Coffee Matrix Research Articles

Influence of aroma binding factors of coffee matrix on characteristic aroma in a model system

Influence of aroma binding factors of coffee matrix on characteristic aroma in a model system

Exploring the link between coffee matrix microstructure and flow properties using combined X-ray microtomography and smoothed particle hydrodynamics simulations

Coffee extraction involves many complex physical and transport processes extremely difficult to model. Among the many factors that will affect the final quality of coffee, the microstructure of the coffee matrix is one of the most critical ones. In this article, we use X-ray micro-computed (microCT) technique to capture the microscopic details of coffee matrices at particle-level and perform fluid dynamics simulation based on the smoothed particle hydrodynamics method (SPH) with the 3D reconstructured data. Information like flow permeability and tortuosity of the matrices can be therefore obtained from our simulation. We found that inertial effects can be quite significant at the normal pressure gradient conditions typical for espresso brewing, and can provide an explanation for the inconsistency of permeability measurements seen in the literature. Several types of coffee powder are further examined, revealing their distinct microscopic details and resulting flow features. By comparing the microCT images of pre- and post-extraction coffee matrices, it is found that a decreasing porosity profile (from the bottom-outlet to the top-inlet) always develops after extraction. This counterintuitive phenomenon can be explained using a pressure-dependent erosion model proposed in our prior work. Our results reveal not only some important hydrodynamic mechanisms of coffee extraction, but also show that microCT scan can provide useful microscopic details for coffee extraction modelling. MicroCT scan establishes the basis for a data-driven numerical framework to explore the link between coffee powder microstructure and extraction dynamics, which is the prerequisite to study the time evolution of both volatile and non-volatile organic compounds and then the flavour profile of coffee brews.

The effect of habitat transformation on a twig epiphytic orchid: Evidence from population dynamics

The tropical Andean landscape has been dramatically transformed over the last century with remaining native forest limited to small fragments within a heterogeneous matrix of crops, cattle pastures, and urban environments. We aimed to explore the impact of habitat transformation on the population dynamics in an endemic twig epiphytic orchid located within the undisturbed forest and within modified matrix habitat in two regions with contrasting landscape structures: with a dominant shade coffee matrix and a dominant grassland matrix. Over 2 years, we surveyed 4,650 individuals of the Colombian endemic orchid, Rodriguezia granadensis. We undertook four post-breeding censuses in three sites in each region in both native forest and pasture sub-sites (12 sub-sites; 48 censuses in total), and constructed demographic transition matrices (n = 36). The transition probabilities were calculated using a Bayesian approach and population grow rates were evaluated using asymptotic models and elasticities using transient dynamics. Between regions, higher population growth rate and inertia (defined as the largest or smallest long-term population density with the same initial density distribution) was seen in the shade coffee-dominated landscape. Additionally, population growth rate and damping ratio was higher in forest compared with pasture, with lower convergence time for the forest subsites. These demographic patterns reveal the contrasting levels of population resilience of this orchid in different landscape structures with the more connected shade-coffee dominated landscape permitting some healthier populations with greater population growth and survival in forest than pasture. This study highlights that twig epiphyte colonization of isolated phorophytes in pastures should not be interpreted as a sign of a healthy population but as a temporal transitory period.

