citrinin and dihydrocitrinone

Until now, the available data regarding citrinin (CIT) levels in food and the consumption of contaminated foods are insufficient to allow a reliable estimate of intake. Therefore, biomonitoring configuring analysis of parent compound and/or metabolites in biological fluids, such as urine or blood, is being increasingly applied in the assessment of human exposure to CIT and its metabolite, dihydrocitrinone (DH-CIT). Most studies report urinary levels lower for the parent compound when compared with DH-CIT. A high variability either in the mean levels or in the inter-individual ratios of CIT/DH-CIT between the reported studies has been found. Levels of DH-CIT in urine were reported as being comprised between three to seventeen times higher than the parent mycotoxin. In order to comply with this objective, sensitive analytical methodologies for determining biomarkers of exposure are required. Recent development of powerful analytical techniques, namely liquid chromatography coupled to mass spectrometry (LC-MS/MS) and ultra-high-performance liquid chromatography (UHPLC-MS/MS) have facilitated biomonitoring studies, mainly in urine samples. In the present work, evidence on human exposure to CIT through its occurrence and its metabolite, in biological fluids, urine and blood/plasma, in different countries, is reviewed. The analytical methodologies usually employed to evaluate trace quantities of these two molecules, are also presented. In this sense, relevant data on sampling (size and pre-treatment), extraction, cleanup and detection and quantification techniques and respective chromatographic conditions, as well as the analytical performance, are evidenced.

Topics: Chemistry, Clinical; Chromatography, Liquid; Citrinin; Dietary Exposure; Food Contamination; Humans; Limit of Detection; Tandem Mass Spectrometry

Recent studies have implied that environmental toxins, such as mycotoxins, are risk factors for neurodegenerative diseases. To act directly as neurotoxins, mycotoxins need to penetrate or affect the integrity of the blood-brain barrier, which protects the mammalian brain from potentially harmful substances. As common food and feed contaminants of fungal origin, the interest in the potential neurotoxicity of ochratoxin A, citrinin and their metabolites has recently increased. Primary porcine brain capillary endothelial cells were used to investigate cytotoxic or barrier-weakening effects of ochratoxin A, ochratoxin α, citrinin and dihydrocitrinone. The transfer and transport properties of the mycotoxins across the barrier formed by porcine brain capillary endothelial cell monolayers were analysed using HPLC-MS/MS. High levels of Ochratoxin A caused cytotoxic and barrier-weakening effects, whereas ochratoxin α, citrinin and dihydrocitrinone showed no adverse effects up to 10 µM. Likely due to efflux transporter proteins, the transfer to the brain compartment was much slower than expected from their high lipophilicity. Due to their slow transfer across the blood-brain barrier, cerebral exposure of ochratoxin A, ochratoxin α, citrinin and dihydrocitrinone is low and neurotoxicity is likely to play a subordinate role in their toxicity at common physiological concentrations.

Topics: Animals; Blood-Brain Barrier; Brain; Citrinin; Ochratoxins; Swine

Topics: Animals; Cattle; Circular Dichroism; Citrinin; Humans; Molecular Dynamics Simulation; Mycotoxins; Poisons; Protein Binding; Rats; Serum Albumin; Spectrometry, Fluorescence; Swine

Citrinin (CIT) is a nephrotoxic mycotoxin produced by

Topics: Algorithms; Citrinin; Cyclodextrins; Models, Theoretical; Molecular Structure; Spectrometry, Fluorescence; Thermodynamics

The analysis of the nephrotoxic mycotoxin citrinin in food, feed, and physiological samples is still challenging. Nowadays, liquid chromatography coupled with mass spectrometry is the method of choice for achieving low limits of detection. But matrix effects can present impairments for this method. Stable isotope dilution analysis can prevent some of these problems. Therefore, a stable isotopically labeled standard of citrinin for use in stable isotope dilution analysis was synthesized on large scale. The improved diastereoselective total synthetic strategy offered the possibility to introduce three

