Three-dimensional compaction of soft granular packings.

This paper analyzes the compaction behavior of assemblies composed of soft (elastic) spherical particles beyond the jammed state, using three-dimensional non-smooth contact dynamic simulations. The assemblies of particles are characterized using the evolution of the packing fraction, the coordination number, and the von Misses stress distribution within the particles as the confining stress increases. The packing fraction increases and tends toward a maximum value close to 1, and the mean coordination number increases as a square root of the packing fraction. As the confining stress increases, a transition is observed from a granular-like material with exponential tails of the shear stress distributions to a continuous-like material characterized by Gaussian-like distributions of the shear stresses. We develop an equation that describes the evolution of the packing fraction as a function of the applied pressure. This equation, based on the micromechanical expression of the granular stress tensor, the limit of the Hertz contact law for small deformation, and the power-law relation between the packing fraction and the coordination of the particles, provides good predictions from the jamming point up to very high densities without the need for tuning any parameters.

Compaction Model for Highly Deformable Particle Assemblies.

The compaction behavior of deformable grain assemblies beyond jamming remains bewildering, and existing models that seek to find the relationship between the confining pressure P and solid fraction ϕ end up settling for empirical strategies or fitting parameters. Using a coupled discrete-finite element method, we analyze assemblies of highly deformable frictional grains under compression. We show that the solid fraction evolves nonlinearly from the jamming point and asymptotically tends to unity. Based on the micromechanical definition of the granular stress tensor, we develop a theoretical model, free from ad hoc parameters, correctly mapping the evolution of ϕ with P. Our approach unveils the fundamental features of the compaction process arising from the joint evolution of grain connectivity and the behavior of single representative grains. This theoretical framework also allows us to deduce a bulk modulus equation showing an excellent agreement with our numerical data.

Geometric cohesion in two-dimensional systems composed of star-shaped particles.

Using a discrete element method, we investigate the phenomenon of geometric cohesion in granular systems composed of star-shaped particles with 3 to 13 arms. This was done by analyzing the stability of columns built with these particles and by studying the microstructure of these columns in terms of density and connectivity. We find that systems composed of star-shaped particles can exhibit geometric cohesion (i.e., a solidlike behavior, in the absence of adhesive forces between the grains), depending on the shape of the particles and the friction between them. This phenomenon is observed up to a given critical size of the system, from which a transition to a metastable behavior takes place. We also have evidence that geometric cohesion is closely linked to the systems' connectivity and especially to the capability of forming interlocked interactions (i.e., multicontact interactions that hinder the relative rotation of the grains). Our results contribute to the understanding of the interesting and potentially useful phenomenon of geometric cohesion. In addition, our work supplements an important set of experimental observations and sheds light on the complex behavior of real, three-dimensional, granular systems.

Bioavailability of bioactive food compounds: a challenging journey to bioefficacy.

Bioavailability is a key step in ensuring bioefficacy of bioactive food compounds or oral drugs. Bioavailability is a complex process involving several different stages: liberation, absorption, distribution, metabolism and elimination phases (LADME). Bioactive food compounds, whether derived from various plant or animal sources, need to be bioavailable in order to exert any beneficial effects. Through a better understanding of the digestive fate of bioactive food compounds we can impact the promotion of health and improvement of performance. Many varying factors affect bioavailability, such as bioaccessibility, food matrix effect, transporters, molecular structures and metabolizing enzymes. Bioefficacy may be improved through enhanced bioavailability. Therefore, several technologies have been developed to improve the bioavailability of xenobiotics, including structural modifications, nanotechnology and colloidal systems. Due to the complex nature of food bioactive compounds and also to the different mechanisms of absorption of hydrophilic and lipophilic bioactive compounds, unravelling the bioavailability of food constituents is challenging. Among the food sources discussed during this review, coffee, tea, citrus fruit and fish oil were included as sources of food bioactive compounds (e.g. (poly)phenols and polyunsaturated fatty acids (PUFAs)) since they are examples of important ingredients for the food industry. Although there are many studies reporting on bioavailability and bioefficacy of these bioactive food components, understanding their interactions, metabolism and mechanism of action still requires extensive work. This review focuses on some of the major factors affecting the bioavailability of the aforementioned bioactive food compounds.

Nonlinear effect of grain elongation on the flow rate in silo discharge.