Comparison of Sweet–Sour Taste Interactions between Cold Brewed Coffee and Water

Most beverages are complex matrices. Different taste compounds within these matrices interact, and thus affect the perception of the tastes. Sweetness and sourness have generally been known to suppress each other, but often such investigations have focused on aqueous solutions. Investigations into what happens when these known interactions are transferred to more complex solutions are scarce. In this study, we investigated the differences in taste interactions between an aqueous matrix and a cold-brewed coffee matrix. Two sub-studies were conducted. In one, six aqueous samples were evaluated by 152 naïve consumers; in the other six cold-brewed coffee samples were evaluated by 115 naïve consumers. In both studies participants tasted samples with no addition or with addition of either sucrose, citric acid, tartaric acid, or a mix of sucrose and either of the acids. Results showed that the sweetness of sucrose was suppressed by both citric acid and tartaric acid in both matrices. The sourness of both citric acid and tartaric acid was suppressed in the aqueous matrix, but only the sourness of tartaric was suppressed in the coffee matrix. Generally, the suppression was lower in the coffee matrix compared to the aqueous matrix. In conclusion, results from taste interaction studies conducted on aqueous matrices can to some extent, with caution, be interpolated to more complex matrices. Importantly, suppression effects might diminish with an increase in matrix complexity.

Sequential combination of solid-phase sorbents to enhance the selectivity of organosulfur compounds for flavour analysis.

Flavour analysis remains challenging due to the range of selectivity demands, from the extraction of multiclass volatile compounds to the purification of low-concentration odourants (e.g. organosulfur compounds) amidst the high food matrix noise. In this study, the varying selectivities of solid-phase extraction (SPE) were leveraged upon for both multiclass and organosulfur compound analysis, using coffee as a model matrix. Polymeric SPE (Bond Elut ENV) was screened for significant (p<0.05) parameters affecting the recovery of 37 multiclass compounds, and the most influential parameters were optimised using a Box-Behnken design (elution solvent of 67:33 dichloromethane:ethyl acetate, loading pH of 4.8, and wash solvent of water). Following this, low-concentration organosulfur compounds which were challenging to detect in the complex coffee matrix were purified by adding a sequential SPE step for selectivity. A silver-based ligand-exchange SPE step (MetaSEP IC-Ag) was optimised for organosulfur compound recovery (wash solvents of dichloromethane and 43% acetonitrile in dichloromethane, elution solvent of 90mmol/L 1,4-dithiothreitol in dichloromethane). This was found to be complementary to polymeric SPE's matrix clean-up effect (which improved average organosulfur compound recovery by 5.45 times). Finally, both multiclass and sequential organosulfur extraction techniques demonstrated good performance in terms of reproducibility (1.0-9.0%) and linearity (R2>0.995), allowing for the detection of 3-mercapto-3-methylbutyl formate (3.741±0.387ng/mL) in coffee samples. In conclusion, this study highlights the potential for SPE to address a variety of complex flavour analysis demands with the appropriate selection and combination of solid-phases.

In vitro digestion nullified the differences triggered by roasting in phenolic composition and α-glucosidase inhibitory capacity of coffee

Coffee beans were roasted to medium, dark and very dark degrees, and respective brews were in vitro digested and tested for α-glucosidase inhibition, to explore their antidiabetic potential. Phenolic acids (PA) and Maillard reaction indices (MRI) were quantified before and after digestion. Molecular docking was carried out to investigate α-glucosidase inhibition mechanisms.In vitro digested coffee inhibited α-glucosidase more effectively, compared to undigested samples, but without differences between roasting degrees. The inhibitory effect may be attributed to chlorogenic acids (CGA), which were the most abundant PA in digested coffees. In fact, molecular docking predicted a high affinity of CGA for α-glucosidase. Even though digestion nullified roasting-induced differences in α-glucosidase inhibition, CGA showed a decreasing trend upon digestion. Similarly, MRI did not differ among coffees upon digestion but decreased compared to undigested samples. Overall, the results reported in this study suggest that the presence of different compounds in coffee matrix may contribute to an antidiabetic effect.