Topics: Carbon Isotopes; Chromatography, High Pressure Liquid; Citrinin; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Structure; Mycotoxins; Staining and Labeling; Tandem Mass Spectrometry

A direct, fast and sensitive LC-MS/MS method was developed to measure biomarkers for mycotoxin exposure in human urine. In total, 32 biomarkers were quantitatively or semi-quantitatively measured in 32 urine samples of Belgian volunteers using two injections. All urine samples contained deoxynivalenol-15-glucuronide, the major detoxification metabolite of deoxynivalenol, in the ng/mL range. Also deoxynivalenol-3-glucuronide and de-epoxy-deoxynivalenol-glucuronide were present in, respectively, 90 and 25% of the samples, while deoxynivalenol was detected in 60% of the samples, in lower concentrations. Deoxynivalenol glucuronides were the major biomarkers for deoxynivalenol exposure. Ochratoxin A was detected in 70% of the samples in pg/mL. Citrinin and/or dihydrocitrinone were detected in 90% of the samples, also in concentrations of pg/mL. The presence of ochratoxin A and citrinin was confirmed by a second method using sample cleanup by immunoaffinity columns, followed by LC-MS/MS. Our data show that humans are much more exposed to citrinin than realized before and suggest further work on citrinin exposure in relation with ochratoxin A exposure, as both mycotoxins are nephrotoxic.

Topics: Adult; Belgium; Biomarkers; Chromatography, Liquid; Citrinin; Female; Glucuronides; Humans; Male; Mycotoxins; Ochratoxins; Tandem Mass Spectrometry; Trichothecenes

As data on food contamination with the mycotoxin citrinin (CIT) are scarce, a recently developed method for biomarker analysis (Blaszkewicz et al. in Arch Toxicol 87:1087-1094, 2013) was applied to investigate CIT exposure of German adults. CIT and its human metabolite dihydrocitrinone (HO-CIT) were determined in urine samples from a group of 50 healthy adults (n = 27 females and n = 23 males). After cleanup by immunoaffinity (CitriTest®) columns, extracts were analyzed by LC-MS/MS. The mycotoxin and its major metabolite HO-CIT were detected in 82 and 84 % of all urine samples, at concentrations ranging from 0.02 (limit of detection, LOD) to 0.08 ng/mL for CIT, and 0.05 (LOD) to 0.51 ng/mL for HO-CIT. Median urine analyte levels in the cohort were 0.03 (CIT) and 0.06 ng/mL (OH-CIT) or adjusted to creatinine 20.2 ng/g crea (CIT) and 60.9 ng/g crea (HO-CIT), respectively. Except for higher urinary CIT levels in males, differences between subgroups were not significant. This first biomarker analysis indicates widespread and variable exposure to CIT in German adults, and conversion of ingested mycotoxin to its less toxic metabolite HO-CIT, which may serve as biomarker of exposure in addition to the parent compound.

Topics: Adult; Biomarkers; Chromatography, Liquid; Citrinin; Creatinine; Data Interpretation, Statistical; Edible Grain; Female; Germany; Humans; Limit of Detection; Male; Reproducibility of Results; Sex Factors; Tandem Mass Spectrometry

