Metabolites Of Ibuprofen Research Articles

A low-dose prebiotic fiber supplement reduces lipopolysaccharide-binding protein concentrations in a subgroup of young, healthy adults consuming low-fiber diets.

Although the beneficial effects of fiber supplementation on overall health and the gut microbiome are well-known, it is not clear whether fiber supplementation can also alter the concentrations of lipopolysaccharide-binding protein (LBP), a marker of intestinal permeability. A secondary analysis of a previously conducted study was performed. In the randomized-order, placebo-controlled, double-blinded, cross-over study 20 healthy, young participants consuming a low-fiber diet at baseline were administered a daily dose of 12 g of prebiotic fiber compared with a placebo over a period of 4 weeks with a 4-week washout between arms. In this secondary analysis, we hypothesized that the fiber supplement would reduce LBP concentration. We further hypothesized that lecithin cholesterol acyltransferase activity, a measure of high-density lipoprotein functional capacity, would be altered. Fiber supplementation did not significantly alter LBP concentration or lecithin cholesterol acyltransferase activity in the overall cohort. However, in a subgroup of individuals with elevated baseline LBP concentrations, fiber supplementation significantly reduced LBP from 9.27 ± 3.52 to 7.02 ± 2.32 µg/mL (P = .003). Exploratory analyses found positive correlations between microbial genes involved in lipopolysaccharide synthesis and conversely negative correlations with genes involved in antibiotic synthesis and LBP. Positive correlations between LBP and multiple sulfated molecules including sulfated bile acids and perfluorooctanesulfonate, and ibuprofen metabolites were also found. These findings highlight multiple environmental and lifestyle factors such as exposure to industrial chemicals and medication intake, in addition to diet, which may influence the association between the gut microbiome and gut barrier function.

Changes in Ibuprofen Toxicity and Degradation in Response to Immobilization of Bacillus thuringiensis B1(2015b).

Ibuprofen is one of the most commonly used anti-inflammatory drugs by humans, resulting in its appearance in the environment, which can negatively affect organisms living in it. The studies undertaken have shown that the immobilized Bacillus thuringiensis B1(2015b) strain can decompose this drug at a rate of qmax = 0.36 mg/L*h, with a Ks constant of 0.95 mg/L for this process. An analysis of the effect of ibuprofen on the metabolic profile of the immobilized strain B1(2015b) showed an increase in the consumption of carbon, nitrogen, phosphorus, and sulfur compounds by this strain compared to the free strain. Studies on the toxicity of ibuprofen against the B1(2015b) strain indicated a small protective effect of the carrier, manifested by a slightly higher EC50 value = 1190 mg/L (for the free strain EC50 = 1175 mg/L). A toxicity analysis of intermedia formed during ibuprofen degradation indicated that the increase in toxicity is positively correlated with the degree of hydroxylation of ibuprofen metabolites. A toxicity analysis of the post-culture fluid obtained after ibuprofen degradation by the immobilized and free strain indicated that the products formed due to this process are completely safe.

Application of the HPLC-ELSD technique for the determination of major metabolites of ibuprofen and creatinine in human urine

The report presents robust and high throughput methods, based on liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD), for the simultaneous determination of major metabolites of ibuprofen (IBU), namely 2-hydroxyibuprofen and carboxyibuprofen (method A) as well as creatinine (Crn) (method B) in human urine. The assays primarily involve straightforward sample purification. For both methods, the chromatographic separation of the analytes is achieved within 8 min at room temperature on Poroshell 120 SB-C18 (75 × 4.6 mm, 2.7 µm) column using gradient elution. The eluents consisted of 0.1% formic acid in water and acetonitrile (method A) or water and methanol (method B) delivered at a flow rate of 1 or 0.5 mL/min, respectively. In relation to metabolites of IBU, the assay linearity was observed within 0.06–0.5 g/L in urine, while the Crn assay linearity was demonstrated within 0.5–30 mmol/L in urine. The limit of quantification for IBU metabolites was determined to be 0.06 g/L, and 0.5 mmol/L for Crn. These methods were successfully applied to urine samples delivered by ten apparently healthy donors showing that the HPLC-ELSD assays are suitable for human urine screening.