Direct multielement determination of Cd, Pb, Fe, and Mn in ground coffee samples using energy dispersive X‐ray fluorescence spectrometry

An analytical method was proposed for the quantification of Cd, Pb, Mn, and Fe in ground coffee samples using energy dispersive X‐ray fluorescence (EDXRF) spectrometry. External calibration was possible using the coffee matrix itself as standards with samples previously analyzed by inductively coupled plasma optical emission spectrometry (ICP OES). The ground coffee samples were analyzed in the form of pellets, which were placed in the center of the EDXRF measuring cell for the determination of the analytes. The limits of detection were 62, 55, 86, and 76 μg/kg for Cd, Pb, Fe, and Mn, respectively. Good linearities were obtained for the calibration curves with coefficients of correlation (R) of 0.9959 for Cd, 0.9939 for Pb, 0.9929 for Fe, and 0.9909 for Mn. Precision, evaluated as the relative standard deviation (RSD) of 10 independent measurements of different cataloged samples with an approximate concentration of 400 μg/kg for each analyte, was between 6 and 14% (n = 10), demonstrating that the preparation of the samples in form of pellets was sufficient to perform analyses by EDXRF. The proposed method was applied in the determination of Cd, Pb, Fe, and Mn in ground coffee samples collected in Salvador, Bahia, Brazil. In a parallel comparative procedure, coffee samples were also analyzed by ICP OES after sample pretreatment by acid digestion. A paired Student's t test was carried out (confidence level of 95%, n = 5) and no significant difference was observed between the concentrations obtained by the proposed and comparative methods.

In-vitro bioaccessibility of spray dried refined kenaf (Hibiscus cannabinus) seed oil applied in coffee drink.

This study aimed to investigate the effect of a coffee beverage matrix on the oil release percentage and bioaccessibility of bioactive compounds from microencapsulated refined kenaf seed oil (MRKSO) using an in vitro gastrointestinal digestion model. Refined kenaf seed oil was spray-dried with gum arabic, β-cyclodextrin, and sodium caseinate. Oil release percentage, total phenolic content, radical scavenging activity of DPPH and ABTS, tocopherol and tocotrienol contents, as well as phytosterol content, were measured in the oil released from digested MRKSO along with the coffee matrix and compared to the digested MRKSO without coffee matrix and undigested MRKSO. Refined kenaf seed oil showed a significantly higher oxidative stability index than crude, degummed, and neutralized oil samples. About 91.2 and 94.7% of the oils were released from the digested MRKSO without and with coffee matrix, respectively. Oil released from the digested MRKSO with coffee matrix showed an increase in the total phenolic content (200.5%), DPPH (172.7%), and ABTS (68.1%) values, tocopherol and tocotrienol contents (24.6%), as well as the phytosterol content (62.0%), compared to oil released from the digested MRKSO without coffee matrix. MRKSO was successfully incorporated in the coffee drink and can use as a partial replacement for coffee creamers or supplementation in coffee drinks.

Parameters affecting the exposure to furfuryl alcohol from coffee

Recently, furfuryl alcohol (FFA) was labelled a human potential carcinogen (group 2B) by the International Agency for Research on Cancer. Its alimentary exposure is mostly from coffee since in any other foods the concentrations are significantly lower. The various storage conditions of roasted coffee, the different brewing techniques applied and the bioaccessibility after ingestion are potential parameters that might alter the exposure to FFA from coffee. An 8 weeks stability study at varying temperatures showed that FFA is stable in the ground coffee matrix. Moreover, different brewing techniques extracted different amounts of FFA and affected its final concentration. The evaluation of the relative exposure to four furans (FFA, 5-hydroxymethyl-furaldehyde, 2-furoic acid, and 5-hydroxymethyl-2-furoic acid) revealed that FFA amounts were at least 2-fold the amounts of other studied furans in the same brew. A 22-fold variation in the concentration of the four furans in brews prepared using different coffee grounds and brewing techniques could be observed. 90% of the four furans were extracted by the first 25-30% fraction of the filter brew. A significant decrease of FFA is observed after stressing with simulated gastric fluid. However, this decrease could not be reproduced when mimicking a regular coffee ingestion situation.

Agricultural matrices affect ground ant assemblage composition inside forest fragments.

