractable components are essential for coffee melanoidin formation during roasting. The cell wall material was able to generate a large amount of "unknown material". The galactomannans modified by the roasting and the melanoidin populations enriched in galactomannans accounted for 47% of the high molecular weight brown color material, showing that these polysaccharides are very relevant for coffee melanoidin formation.     

Chlorogenic acids and lactones in regular and water-decaffeinated arabica coffees

The market for decaffeinated coffees has been increasingly expanding over the years. Caffeine extraction may result in losses of other compounds such as chlorogenic acids (CGA) and, consequently, their 1,5-gamma-quinolactones (CGL) in roasted coffee. These phenolic compounds are important for flavor formation as well as the health effects of coffee; therefore, losses due to decaffeination need to be investigated. The present study evaluates the impact of decaffeination processing on CGA and CGL levels of green and roasted arabica coffees. Decaffeination produced a 16% average increase in the levels of total CGA in green coffee (dry matter), along with a 237% increase in CGL direct precursors. Different degrees of roasting showed average increments of 5.5-18% in CGL levels of decaffeinated coffee, compared to regular, a change more consistent with observed levels of total CGA than with those of CGL direct precursors in green samples. On the other hand, CGA levels in roasted coffee were 3-9% lower in decaffeinated coffee compared to regular coffee. Although differences in CGA and CGL contents of regular and decaffeinated roasted coffees appear to be relatively small, they may be enough to affect flavor characteristics as well as the biopharmacological properties of the final beverage, suggesting the need for further study.     

Identification of odor-active 3-mercapto-3-methylbutyl acetate in volatile fraction of roasted coffee brew isolated by steam distillation under reduced pressure

In a roasted Arabica coffee brew, the potent roasty odor quality compound was identified as 3-mercapto-3-methylbutyl acetate by comparison of its Kovats gas chromatography retention index, mass spectrum, and odor quality to those of the synthetic authentic compound. 3-Mercapto-3-methylbutyl acetate has been identified for the first time in the coffee, and according to the results of the aroma extract dilution analysis, the contribution of this compound to the flavor of the roasted coffee brew varied depending on the degree of the coffee bean roasting. The concentration of this compound in the coffee brews as with 3-mercapto-3-methylbutyl formate increased with an increase in the degree of roasting. However, the slope of the amount of both esters was different, and 3-mercapto-3-methylbutyl acetate hardly increased with a low degree of roasting at more than a 21 luminosity (L)-value, but it rapidly increased when the roasting degree of the coffee beans reached the L-value of 18. These results suggested that the contribution of 3-mercapto-3-methylbutyl acetate to the overall flavor is peculiar to the flavor of the highly roasted coffee.     

Evaluation of methods used to determine ochratoxin A in coffee beans

A comparative study was conducted to evaluate four previously reported methods that proved to have a recovery greater than 80% for the determination of different levels of ochratoxin A (OTA) in green and roasted coffee beans and to select an accurate, sensitive, and less-expensive technique between the existing methods. The results indicated that the Association of Official Analytical Chemists (AOAC) official method for the extraction of OTA in green coffee and determination by high-performance liquid chromatography (HPLC) is recommended as an efficient method for the routine analyses of OTA in green and ground roasted coffee beans. This method proved to be an accurate, sensitive, and less-expensive method that employs routine materials and available equipment. Although the immunoaffinity column/HPLC procedure tested showed a significantly higher percentage than the AOAC recommended method, it is recommended for use in processed coffee beans where low concentrations of OTA may be expected to be detected.     

Analysis of volatile compounds released during the grinding of roasted coffee beans using solid-phase microextraction

A dynamic solid-phase microextraction (SPME) method to sample fresh headspace volatile compounds released during the grinding of roasted coffee beans was described and the analytical results using gas chromatography/mass spectrometry (GC/MS) and GC/olfactometry (GC/O) were compared to those of the conventional static SPME sampling methods using ground coffee. Volatile compounds released during the grinding of roasted coffee beans (150 g) were obtained by exposing the SPME fiber (poly(dimethylsiloxane)/divinylbenzene, PDMS/ DVB) for 8 min to nitrogen gas (600 mL/min) discharged from a glass vessel in which the electronic coffee grinder was enclosed. Identification and characterization of volatile compounds thus obtained were achieved by GC/MS and GC/O. Peak areas of 47 typical coffee volatile compounds, separated on total ion chromatogram (TIC), obtained by the dynamic SPME method, showed coefficients of variation less than 5% (n = 3) and the gas chromatographic profile of volatile compounds thus obtained was similar to that of the solvent extract of ground coffee, except for highly volatile compounds such as 4-hydroxy-2,5-dimethyl-3(2H)-furanone and 4-ethenyl-2-methoxyphenol. Also, SPME dilution analysis of volatile compounds released during the grinding of roasted coffee beans showed linear plots of peak area versus exposed fiber length (R (2) > 0.89). Compared with those of the headspace volatile compounds of ground coffee using GC/MS and GC/O, the volatile compounds generated during the grinding of roasted coffee beans were rich in nutty- and smoke-roast aromas.     

Coffee roasting and aroma formation: application of different time-temperature conditions

The impact of time-temperature combinations of roasting processes on the kinetics of aroma formation in coffee was investigated. The development of 16 aroma compounds and the physical properties of coffee beans was followed in a commercial horizontal drum roasting process and in laboratory scale fluidizing-bed roasting processes at high temperature-short time and low temperature-long time conditions. All trials were run to an equal roast end point as defined by the lightness of coffee beans. In addition, the effect of excessive roasting on aroma composition was studied. Compared to low temperature-long time roasting, high temperature-short time roasting resulted in considerable differences in the physical properties and kinetics of aroma formation. Excessive roasting generally led to decreasing or stable amounts of volatile substances, except for hexanal, pyridine, and dimethyl trisulfide, whose concentrations continued to increase during over-roasting. When the drum roaster and the fluidizing bed roaster were operated in the so-called temperature profile mode, that is, along the identical development of coffee bean temperature over roasting time, the kinetics of aroma generation were similar in both processes.     

Furan levels in coffee as influenced by species, roast degree, and brewing procedures

Brazilian green coffee beans of Coffea arabica and Coffea canephora species were roasted to light, medium, and dark roast degrees and analyzed in relation to furan content by using an in-house validated method based on gas chromatography coupled to mass spectrometry preceded by headspace solid-phase microextraction. Furan was not detected in green coffees, whereas levels between 911 and 5852 μg/kg were found in the roasted samples. Higher concentrations were found in Coffea canephora species and darker ground coffees. Some of the potential furan precursors were observed in significant amounts in green coffee, especially sucrose and linoleic acid, but their concentrations could not be correlated to furan formation. Additionally, coffee brews were prepared from roasted ground coffees by using two different procedures, and furan levels in the beverages varied from <10 to 288 μg/kg. The factor that most influenced the furan content in coffee brew was the brewing procedure.