Functional evaluation of CYP2C19 and CYP3A4 gene polymorphism on ibuprofen metabolism

AimIbuprofen is the most commonly used analgesic. CYP polymorphisms are mainly responsible for the differences in drug metabolism among individuals. Variations in the ability of populations to metabolize ibuprofen can lead to drug exposure events. The aim of this study was to evaluate the effects of CYP2C19 and CYP3A4 polymorphisms on ibuprofen metabolism in a Chinese population. MethodsFirst, 31 CYP2C19 and 12 CYP3A4 microsomal enzymes were identified using an insect expression system. Then, variants were evaluated using a mature incubation system. Moreover, ibuprofen metabolite content was determined via ultra-performance liquid chromatography-tandem mass spectrometry analysis. Finally, kinetic parameters of CYP2C19 and CYP3A4 genotypes were determined via Michaelis-Menten curve fitting. ResultsMost variants exhibited significantly altered intrinsic clearance compared to the wild type. In the CYP2C19 metabolic pathway, seven variants exhibited no significant alterations in intrinsic clearance (CLint), six variants exhibited significantly high CLint (121–291%), and the remaining 15 variants exhibited substantially reduced CLint (1–71%). In the CYP3A4 metabolic pathway, CYP3A4*30 was not detected in the metabolite content due to the absence of activity, and 10 variants exhibited significantly reduced CLint. ConclusionTo the best of our knowledge, this is the first study to assess the kinetic characteristics of 31 CYP2C19 and 12 CYP3A4 genotypes on ibuprofen metabolism. However, further studies are needed on poor metabolizers as they are more susceptible to drug exposure. Our findings suggest that the kinetic characteristics in combination with artificial intelligence to predict the toxicity of ibuprofen and reduce any adverse drug reactions.

Dissemination of nonsteroidal anti-inflammatory drugs (NSAIDs) and metabolites from wastewater treatment plant to soils and agricultural crops via real-scale different agronomic practices

One of the most consumed pharmaceutical subgroups across the world is nonsteroidal anti-inflammatory drugs (NSAIDs). However, the dissemination of these compounds to the natural environments through agronomic practices is a serious global problem. The hypothesis of this study is to reveal the transition of selected NSAIDs, paracetamol (PAR), diclofenac (DCF), ibuprofen (IBU), and naproxen (NAP) together with six main metabolites, detected in raw/treated wastewater (RWW/TWW) and sewage sludge generated in an urban wastewater treatment plant (WWTP) to soils and agricultural crops (corn, barley, sunflower, and sugar beet) through two widely applied agronomic practices, irrigation with TWW and application of sewage sludge as soil amendment. In other words, the cycles of 10 NSAIDs have been evaluated by simultaneously monitoring their concentrations in RWW/TWW, sewage sludge, soils, and crops. It was determined that the parent compounds and detected metabolites were treated at quite higher removal efficiencies (93.4 – >99.9%) in the studied WWTP, while DCF was eliminated poorly (7.9–52.2%). However, although it changes seasonally for some compounds, it was determined that the concentrations of almost all investigated NSAIDs increased at the determined irrigation points in the discharge channel (DC) where agricultural irrigations were performed. Apart from that, DCF, NAP, and 2-hydroxyibuprofen (2-OH-IBU) were always detected in sewage sludge seasonally up to about 20.5, 11.3, and 3.7 ng/g, respectively. While 2-OH-IBU was determined as the dominant metabolite in RWW, TWW, and sewage sludge, the metabolite of 1-hydroxyibuprofen (1-OH-IBU) was determined as the dominant compound in soils. Although 1-OH-IBU was not detected in TWW and sewage sludge in any season, detecting this metabolite as a common compound in all investigated soils (up to 60.1 ng/kg) reveals that this compound is the primary transformation product of IBU in soils. It was observed that at least one of the metabolites of IBU (1-OH-IBU and/or 2-OH-IBU) was detected in all plants grown (up to 0.75 ng/g), especially during the periods when both agricultural practices were applied. In addition, the detection of 1-OH-IBU with increasing concentrations from root to shoots in corn grown as a result of both agronomic practices shows that this compound has a high translocation potential in the corn plant. Apart from this, it was determined that PAR was detected in corn (up to 43.3 ng/kg) and barley (up to 16.8 ng/kg) within the scope of irrigation with TWW, and NAP was detected in sugar beet (up to 11.2 ng/kg) through sewage sludge application.