The establishment of agricultural matrices generally involves deforestation, which leads to fragmentation of the remaining forest. This fragmentation can affect forest dynamics both positively and negatively. Since most animal species are affected, certain groups can be used to measure the impact of such fragmentation. This study aimed to measure the impacts of agricultural crops (matrices) on ant communities of adjacent lower montane Atlantic rainforest fragments. We sampled nine forest fragments at locations surrounded by different agricultural matrices, namely: coffee (3 replicates); sugarcane (3); and pasture (3). At each site we installed pitfall traps along a 500 m transect from the interior of the matrix to the interior of the fragment (20 pitfall traps ~25 m apart). Each transect was partitioned into four categories: interior of the matrix; edge of the matrix; edge of the fragment; and interior of the fragment. For each sample site, we measured ant species richness and ant community composition within each transect category. Ant richness and composition differed between fragments and matrices. Each sample location had a specific composition of ants, probably because of the influence of the nature and management of the agricultural matrices. Species composition in the coffee matrix had the highest similarity to its corresponding fragment. The variability in species composition within forest fragments surrounded by pasture was greatest when compared with forest fragments surrounded by sugarcane or, to a lesser extent, coffee. Functional guild composition differed between locations, but the most representative guild was ‘generalist’ both in the agricultural matrices and forest fragments. Our results are important for understanding how agricultural matrices act on ant communities, and also, how these isolated forest fragments could act as an island of biodiversity in an ‘ocean of crops’.

Biodiversity extinction thresholds are modulated by matrix type

Biodiversity extinction thresholds are abrupt declines in biological diversity that occur with habitat loss, associated with a decline in habitat connectivity. Matrix quality should influence the location of thresholds along habitat loss gradients through its effects on connectivity; however these relationships have seldom been explored empirically. Using field data from 23 independent 1254 ha landscapes in the Brazilian Atlantic Forest, we evaluated how tropical avian biodiversity responds to native forest loss within habitat patches embedded either in homogeneous pasture matrix context (with a high proportion of cattle pastures), and heterogeneous coffee matrix context (with high abundance of sun coffee plantations). We considered taxonomic, functional, and phylogenetic diversity, and tested if matrix type and choice of diversity metric influenced the location of biodiversity thresholds along the forest cover gradient. We found that matrix type postponed the abrupt loss of taxonomic diversity, from a threshold of 35% of forest cover in homogeneous pasture matrix to 19% in heterogeneous coffee matrix. Phylogenetic diversity responded similarly, with thresholds at 30 and 24% in homogeneous‐pasture and heterogeneous‐coffee matrices, respectively, but no relationship with forest cover was detected when corrected for richness correlation. Despite the absence of a threshold for functional diversity in either matrix types, a strong decline below 20% of habitat amount was detected. Finally, below 20% native habitat loss, all diversity indices demonstrated abrupt declines, indicating that even higher‐quality matrices cannot postpone diversity loss below this critical threshold. These results highlight that taxonomic diversity is a more sensitive index of biodiversity loss in fragmented landscapes, which may be used as a benchmark to prevent subsequent functional and phylogenetic losses. Furthermore, increasing matrix quality appears an efficient conservation strategy to maintain higher biodiversity levels in fragmented landscapes over a larger range of habitat loss.

Coffee: A Dietary Intervention on Type 2 Diabetes?

Coffee beverages, prepared in a multitude of ways around the world, are increasingly part of our daily lives. Although considered an unhealthy beverage for decades, coffee is increasingly the headline of medical journals in association with a reduced risk for several diseases. What if this beverage could give us pleasure, while modulating mood and lowering the risk for several diseases of the modern society, including type 2 diabetes (T2D)? Based on the most recent epidemiological and research data, long-term consumption of coffee beverages is associated with a lower risk of developing T2D in healthy individuals, probably involving multiple mechanisms, with interventions on glucose homeostasis, antioxidant activity, and inflammatory biomarkers. Several coffee constituents potentially responsible for these effects are described, as well as the factors that make their presence highly variable, with interesting effects associated with chlorogenic acids, trigonelline and norharman. Due to the high number of compounds contained in coffee, we explore the potential synergic effect within the coffee matrix. Moreover, acute coffee consumption shows different health effects from those achieved on a long-term daily consumption, and not all coffee beverages are similar. Still, despite the huge amount or work developed in the last decade, the substances and mechanisms behind these protective effects on T2D are still to be fully elucidated, being therefore soon for dietary interventions based on coffee.