By means of two-dimensional numerical simulations based on contact dynamics, we present a systematic analysis of the joint effects of grain shape (i.e., grain elongation) and system size on silo discharge for increasing orifice sizes D. Grains are rounded-cap rectangles whose aspect ratio are varied from 1 (disks) to 7. In order to clearly isolate the effect of grain shape, the mass of the grains is keeping constant as well as the condition of the discharge by reintroducing the exiting grains at the top of the silo. In order to quantify the possible size effects, the thickness W of the silos is varied from 7 to 70 grains diameter, while keeping the silos aspect ratio always equal to 2. We find that, as long as size effects are negligible, the flow rate Q increases as a Beverloo-like function with D, also for the most elongated grains. In contrast, the effects of grain elongation on the flow rate depend on orifice size. For small normalized orifice sizes, the flow rate is nearly independent with grain elongation. For intermediate normalized orifice sizes the flow rate first increases with grain elongation up to a maximum value that depends on the normalized size of the orifice and saturates as the grains become more elongated. For larger normalized orifice size, the flow rate is an increasing function of grains' aspect ratio. Velocity profiles and packing fraction profiles close to the orifice turn out to be self-similar for all grain shapes and for the whole range of orifice and system sizes studied. Following the methodology introduced by Janda et al. [Phys. Rev. Lett. 108, 248001 (2012)PRLTAO0031-900710.1103/PhysRevLett.108.248001], we explain the nonlinear variation of Q with grain elongation, and for all orifice sizes, from compensation mechanisms between the velocity and packing fraction measured at the center of the orifice. Finally, an equation to predict the evolution of Q as a function of the aspect ratio of the grains is deduced.

Grain size distribution does not affect the residual shear strength of granular materials: An experimental proof.

Granular materials are used in several fields and in a wide variety of processes. An important feature of these materials is the diversity of grain sizes, commonly referred to as polydispersity. When granular materials are sheared, they exhibit a predominant small elastic range. Then, the material yields, with or without a peak shear strength depending on the initial density. Finally, the material reaches a stationary state, in which it deforms at a constant shear stress, which can be linked to the residual friction angle ϕ_{r}. However, the role of polydispersity on the shear strength of granular materials is still a matter of debate. In particular, a series of investigations have proved, using numerical simulations, that ϕ_{r} is independent of polydispersity. This counterintuitive observation remains elusive to experimentalists, and especially for some technical communities that use ϕ_{r} as a design parameter (e.g., the soil mechanics community). In this Letter, we studied experimentally the effects of polydispersity on ϕ_{r}. In order to do so, we built samples of ceramic beads and then sheared these samples in a triaxial apparatus. We varied polydispersity, building monodisperse, bidisperse, and polydisperse granular samples; this allowed us to study the effects of grain size, size span, and grain size distribution on ϕ_{r}. We find that ϕ_{r} is indeed independent of polydispersity, confirming the previous findings achieved through numerical simulations. Our work fairly closes the gap of knowledge between experiments and simulations.

Compacting an assembly of soft balls far beyond the jammed state: Insights from three-dimensional imaging.

Very soft grain assemblies have unique shape-changing capabilities that allow them to be compressed far beyond the rigid jammed state by filling void spaces more effectively. However, accurately following the formation of these systems by monitoring the creation of new contacts, monitoring the changes in grain shape, and measuring grain-scale stresses is challenging. We developed an experimental method that overcomes these challenges and connects their microscale behavior to their macroscopic response. By tracking the local strain energy during compression, we reveal a transition from granular-like to continuous-like material. Mean contact geometry is shown to vary linearly with the packing fraction, which is supported by a mean field approximation. We also validate a theoretical framework which describes the compaction from a local view. Our experimental framework provides insights into the granular micromechanisms and opens perspectives for rheological analysis of highly deformable grain assemblies in various fields ranging from biology to engineering.

Experimental validation of a micromechanically based compaction law for mixtures of soft and hard grains.