The Intestinal and Biliary Metabolites of Ibuprofen in the Rat with Experimental Hyperglycemia.

Hyperglycemia is reported to be associated with oxidative stress. It can result in changes in the activities of drug-metabolizing enzymes and membrane-integrated transporters, which can modify the fate of drugs and other xenobiotics; furthermore, it can result in the formation of non-enzyme catalyzed oxidative metabolites. The present work aimed to investigate how experimental hyperglycemia affects the intestinal and biliary appearance of the oxidative and Phase II metabolites of ibuprofen in rats. In vivo studies were performed by luminal perfusion of 250 μM racemic ibuprofen solution in control and streptozotocin-treated (hyperglycemic) rats. Analysis of the collected intestinal perfusate and bile samples was performed by HPLC-UV and HPLC-MS. No oxidative metabolites could be detected in the perfusate samples. The biliary appearance of ibuprofen, 2-hydroxyibuprofen, ibuprofen glucuronide, hydroxylated ibuprofen glucuronide, and ibuprofen taurate was depressed in the hyperglycemic animals. However, no specific non-enzymatic (hydroxyl radical initiated) hydroxylation product could be detected. Instead, the depression of biliary excretion of ibuprofen and ibuprofen metabolites turned out to be the indicative marker of hyperglycemia. The observed changes impact the pharmacokinetics of drugs administered in hyperglycemic individuals.

An investigation based on removal of ibuprofen and its transformation products by a batch activated sludge process: A kinetic study

Ibuprofen metabolites can form in humans as a result of metabolic activities or can be produced by microorganisms in wastewater treatment plants and receiving environments, which increases their likelihood of being present in the environment. In this study, various experiments were conducted to determine the removal degree for ibuprofen, ibuprofen carboxylic acid (IBU-CBX), and 2-hydroxylated ibuprofen (IBU-2-OH) metabolites with an activated sludge reactor. Furthermore, the pseudo-first-order biodegradation rate constant (kbiol) (17.76 L/g SS day) was calculated to determine the decomposition degree of ibuprofen in the batch activated sludge system. The effects of different ibuprofen concentrations (8.2, 5.6, 3.2, 1.51 mg/L) at constant biomass concentration (3 g/L) on the biodegradation mechanism were investigated. In addition, IBU-2-OH and IBU-CBX were tested in a batch activated sludge reactor with a volume of 2 L individually at 100 μg/L with activated sludge containing 3 g/L biomass. It was observed that ibuprofen had a removal efficiency of more than 90%. IBU-CBX and IBU-2-OH were removed at approximately 27-91% and 18-82%, respectively. In abiotic conditions, the removal of ibuprofen was found to be 7.07%. It was confirmed that the removal of ibuprofen largely depended on biological degradation. This study enabled us to know which metabolites are involved in the biodegradation process of ibuprofen in batch experiments with the activated sludge process.

Effluent decontamination by the ibuprofen-mineralizing strain, Sphingopyxis granuli RW412: Metabolic processes.

The high global consumption of ibuprofen and its limited elimination by wastewater treatment plants (WWTPs), has led to the contamination of aquatic systems by this common analgesic and its metabolites. The potentially negative environmental and public health effects of this emerging contaminant have raised concerns, driving the demand for treatment technologies. The implementation of bacteria which mineralize organic contaminants in biopurification systems used to decontaminate water or directly in processes in WWTPs, is a cheap and sustainable means for complete elimination before release into the environment. In this work, an ibuprofen-mineralizing bacterial strain isolated from sediments of the River Elbe was characterized and assayed to remediate different ibuprofen-polluted media. Strain RW412, which was identified as Sphingopyxis granuli, has a 4.48Mb genome which includes plasmid sequences which harbor the ipf genes that encode the first steps of ibuprofen mineralization. Here, we confirm that these genes encode enzymes which initiate CoA ligation to ibuprofen, followed by aromatic ring activation by a dioxygenase and retroaldol cleavage to unequivocally produce 4-isobutylcatechol and propionyl-CoA which then undergo further degradation. In liquid mineral salts medium, the strain eliminated more than 2mM ibuprofen within 74h with a generation time of 16h. Upon inoculation into biopurification systems, it eliminated repeated doses of ibuprofen within a few days. Furthermore, in these systems the presence of RW412 avoided the accumulation of ibuprofen metabolites. In ibuprofen-spiked effluent from a municipal WWTP, ibuprofen removal by this strain was 7 times faster than by the indigenous microbiota. These results suggest that this strain can persist and remain active under environmentally relevant conditions, and may be a useful innovation to eliminate this emerging contaminant from urban wastewater treatment systems.