Edge effects on dung beetle assemblages in an Andean mosaic of forest and coffee plantations

AbstractIn landscapes dominated by agriculture, conspicuous edges often occur between landscape elements. However, there is disagreement about the existence and intensity of edge effects, and information about species‐specific responses remains scarce. Studying such edge effects can help elucidate functional landscape connectivity and contribute to agricultural management. We, therefore, assessed whether sun‐grown coffee represents a barrier to dung beetles in an Andean agricultural landscape. We also evaluated whether the response to edge effects differs among species. We found that diversity and abundance tend to decrease from forest to sun‐grown coffee and that there are sharp increases in species turnover at the forest–coffee edge. We detected several different species‐specific responses to the forest–coffee edge, suggesting differences in the mobility of the species (or spillover) and in the degree of penetration that takes place from forest patches to sun‐grown coffee plantations. This study demonstrates that the sun‐grown coffee matrix constitutes a barrier to forest species and suggests that the forest–coffee ecotone is more complex than expected. Our results support the notion that the conservation value of native forest patches in agricultural scenarios depends on the functional connectivity of forest units in the landscape to maximize the opportunities species have to disperse through the agricultural matrix.

Anthropogenic Matrices Favor Homogenization of Tree Reproductive Functions in a Highly Fragmented Landscape.

Species homogenization or floristic differentiation are two possible consequences of the fragmentation process in plant communities. Despite the few studies, it seems clear that fragments with low forest cover inserted in anthropogenic matrices are more likely to experience floristic homogenization. However, the homogenization process has two other components, genetic and functional, which have not been investigated. The purpose of this study was to verify whether there was homogenization of tree reproductive functions in a fragmented landscape and, if found, to determine how the process was influenced by landscape composition. The study was conducted in eight fragments in southwest Brazil. The study was conducted in eight fragments in southwestern Brazil. In each fragment, all individual trees were sampled that had a diameter at breast height ≥3 cm, in ten plots (0.2 ha) and, classified within 26 reproductive functional types (RFTs). The process of functional homogenization was evaluated using additive partitioning of diversity. Additionally, the effect of landscape composition on functional diversity and on the number of individuals within each RFT was evaluated using a generalized linear mixed model. appeared to be in a process of functional homogenization (dominance of RFTs, alpha diversity lower than expected by chance and and low beta diversity). More than 50% of the RFTs and the functional diversity were affected by the landscape parameters. In general, the percentage of forest cover has a positive effect on RFTs while the percentage of coffee matrix has a negative one. The process of functional homogenization has serious consequences for biodiversity conservation because some functions may disappear that, in the long term, would threaten the fragments. This study contributes to a better understanding of how landscape changes affect the functional diversity, abundance of individuals in RFTs and the process of functional homogenization, as well as how to manage fragmented landscapes.

A new methodology to estimate the steady-state permeability of roast and ground coffee in packed beds

In an espresso-style extraction hot water (90±5°C) is driven through a coffee packed bed by a pressure gradient to extract soluble material from the coffee matrix. Permeability is a key parameter affecting extraction as it determines the flow rate through the bed and hence brewing and residence time. This may alter bed-to-cup mass transfer and therefore impact brew quality.In this work a methodology that will allow estimation of the permeability of coffee packed beds in steady-state was developed. Fitting measured flow rate – pressure drop data to Darcy’s law resulted in permeability values in the range of 10−13–10−14m2. Disagreement between the experimental and theoretical permeability, as estimated from dry measurements of particle size distribution and Kozeny–Carman equation, was found. Bed consolidation may have a larger effect on the packing structure than the mere decrease in bed bulk porosity. The Kozeny–Carman equation, corrected with a porosity-dependent tortuosity according to a power law, gave a good fit of the data.