In this Letter, we report on an experimental study which analyzes the compressive behavior of two-dimensional bidisperse granular assemblies made of soft (hyperelastic) and hard grains in varying proportions (κ is the portion of soft grains). By means of a recently developed uniaxial compression setup [Vu and Barés, Phys. Rev. E 100, 042907 (2019)]2470-004510.1103/PhysRevE.100.042907 and using an advanced digital image correlation method, we follow, beyond the jamming point, the evolution of the main mechanical observables, from the global scale down to the strain field inside each deformable grain. First, we validate experimentally and extend to the uniaxial case a recently proposed micromechanical compaction model linking the evolution of the applied pressure P to the packing fraction ϕ [Cantor et al., Phys. Rev. Lett. 124, 208003 (2020)]0031-900710.1103/PhysRevLett.124.208003. Second, we reveal two different linear regimes depending on whether the system is above or below a crossover strain unraveling a transition from a discrete to a continuous-like system. Third, the evolution of these linear laws is found to vary linearly with κ. These results provide a comprehensive experimental and theoretical framework that can now be extended to a more general class of polydisperse soft granular systems.

Bacteroides uniformis is a putative bacterial species associated with the degradation of the isoflavone genistein in human feces.

Inter-individual variation in isoflavone absorption depends on gut microbial degradation and affects the efficacy of these compounds. We hypothesized that inter-individual variation in fecal isoflavone disappearance coincided with variation in bacterial species. In vitro anaerobic fecal disappearance of isoflavones was measured from 33 participants by HPLC. Fecal microbial 16S rRNA variable region PCR products were obtained from 4 participants with the greatest and least genistein or glycitein degradation and were subjected to denaturing gradient gel electrophoresis. DNA bands with a homology of 90-95% to Bacteroides uniformis and Faecalibacterium prausnitzii were present in greater intensities in fecal samples showing a genistein disappearance rate constant of 1.47 ± 0.14 h(-1) compared with those with a genistein disappearance rate constant of 0.15 ± 0.03 h(-1) (P < 0.05). Human fecal bacterial species with DNA sequences 90-100% homologous to Tannerella forsythensis and 4 other species were present in greater intensities in fecal samples showing a glycitein disappearance rate constant of 0.57 ± 0.30 h(-1) compared with fecal samples with a glycitein disappearance rate constant of 0.08 ± 0.03 h(-1) (P < 0.05). In high degraders, B. uniformis may be a candidate for genistein degradation and T. forsythensis for glycitein degradation, based on fecal isoflavone degradation in the presence of these species. Bacteroides acidifaciens increased isoflavone disappearance in anaerobic human fecal incubations under nutrient-rich and -depleted conditions, suggesting this species as one responsible for the generally high degradation of isoflavones by humans. These fecal microbes are candidate biomarkers for interindividual variation in isoflavone uptake and efficacy.

Plasma pharmacokinetics of catechin metabolite 4'-O-Me-EGC in healthy humans.

BACKGROUND: Tea is an infusion of the leaves of the Camellia sinensis plant and is the most widely consumed beverage in the world after water. Green tea contains significant amounts of polyphenol catechins and represents a promising dietary component to maintain health and well-being. Epidemiological studies indicate that polyphenol intake may have potential health benefits, such as, reducing the incidence of coronary heart disease, diabetes and cancer. While bioavailability of green tea bioactives is fairly well understood, some gaps still remain to be filled, especially the identification and quantification of conjugated metabolites in plasma, such as, sulphated, glucuronidated or methylated compounds. AIM OF THE STUDY: In the present study, we aimed to quantify the appearance of green tea catechins in plasma with particular emphasis on their methylated forms. RESULTS: After feeding 400 mL of green tea, 1.25% infusion to 9 healthy subjects, we found significant amounts of EC, EGC and EGCg in plasma as expected. EGC was the most bioavailable catechin, and its methylated form (4'-O-Me-EGC) was also present in quantifiable amounts. Its kinetics followed that of its parent compound. However, the relative amount of the methylated form of EGC was lower than that of the parent compound, an important aspect which, in the literature, has been controversial so far. The quantitative results presented in our study were confirmed by co-chromatography and accurate mass analysis of the respective standards. We show that the relative abundance of 4'-O-Me-EGC is ~40% compared to the parent EGC. CONCLUSION: 4'-O-Me-EGC is an important metabolite derived from catechin metabolism. Its presence in significant amounts should not be overlooked when assessing human bioavailability of green tea.

Micromechanical description of the compaction of soft pentagon assemblies.