Employ of arbuscular mycorrhizal fungi for pharmaceuticals ibuprofen and diclofenac removal in mesocosm-scale constructed wetlands

This study investigated the effects of arbuscular mycorrhizal fungi (AMF) colonization on the growth of wetland plants (Glyceria maxima), and treatment performance in constructed wetlands (CWs) under the stress of pharmaceuticals ibuprofen (IBU) and diclofenac (DCF). Results showed that the growth of G. maxima was significantly increased by AMF colonization. AMF significantly increased the activities of antioxidant enzymes (peroxidase and superoxide dismutase) and soluble protein content in wetland plants, but the contents of malondialdehyde and O2•− were reduced. The removal efficiencies of TOC, PO43−-P, NH4+-N, and TN were increased in AMF+ treatments by 6%, 11%, 15% and 11%, respectively. AMF increased the removal efficiencies of IBU and DCF by 6–14% and 2–21%, respectively, and reduced the content of their metabolites (2-OH IBU, CA IBU and 4′-OH DCF) in the effluent. Besides, the presence of AMF increased the contents of IBU and DCF in plant roots, while decreased their transportation to shoots. AMF symbiosis decreased the contents of IBU metabolites (2-OH IBU and CA IBU) but increased the contents of DCF metabolite (4′-OH DCF) in the roots of the host plant. In conclusion, these results indicated that AMF plays a promising role in CWs for emerging pollutants removal.

Simultaneous and individual adsorption of ibuprofen metabolites by a modified montmorillonite

In this work, the applicability of a modified montmorillonite (C18Mt) to remove ibuprofen (IBU) and its main metabolites, 1-hydroxyibuprofen (1-OH IBU), 2-hydroxyibuprofen (2-OH IBU) and carboxyibuprofen (CBX-IBU), simultaneously or separately, from aqueous samples has been evaluated. First, the material was characterized by X-ray diffraction, Fourier-transform infrared spectroscopy and zeta potential measurements. And then, the effect of the experimental conditions (initial concentration, time, temperature and pH) on the adsorption capacity was assessed. A statistical analysis of the adsorption isotherms reveals that Langmuir was the best model in fitting the experimental adsorption data. Maximum adsorption capacities were estimated to be 64, 20, 63 and 19 mg/g for IBU, 1-OH IBU, 2-OH IBU and CBX-IBU, respectively. These values were significantly lower for the mixture solution due to the competition effect for the active sites. Moreover, it was observed that the removal time is quite fast ranging from 20 to 30 min and the adsorption kinetics was predominantly based on the pseudo-second-order model. Characterization studies showed that the main driving force of adsorption was a combination of electrostatic interaction and partitioning. It was also observed that pH had very important effects on the adsorption. C18Mt had proven to be an excellent adsorbent capable to remove new generation pollutants, even at low adsorbent dose making their application economic.