Determination of volatile thiols in roasted coffee by derivatization and liquid chromatography–high resolution mass spectrometric analysis

A fast derivatization/extraction method followed by LC–ESI–HRMS was optimized for the determination of volatile thiols in roasted coffee powder, using ebselen as the derivatization agent. The analytical conditions were optimized on real coffee matrix, which was spiked with representative volatile thiols. The method sensitivity, precision, accuracy and selectivity were evaluated by using representative standard thiols. Estimated LOQs were between 0.02 and 14.8ng/kg for the thiols evaluated. Recoveries and intra-day and inter-day RSD values obtained in coffee matrix were in general around 40% and between 11 and 30%, respectively. The optimized and validated method was applied to real coffee samples. According to the established identification criteria, target thiols were identified and quantified in coffee powder samples. Among them, 4-mercapto-1-butanol and 2-methyl-3-tetrahydrofuranthiol were identified and quantified for the first time in roasted coffee. Moreover, an approach based on the formation of a diagnostic product ion was applied to detect non-target thiols, allowing the detection of nineteen thiol derivatives, fourteen of them were tentatively identified on the basis of their molecular formula. Some of them, such as methanethiol and 3-mercapto-3-methylbutyl acetate were known to be present in coffee volatile fraction, while the rest were not previously described in this product.

Metabolic fate of ochratoxin A as a coffee contaminant in a dynamic simulator of the human colon

Ochratoxin A (OTA) is a mycotoxin frequently encountered in coffee. The relevance of this contaminant in the colon upon digestion necessitates a study on its interaction with colon microbiota. Here, the fate of OTA during colon digestion was investigated using a dynamic simulator of the human gut. The influence of coffee as a food matrix was taken into account, as it may affect the colonic microbial ecosystem and, consequently, the fate of OTA. Biodegradation was followed by measuring OTA concentration over time, and by screening for several possible metabolites, using LC–ESI-MS and HRMS. The descending colon was found to be the main site of OTA biodegradation. Two metabolites, ochratoxin α and ochratoxin B, were identified, suggesting that biodegradation by gut microbiota is beneficial for the host, as they are considered less toxic than OTA. The extent of biodegradation was reduced in the presence of the coffee matrix, possibly due to competition for available carbon sources. Effects of OTA and the coffee matrix on the microbial ecosystem were contrasting. While OTA caused a specific, but lasting loss, of the beneficial species Lactobacillus reuteri, coffee temporarily altered the fermentation pattern towards lower ammonia and higher acetate and propionate production, likely due to its dietary fibre content.

A small mammal community in a forest fragment, vegetation corridor and coffee matrix system in the Brazilian Atlantic forest.

The objective of our work was to verify the value of the vegetation corridor in the conservation of small mammals in fragmented tropical landscapes, using a model system in the southeastern Minas Gerais. We evaluated and compared the composition and structure of small mammals in a vegetation corridor, forest fragments and a coffee matrix. A total of 15 species were recorded, and the highest species richness was observed in the vegetation corridor (13 species), followed by the forest fragments (10) and the coffee matrix (6). The absolute abundance was similar between the vegetation corridor and fragments (F = 22.94; p = 0.064), and the greatest differences occurred between the vegetation corridor and the matrix (F = 22.94; p = 0.001) and the forest fragments and the matrix (F = 22.94; p = 0.007). Six species showed significant habitat preference possibly related to the sensitivity of the species to the forest disturbance. Marmosops incanus was the species most sensitive to disturbance; Akodon montensis, Cerradomys subflavus, Gracilinanus microtarsus and Rhipidomys sp. displayed little sensitivity to disturbance, with a high relative abundance in the vegetation corridor. Calomys sp. was the species least affected by habitat disturbance, displaying a high relative abundance in the coffee matrix. Although the vegetation corridors are narrow (4 m width), our results support the hypothesis in which they work as a forest extension, share most species with the forest fragment and support species richness and abundance closer to forest fragments than to the coffee matrix. Our work highlights the importance and cost-effectiveness of these corridors to biodiversity management in the fragmented Atlantic Forest landscapes and at the regional level.