We analyze the isotropic compaction of assemblies composed of soft pentagons interacting through classical Coulomb friction via numerical simulations. The effect of the initial particle shape is discussed by comparing packings of pentagons with packings of soft circular particles. We characterize the evolution of the packing fraction, the elastic modulus, and the microstructure (particle rearrangement, connectivity, contact force, and particle stress distributions) as a function of the applied stresses. Both systems behave similarly: the packing fraction increases and tends asymptotically to a maximum value ϕ_{max}, where the bulk modulus diverges. At the microscopic scale we show that particle rearrangements occur even beyond the jammed state, the mean coordination increases as a square root of the packing fraction, and the force and stress distributions become more homogeneous as the packing fraction increases. Soft pentagons experience larger particle rearrangements than circular particles, and such behavior decreases proportionally to the friction. Interestingly, the friction between particles also contributes to a better homogenization of the contact force network in both systems. From the expression of the granular stress tensor we develop a model that describes the compaction behavior as a function of the applied pressure, the Young modulus, and the initial shape of the particles. This model, settled on the joint evolution of the particle connectivity and the contact stress, provides outstanding predictions from the jamming point up to very high densities.

Nondairy creamer, but not milk, delays the appearance of coffee phenolic acid equivalents in human plasma.

Chlorogenic acids (CGA) are antioxidants found in coffee. They are becoming of interest for their health-promoting effects, but bioavailability in humans is not well understood. We hypothesized that adding whole milk or sugar and nondairy creamer to instant coffee might modulate the bioavailability of coffee phenolics. Nine healthy participants were asked to randomly drink, in a crossover design, instant coffee (Coffee); instant coffee and 10% whole milk (Milk); or instant coffee, sugar, and nondairy creamer already premixed (Sugar/NDC). All 3 treatments provided the same amount of total CGA (332 mg). Blood was collected for 12 h after ingestion and plasma samples treated using a liquid-liquid extraction method that included a full enzymatic cleavage to hydrolyze all CGA and conjugates into phenolic acid equivalents. Hence, we focused our liquid chromatography-Electrospray ionization-tandem MS detection and quantification on caffeic acid (CA), ferulic acid (FA), and isoferulic acid (iFA) equivalents. Compared with a regular black instant coffee, the addition of milk did not significantly alter the area under the curve (AUC), maximum plasma concentration (C(max)), or the time needed to reach C(max) (T(max)). The C(max) of CA and iFA were significantly lower and the T(max) of FA and iFA significantly longer for the Sugar/NDC group than for the Coffee group. However, the AUC did not significantly differ. As a conclusion, adding whole milk did not alter the overall bioavailability of coffee phenolic acids, whereas sugar and nondairy creamer affected the T(max) and C(max) but not the appearance of coffee phenolics in plasma.

Plasma appearance and correlation between coffee and green tea metabolites in human subjects.

Coffee and green tea are two of the most widely consumed hot beverages in the world. Their respective bioavailability has been studied separately, but absorption of their respective bioactive phenolics has not been compared. In a randomised cross-over design, nine healthy subjects drank instant coffee and green tea. Blood samples were collected over 12 h and at 24 h to assess return to baseline. After green tea consumption, (-)-epigallocatechin (EGC) was the major catechin, appearing rapidly in the plasma; (-)-EGC gallate (EGCg) and (-)-epicatechin (EC) were also present, but (-)-EC gallate and C were not detected. Dihydroferulic acid and dihydrocaffeic acid were the major metabolites that appeared after coffee consumption with a long time needed to reach maximum plasma concentration, suggesting metabolism and absorption in the colon. Other phenolic acid equivalents (caffeic acid (CA), ferulic acid (FA) and isoferulic acid (iFA)) were detected earlier, and they peaked at lower concentrations. Summations of the plasma area under the curves (AUC) for the measured metabolites showed 1.7-fold more coffee-derived phenolic acids than green tea-derived catechins (P = 0.0014). Furthermore, we found a significant correlation between coffee metabolites based on AUC. Inter-individual differences were observed, but individuals with a high level of CA also showed a correspondingly high level of FA. However, no such correlation was observed between the tea catechins and coffee phenolic acids. Correlation between AUC and maximum plasma concentration was also significant for CA, FA and iFA and for EGCg. This implies that the mechanisms of absorption for these two classes of compounds are different, and that a high absorber of phenolic acids is not necessarily a high absorber of catechins.

Combined effects of contact friction and particle shape on strength properties and microstructure of sheared granular media.