Mechanisms of bacterial conversion and degradation of pharma pollutants from nonsteroidal anti-inflammatory drugs

Recently, there has been a steady increase in fundamental interest in studying the degree of bioavailability and toxic effects of pharmaceutical pollutants on natural microorganisms, which play the role of a primary response system to the xenobiotic load in the environment. Employing the bioresources of the Regional Specialised Collection of Alkanotrophic Microorganisms (IEGM, Large-Scale Research Facilities number 73559, WDCM # 768, http://www.iegmcol.ru), the ability of actinobacteria of the genus Rhodococcus to decompose complex aromatic compounds that constitute the non-steroidal anti-inflammatory drugs (NSAIDs) widely used for medical purposes was first established. Active biodegraders of diclofenac and ibuprofen, which dominate among NSAIDs detected in the natural environment and pose the greatest potential risk to hydrobiota and humans, were selected. The kinetics and main characteristics of the biodegradation process of the tested pharmaceutical pollutants were investigated depending on the physiological state and culture conditions of their biodegraders. Cytochrome P450-dependent monooxygenases were shown to participate in the initial oxidation of the pharma pollutants. The products of bacterial decomposition of the ecotoxicants were identified, the pathways of their biodegradation were characterized, and the mathematical models of the process of complete bacterial degradation of pharmaceutical pollutants in high concentrations were described. For the first time, evidence for the C-N bond breaking and aromatic ring opening in the diclofenac structure accompanied by the formation of metabolites harmless to living organisms was obtained. Potential bioactivity of individual metabolites of diclofenac and ibuprofen was evaluated. The most typical reactions of rhodococci exposed to NSAIDs were as follows: changes in the zeta potential, morphometric parameters and degree of hydrophobicity of bacterial cells, an increased content of total cellular lipids, and the formation of bacterial associates. The obtained results are considered to be mechanisms of rhodococci adaptation and their increased resistance to toxic effects of the pharmaceutical pollutants tested. The obtained fundamental data elucidate the environmental role of rhodococci in detoxification of pharma pollutants and underpin the implementation of innovative technical solutions for advanced pharmaceutical sewage treatment and the pharmaceutical waste disposal.

Simultaneous pressurized liquid extraction and clean-up for the determination of metabolites in complex environmental solid matrices

Parabens and pharmaceuticals are two of the most significant groups of pollutants of emerging concern. Nevertheless, in spite that their presence in the environment is well-known, to the date, there is scarce information about the presence of their metabolites that can be present at higher concentrations in the environment, be more toxic and be transformed into the parent compound. In this work, an analytical method has been developed and validated for the first-time simultaneous determination of metabolites of methyl- and propylparabens, caffeine, carbamazepine, diclofenac, ibuprofen and sulfamethoxazole and their parent compounds in soil, compost and digested sludge. Box-Behnken experimental design was applied for method optimisation due to the high number of experimental variables to optimize and the different physical-chemical properties of the target compounds. The method involves simultaneous sample extraction and clean-up by selective pressurized liquid extraction and analytical determination in a single run by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Good linearity, accuracy (mainly from 80 to 112%) and precision (<11%) was obtained. Method quantification limits were in the range from 0.12 to 14.4 ng g−1 dry matter. The application of the method to soil, compost and digested sludge revealed that metabolites of parabens, carbamazepine, diclofenac and ibuprofen were at similar or higher concentrations than their parent compounds in digested sludge and the presence of metabolites of parabens, caffeine and carbamazepine in compost.

Effectiveness of single-stage and sequential sludge digestion on removal of recalcitrant pharmaceuticals and conventional pollutants

The fate of pharmaceuticals, azithromycin (AZI), carbamazepine (CBZ), diclofenac (DCF), mefenamic acid (MFA), ibuprofen (IBP) and one ibuprofen metabolite (1-hydroxy ibuprofen (1-OH IBP)), were investigated in single-stage anaerobic (AN) and sequential anaerobic/aerobic/anoxic (AN/AERO/ANOX) sludge digestion under thermophilic conditions (55 °C). Intermittent aeration was used for the sequential AERO/ANOX vessel. Digesters showed an increase in CBZ, MFA and DCF at a hydraulic retention time (HRT) of 18 days as organic matter is degraded. However, the rate of increase for MFA was 10 times lower in sequential digestion compared to single-stage AN digestion (control). Sequential AN/AERO/ANOX digestion showed a 50% decrease in IBP levels while increasing IBPs aerobic metabolite 1-OH IBP by 12%. The control AN digester showed a 62% increase in IBP and a 34% decrease in 1-OH IBP. Sequential digestion provided other benefits, including reduced ammonia generation (44.5%), improved solids removal (4.1%) and enhanced digestate dewaterability (20%) compared to the control.