Urinary excretion of chlorogenic acids and metabolites in humans after green mate (I. paraguariensis ) consumption.

The consumption of green mate (I. paraguariensis) has been associated with health benefits, attributed mainly to its high chlorogenic acids (CGA) contents. It is known that CGA from coffee matrix are bioavailable to humans, but their bioavailability when in mate matrix has not been investigated. The aim of this study was to measure the urinary excretion of CGA and metabolites after green mate consumption. On separate days, subjects (n=7) consumed capsules of green mate (6.2μmol of CGA/Kg weight) or placebo. Urine samples were collected at baseline and for 24h after each treatment. CGA and metabolites were measured in urine by HPLC/DAD. Six CGA (3 caffeoylquinic and 3 dicaffeoylquinic acids) and 14 metabolites (caffeic, vanillic, ferulic, isoferulic, p‐coumaric, gallic, p‐hydroxybenzoic, dihydrocaffeic, syringic, sinapic, 3,4‐dihidroxyphenilacetic, 3‐(4‐hydroxyphenil)propionic, trans‐3‐hydroxycinnamic and benzoic acids) were quantified in the urine samples. The total urinary excretion after green mate (516.6μmol) consumption was higher (p&lt;0.05) than placebo (374.7μmol). On average, after green mate consumption, 55% of total hydroxycinnamic acid and quinic acid moieties’ derivatives was recovered in urine. However, the existence of phenolic acids in urine after the placebo treatment should be considered. This study indicates that CGA from green mate are absorbed and metabolized in humans.Financial support: CNPq and FAPERJ, Brazil.

Validation by collaborative trial of an isotope dilution liquid chromatographic tandem mass spectrometric method to determine the content of acrylamide in roasted coffee

A method for the determination of acrylamide in roasted coffee was subjected to method validation by collaborative trial. The aim was to extend the scope of a method already standardized for the determination of acrylamide in bakery and potato products to include roasted coffee. Modifications of the already standardized method were therefore kept to a minimum. The method was based on aqueous extraction of the roasted coffee matrix and solid-phase extraction (SPE) clean-up followed by isotope dilution high-performance liquid chromatography with tandem mass spectrometric detection (LC-MS/MS). The test portion of the sample was spiked with stable isotope-labelled acrylamide and extracted on a mechanical shaker with n-hexane and water for 1 h. The sample extract was centrifuged, the organic phase discarded, and a portion of the aqueous extract further cleaned-up by SPE on an Isolute Multimode cartridge followed by a second clean-up step on an Isolute ENV + cartridge. The volume of the acrylamide-containing fraction eluted from the second SPE column was reduced by evaporation and analysed by LC-MS/MS. Three coffee samples and one aqueous acrylamide standard solution were sent to eleven laboratories from eight European Union Member States. All samples were sent as blind duplicates. Based on the reported results, the relative standard deviations for reproducibility (RSDR) were 11.5% at a level of 160 µg kg−1, 10.1% at a level of 263 µg kg−1, and 9.6% at a level of 585 µg kg−1. The values for repeatability (RSDr) in those materials ranged from 1.0% to 3.5%. The method performance parameters satisfied internationally accepted criteria. Hence, the method would be suitable for the enforcement of regulatory limits.