We present a systematic numerical investigation concerning the combined effects of sliding friction and particle shape (i.e., angularity) parameters on the shear strength and microstructure of granular packings. Sliding friction at contacts varied from 0 (frictionless particles) to 0.7, and the particles were irregular polygons with an increasing number of sides, ranging from triangles to disks. We find that the effect of local friction on shear strength follows the same trend for all shapes. Strength first increases with local friction and then saturates at a shape-dependent value. In contrast, the effect of angularity varies, depending on the level of sliding friction. For low friction values (i.e., under 0.3), the strength first increases with angularity and then declines for the most angular shapes. For high friction values, strength systematically increases with angularity. At the microscale, we focus on the connectivity and texture of the contact and force networks. In general terms, increasing local friction causes these networks to be less connected and more anisotropic. In contrast, increasing particle angularity may change the network topology in different directions, directly affecting the macroscopic shear strength. These analyses and data constitute a first step toward understanding the joint effect of local variables such as friction and grain shape on the macroscopic rheology of granular systems.

Compaction of mixtures of rigid and highly deformable particles: A micromechanical model.

We analyze the isotropic compaction of mixtures composed of rigid and deformable incompressible particles by the nonsmooth contact dynamics approach. The deformable bodies are simulated using a hyperelastic neo-Hookean constitutive law by means of classical finite elements. We characterize the evolution of the packing fraction, the elastic modulus, and the connectivity as a function of the applied stresses when varying the interparticle coefficient of friction. We show first that the packing fraction increases and tends asymptotically to a maximum value ϕ_{max}, which depends on both the mixture ratio and the interparticle friction. The bulk modulus is also shown to increase with the packing fraction and to diverge as it approaches ϕ_{max}. From the micromechanical expression of the granular stress tensor, we develop a model to describe the compaction behavior as a function of the applied pressure, the Young modulus of the deformable particles, and the mixture ratio. A bulk equation is also derived from the compaction equation. This model lays on the characterization of a single deformable particle under compression together with a power-law relation between connectivity and packing fraction. This compaction model, set by well-defined physical quantities, results in outstanding predictions from the jamming point up to very high densities and allows us to give a direct prediction of ϕ_{max} as a function of both the mixture ratio and the friction coefficient.

Isoflavone glycitein diminished plasma cholesterol in female golden Syrian hamsters.

The soybean isoflavones, daidzein, genistein, and glycitein, were hypothesized to act as cholesterol-lowering components, separate from soy protein. Pure synthetic daidzein, genistein, or glycitein (0.9 mmol/kg diet) or a casein-based control diet was fed to groups of 10 female Golden Syrian hamsters for 4 weeks. Hamsters fed glycitein had significantly lower plasma total (by 15%) and non-HDL (by 24%) cholesterol compared with those fed casein (P<0.05). Daidzein and genistein's effects on these lipids did not differ from the effects of either casein or glycitein. Plasma HDL cholesterol and triglyceride concentrations were not significantly affected by dietary treatments. The percentage of urinary recovery of the ingested dose of each isoflavone was glycitein>daidzein>genistein (33.2%, 4.6%, 2.2%, respectively), with the apparent absorption of glycitein significantly greater than that of the other isoflavones. These data suggest that glycitein's greater cholesterol-lowering effect was due to its greater bioavailability, as reflected in its urinary recovery compared with that of the other isoflavones.

The roasting process does not influence the extent of conjugation of coffee chlorogenic and phenolic acids.

Understanding the bioavailability and metabolism of coffee compounds will contribute to identify the unknown biological mechanism(s) linked to their beneficial effects. The influence of the roasting process on the metabolism of coffee chlorogenic acids in humans was evaluated. In a randomized, double-blind, crossover study, 12 healthy volunteers consumed four instant coffees namely, high roasted coffee (HRC), low roasted coffee (LRC), unroasted coffee (URC), and in vitro hydrolyzed unroasted coffee (HURC). The sum of areas under the curve (AUC) ranged from 8.65-17.6 to 30.9-126 µM/h (P < 0.05) for HRC, LRC, URC, and HURC, respectively. The AUC of HRC, LRC, and URC was correlated with the initial level of phenolic acids in the coffee drinks. Despite different absorption rates, the extent of conjugation was comparable between HRC, LRC, and URC coffees but different for HURC. The most abundant circulating metabolites during the first 5 H were dihydroferulic acid (DHFA), caffeic acid-3'-O-sulfate (CA3S) and isoferulic-3'-O-glucuronide (iFA3G). DHFA and 5-4-dihydro-m-coumaric acid (mDHCoA) were the main metabolites in the period of 5-24 H. The phenolic compounds after consumption of HURC were most rapidly absorbed (Tmax 1 H) compared with the other coffees (Tmax between 9 and 11 H). Using coffees with different degrees of roasting we highlighted that in spite of different absorption rates the extent of conjugation of phenolic acids was comparable. In addition, by using a hydrolyzed unroasted coffee we demonstrated an increased absorption of phenolic acids in the small intestine. © 2016 BioFactors, 42(3):259-267, 2016.