22 CANONICAL CORRELATION ANALYSIS OF ADHD POLYGENIC RISK AND FUNCTIONAL BRAIN CONNECTIVITY

A novel assay for the simultaneous determination of ibuprofen (IBU) and its four probable metabolites, 1-hydroxyibuprofen (1-OH IBU), 2-hydroxyibuprofen (2-OH IBU), 3-hydroxyibuprofen (3-OH IBU) and carboxyibuprofen (CBX IBU) in equine urine samples with the application of Gas Chromatography-Electron Ionization-Mass Spectrometry (GC-EI-MS) has been developed and elaborated. The new approach for sample preparation including minimizing matrix effects by the application of weak cation exchange solid-phase extraction together with strong cation exchange solid-phase extraction has been applied. The GC-EI-MS method was validated to demonstrate specificity, matrix effect, linearity, limit of detection (LOD) and quantification (LOQ), precision, trueness, carry-over and stability by using the matrix-matched quality control samples. Additionally, extraction yield was evaluated. The assay achieved the LOQ of 1.75 μg mL−1, 0.62 μg mL−1, 4.15 μg mL−1, 0.58 μg mL−1 and 4.04 μg mL−1 for IBU, 1-OH IBU, 2-OH IBU, 3-OH IBU and CBX IBU, respectively. The developed method has been successfully applied to the excretion study in horses, in which a single oral IBU dose was administered to twelve horses (mares and geldings) and equine urine samples were collected for 5 or 6 days after the drug administration. Data on the detection and determination of three IBU metabolites, 2-OH IBU, 3-OH IBU and CBX IBU in equine urine samples has been presented for the first time. The obtained results indicated the rapid excretion of IBU and its metabolites that were detectable only in the first day after the drug administration. IBU was mainly the most abundant compound detected in equine urine samples (with two exceptions in the case of samples collected from two horses, for which the highest instrumental responses were obtained for CBX IBU). The received results have indicated that two major IBU metabolites, CBX IBU and 2-OH IBU can be important markers for the IBU abuse in view of doping control in equestrian sports.

Quantification of unconjugated and total ibuprofen and its metabolites in equine urine samples by gas chromatography–tandem mass spectrometry: Application to the excretion study

Ibuprofen (IBU) is a widely used nonsteroidal anti-inflammatory drug (NSAID) in human and also veterinary medicine. NSAIDs are usually highly metabolized compounds in horses; they are often present in equine urine, mostly in their conjugated forms (glucuronides). Thus, hydrolysis to cleave the glucuronide linkage prior to anti-doping analysis is often necessary for improving detection. In this study, unconjugated and total IBU and its metabolites were determined by GC–MS/MS in equine urine samples collected after drug administration to six horses (mares and geldings). The results indicated that enzymatic hydrolysis using β-glucuronidase from E. coli improved the detection of IBU and its phase I metabolites in equine urine samples; in a few cases, this method allowed to monitor analytes for a longer period of time. The percentages of conjugated analytes were within 5.4–48.4% for IBU and in the range of 0.2–55.5% for phase I metabolites of IBU. Unchanged IBU was found at the highest concentration levels in equine urine samples collected after drug administration; and also parent compound lasted for a longer period of time in equine urine than its phase I metabolites, with the exception of samples collected from one horse, in which 2-OH IBU was detectable for a longer duration than IBU.

In Vivo Metabolism of Ibuprofen in Growing Conventional Pigs: A Pharmacokinetic Approach.

The juvenile conventional pig has been suggested as a preclinical animal model to evaluate pharmacokinetic (PK), pharmacodynamic (PD), and safety parameters in children. However, a lot of developmental changes in pig physiology still need to be unraveled. While the in vitro ontogeny of pig biotransformation enzymes is getting more attention in literature, the in vivo developmental changes have not yet been investigated. Therefore, the aim of the current study was to evaluate the biotransformation of ibuprofen (IBU) in conventional pigs aged 1 week, 4 weeks, 8 weeks, and 6–7 months after a single intravenous and oral administration of 5 mg/kg body weight (BW) of IBU, using a PK approach in a crossover design for each age group. An ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) method was developed and validated to determine 2-hydroxyibuprofen (2OH-IBU), carboxyibuprofen (COOH-IBU), and ibuprofen glucuronide (IBU-GlcA) in pig plasma. All three metabolites could be quantified in plasma and the following PK parameters were determined: C max, T max, AUC0→6h, area under plasma concentration–time curve (AUC) ratio between parent drug and metabolite, and the absolute oral bioavailability of the parent drug IBU. The plasma concentrations of the metabolites were always lower than those of IBU. The bioavailability was high, indicating limited pre-systemic biotransformation. The AUC ratio of 2OH-IBU and COOH-IBU/IBU showed a significant increase at 4 weeks of age compared to the 1-week-old and 6- to 7-month-old pigs. Interestingly, the IBU-GlcA/IBU AUC ratio did not change with age. The present study demonstrated that the main metabolites of IBU in human are also present in growing pigs. The oxidative phase I metabolism of IBU in growing conventional pigs did change with age. In contrast, age did not seem to affect the glucuronidation capacity of IBU in conventional pigs, although more studies with other substrate drugs are needed to confirm this.

The acyl-glucuronide metabolite of ibuprofen has analgesic and anti-inflammatory effects via the TRPA1 channel

Ibuprofen is a widely used non-steroidal anti-inflammatory drug (NSAID) that exerts analgesic and anti-inflammatory actions. The transient receptor potential ankyrin 1 (TRPA1) channel, expressed primarily in nociceptors, mediates the action of proalgesic and inflammatory agents. Ibuprofen metabolism yields the reactive compound, ibuprofen-acyl glucuronide, which, like other TRPA1 ligands, covalently interacts with macromolecules. To explore whether ibuprofen-acyl glucuronide contributes to the ibuprofen analgesic and anti-inflammatory actions by targeting TRPA1, we used in vitro tools (TRPA1-expressing human and rodent cells) and in vivo mouse models of inflammatory pain. Ibuprofen-acyl glucuronide, but not ibuprofen, inhibited calcium responses evoked by reactive TRPA1 agonists, including allyl isothiocyanate (AITC), in cells expressing the recombinant and native human channel and in cultured rat primary sensory neurons. Responses by the non-reactive agonist, menthol, in a mutant human TRPA1 lacking key cysteine-lysine residues, were not affected. In addition, molecular modeling studies evaluating the covalent interaction of ibuprofen-acyl glucuronide with TRPA1 suggested the key cysteine residue C621 as a probable alkylation site for the ligand. Local administration of ibuprofen-acyl glucuronide, but not ibuprofen, in the mouse hind paw attenuated nociception by AITC and other TRPA1 agonists and the early nociceptive response (phase I) to formalin. Systemic ibuprofen-acyl glucuronide and ibuprofen, but not indomethacin, reduced phase I of the formalin response. Carrageenan-evoked allodynia in mice was reduced by local ibuprofen-acyl glucuronide, but not by ibuprofen, whereas both drugs attenuated PGE2 levels. Ibuprofen-acyl glucuronide, but not ibuprofen, inhibited the release of IL-8 evoked by AITC from cultured bronchial epithelial cells. The reactive ibuprofen metabolite selectively antagonizes TRPA1, suggesting that this novel action of ibuprofen-acyl glucuronide might contribute to the analgesic and anti-inflammatory activities of the parent drug.

Occurrence and removal of ibuprofen and its metabolites in full-scale constructed wetlands treating municipal wastewater

Ibuprofen (IBU) and its major metabolites, hydroxyibuprofen (OH-IBU) and carboxyibuprofen (CA-IBU) were monitored from November 2015 to March 2017 at four full-scale constructed wetlands with horizontal subsurface flow treating municipal wastewater in the Czech Republic. During the seven sampling campaigns, all three compounds were identified in high concentrations in the inflowing raw wastewater. The average inflow concentrations of IBU, OH-IBU and CA-IBU amounted to 14.6, 18.3 and 36.4 µg l−1. The ratio between IBU, OH-IBU and CA-IBU is in agreement with the previously published results indicating that the ibuprofen metabolites occur in higher concentration in the raw wastewater than the parent compound with CA-IBU exhibiting the highest concentrations. The outflow concentrations off studied compounds decreased and removal amounted to 44.7%, 29.3% and 47.5% for IBU, OH-IBU and CA-IBU, respectively. The results are in line with general fact that ibuprofen and its metabolites are not removed with high efficiency in anoxic or anaerobic conditions and that OH-IBU is the least removed compound. The areal loadings of IBU, OH-IBU and CA-IBU averaged 0.425, 0.574 and 0.999 mg m−2 d−1, respectively. The annual loads of all forms of ibuprofen varied between 458 and 958 g year−1 among four studies constructed wetlands.