Metabolism of glycitein (7,4'-dihydroxy-6-methoxy-isoflavone) by human gut microflora.

Gut microbial disappearance and metabolism of the soy isoflavone glycitein, 7,4'-dihydroxy-6-methoxyisoflavone, were investigated by incubating glycitein anaerobically with feces from 12 human subjects. The subjects' ages ranged from 24 to 53 years with a body mass index (BMI) of 20.9-25.8 kg/m(2) (mean BMI = 24.0 +/- 1.1 kg/m(2)). Glycitein disappearance followed an apparent first-order rate loss. Fecal glycitein disappearance rates for the subjects segregated into three different groups described as high (k = 0.67 +/- 0.14/h), moderate (k = 0.34 +/- 0.04/h), and low (k = 0.15 +/- 0.07/h) glycitein degraders (p < 0.0001). There was no dose effect on the disappearance rates for each subject from 10 to 250 microM glycitein (average k = 0.32 +/- 0.03/h, p > 0.05). Four putative glycitein metabolites, characterized by liquid chromatography-mass spectrometry (electrospray ionization using positive ionization mode), were dihydroglycitein, dihydro-6,7,4'-trihydroxyisoflavone, and 5'-O-methyl-O-desmethylangolensin. Two subjects produced a metabolite tentatively identified as 6-O-methyl-equol, and one subject produced daidzein as an additional metabolite of glycitein. These results show that glycitein is metabolized by human gut microorganisms and may follow metabolic pathways similar to other soy isoflavones.

Human gut microbial degradation of flavonoids: structure-function relationships.

The relationship between chemical structure and gut microbial degradation rates of 14 flavonoids, flavone, apigenin, chrysin, naringenin, kaempferol, genistein, daidzein, daidzin, puerarin, 7,4'-dihydroxyflavone, 6,4'-dihydroxyflavone, 5,4'-dihydroxyflavone, 5,3'-dihydroxyflavone, and 4'-hydroxyflavone, was investigated by anaerobically fermenting the flavonoids with human gut microflora (n = 11 subjects). Degradation rates for the 5,7,4'-trihydroxyl flavonoids, apigenin, genistein, naringenin, and kaempferol, were significantly faster than the other structural motifs. Puerarin was resistant to degradation by the gut microflora. Extensive degradation of flavonoids by gut microflora may result in lower overall bioavailability than those flavonoids that are slowly degraded because rapidly degrading flavonoids are less likely to be absorbed intact.

Quantification of phenolic acids and their methylates, glucuronides, sulfates and lactones metabolites in human plasma by LC-MS/MS after oral ingestion of soluble coffee.

Chlorogenic acids and derivatives like phenolic acids are potentially bioactive phenolics, which are commonly found in many foods. Once absorbed, chlorogenic and phenolic acids are highly metabolized by the intestine and the liver, producing glucuronidated and/or sulphated compounds. These metabolites were analyzed in human plasma using a validated liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method. After protein precipitation, phenolic acids and their metabolites were extracted by using ethanol and chromatographic separation was achieved by reversed-phase using an Acquity UPLC BEH C18 column combined with a gradient elution system using 1% acetic acid aqueous solution and 1% acetic acid with 100% acetonitrile. The method was able to quantify 56 different compounds including 24 phenolic acids, 4 lactones, 15 sulfates and 13 glucuronides metabolites between 5 and 1000nM in plasma for most of them, except for m-dihydrocoumaric acid, 5-ferulloylquinic-glucuronide, 4-methoxycinnamic acid, 3-phenylpropionic acid, 3-(4-methoxyphenyl)propionic acid (25 to 1000nM) and p-dihydrocoumaric acid (50-1000nM). Values of repeatability and intermediate reproducibility were below 15% of deviation in general, and maximum 20% for the lowest concentrations. The validated method was successfully applied to quantify phenolic acids and their metabolites in plasma obtained after oral ingestion of soluble coffee. In conclusion, the developed and validated method is proved to be very sensitive, accurate and precise for the quantification of these possible dietary phenols.