Simultaneous determination of ibuprofen and its metabolites in complex equine urine matrices by GC-EI-MS in excretion study in view of doping control

A novel assay for the simultaneous determination of ibuprofen (IBU) and its four probable metabolites, 1-hydroxyibuprofen (1-OH IBU), 2-hydroxyibuprofen (2-OH IBU), 3-hydroxyibuprofen (3-OH IBU) and carboxyibuprofen (CBX IBU) in equine urine samples with the application of Gas Chromatography-Electron Ionization-Mass Spectrometry (GC-EI-MS) has been developed and elaborated. The new approach for sample preparation including minimizing matrix effects by the application of weak cation exchange solid-phase extraction together with strong cation exchange solid-phase extraction has been applied. The GC-EI-MS method was validated to demonstrate specificity, matrix effect, linearity, limit of detection (LOD) and quantification (LOQ), precision, trueness, carry-over and stability by using the matrix-matched quality control samples. Additionally, extraction yield was evaluated. The assay achieved the LOQ of 1.75 μg mL−1, 0.62 μg mL−1, 4.15 μg mL−1, 0.58 μg mL−1 and 4.04 μg mL−1 for IBU, 1-OH IBU, 2-OH IBU, 3-OH IBU and CBX IBU, respectively. The developed method has been successfully applied to the excretion study in horses, in which a single oral IBU dose was administered to twelve horses (mares and geldings) and equine urine samples were collected for 5 or 6 days after the drug administration. Data on the detection and determination of three IBU metabolites, 2-OH IBU, 3-OH IBU and CBX IBU in equine urine samples has been presented for the first time. The obtained results indicated the rapid excretion of IBU and its metabolites that were detectable only in the first day after the drug administration. IBU was mainly the most abundant compound detected in equine urine samples (with two exceptions in the case of samples collected from two horses, for which the highest instrumental responses were obtained for CBX IBU). The received results have indicated that two major IBU metabolites, CBX IBU and 2-OH IBU can be important markers for the IBU abuse in view of doping control in equestrian sports.

Analysis of ibuprofen and its main metabolites in roots, shoots, and seeds of cowpea (Vigna unguiculata L. Walp) using liquid chromatography-quadrupole time-of-flight mass spectrometry: uptake, metabolism, and translocation.

A liquid chromatography quadruple time-of-flight mass spectrometry (LC-QqTOF-MS/MS) method was developed for simultaneous quantitative analysis of ibuprofen (IBU), 1- and 2-hydroxyibuprofen (1-OH IBU and 2-OH IBU), and carboxyibuprofen (CBX IBU) while preserving the ability of the instrument to get precursor and product ion mass spectra of non-target compounds. The trigger was the precursor ions reaching 100cps intensity. Sample preparation was carried out by ultrasound solid-liquid extraction with methanol as extraction solvent at pH < 2 followed by solid-phase extraction (SPE) clean-up using STRATA-X cartridges and methanol as an eluent. Linearity was obtained in the range 50-10,000ngmL-1 for IBU, each OH IBU and CBX IBU (r ≥ 0.99). The proposed method was satisfactorily validated showing absolute recoveries of > 70% for all target analytes at low and high concentration levels. The lowest limit of quantification was < 50ngg-1 in plant. This method was applied to investigate IBP behavior in cowpea (Vigna unguiculata (L.) Walp) treated at high IBU concentrations and its presence in vegetables irrigated with treated water. Up to 46 metabolites, mostly hydroxylated metabolites and conjugates with hexosides and amino acids, were identified. The most abundant metabolites were also identified in an eggplant sample. Graphical Abstract ᅟ Ibuprofen metabolite identification.