Citrinin (CIT) is a mycotoxin contaminant in food commodities and can co-occur with ochratoxin A (OTA), another nephrotoxic contaminant in food and feed. Presence of OTA in maize from Bangladesh has been reported, but no data exist on CIT occurrence in food or feed in Bangladesh. Since biomonitoring provides the best approach to assess human exposure to contaminants from various sources and by all routes, a validated method for biomarker analysis has been used to investigate the presence of CIT and its metabolite dihydrocitrinone (HO-CIT) in urines from two Bangladeshi cohorts: Both analytes were determined in urine samples collected from inhabitants of a rural (n=32) and an urban (n=37) area in the Rajshahi district of Bangladesh. After cleanup by immunoaffinity columns, extracts were analyzed by LC-MS/MS; the limits of detection for CIT and HO-CIT in urine were 0.02 and 0.05 ng/mL, respectively. CIT and HO-CIT were detectable in 94 and 71% of all urine samples. Urinary biomarker levels did not show significant correlations with age, gender, and body mass index of the donors. However, excretion of CIT together with its metabolite HO-CIT was significantly higher (p<0.01) in the rural cohort (mean 1.1±1.9 ng/mL) than in the urban cohort (mean 0.14±0.14 ng/mL). This clearly indicates differences in mycotoxin exposure. As food habits differ between rural and urban people and also their main areas of occupation, further research is needed with regard to the major contributors of CIT exposure in the two cohorts. In conclusion, this first biomarker analysis indicates widespread and variable exposure to CIT in Bangladeshi adults.

Topics: Adult; Bangladesh; Biomarkers; Chromatography, Liquid; Citrinin; Cohort Studies; Female; Humans; Male; Middle Aged; Rural Population; Tandem Mass Spectrometry; Urban Population; Urine; Young Adult

Citrinin (CIT) and ochratoxin A (OTA) are mycotoxins produced by several species of the genera Aspergillus, Penicillium and Monascus. Both can be present as contaminants in various food commodities and in animal feed. The occurrence and toxicity of OTA and human exposure have been intensively studied, but for CIT such data are scarce by comparison. Recently, dihydrocitrinone (DH-CIT) was detected as main metabolite of CIT in human urine, and co-occurrence of CIT and OTA was shown in human blood plasma (Blaszkewicz et al. in Arch Toxicol 87:1087-1094, 2013). In light of these new findings, we have now investigated the toxicity of the metabolite DH-CIT in comparison with CIT and analysed the effects of mixtures of CIT and OTA in vitro. The cytotoxic potency of DH-CIT (IC50 of 320/200 μM) was distinctly lower compared with CIT (IC50 of 70/62 μM) after treatment of V79 cells for 24 and 48 h. Whereas CIT induced a concentration-dependent increase in micronucleus frequencies at concentrations ≥30 μM, DH-CIT showed no genotoxic effect up to 300 μM. Thus, conversion of CIT to DH-CIT in humans can be regarded as a detoxification step. Mixtures of CIT and OTA exerted additive effects in cytotoxicity assays. The effect of CIT and OTA mixtures on induction of micronuclei varied dependent on the used concentrations between additive for low μM concentrations and more-than-additive for high μM concentrations. Effects on cell cycle were mostly triggered by OTA when both mycotoxins were used in combination. The implications of our and related in vitro studies are discussed with respect to in vivo concentrations of CIT and OTA, which are found in animals and in humans.

Topics: Animals; Cell Cycle; Cell Line; Citrinin; Complex Mixtures; Cricetinae; Dose-Response Relationship, Drug; Inactivation, Metabolic; Micronucleus Tests; Ochratoxins

Citrinin (CIT), produced by several Penicillium, Aspergillus, and Monascus species, has been detected as contaminant in feeds, grains, and other food commodities. CIT can co-occur with ochratoxin A (OTA), a mycotoxin also known for its nephrotoxicity, and this raises concern regarding possible combined effects. But, in contrast to OTA, data on CIT contamination in foods for human consumption are scarce, and CIT biomonitoring has not been conducted so far due a lack of suitable methods for human specimen. Thus, it was the aim of the present study to develop sensitive methods for the analysis of CIT in human blood and urine to investigate human exposure. To this end, we assessed different methods of sample preparation and instrumental analysis for these matrices. Clean-up of blood plasma by protein precipitation followed by LC-MS/MS-based analysis allowed robust detection of CIT (LOD 0.07 ng/mL, LOQ 0.15 ng/mL). For urine, sample clean-up by an immunoaffinity column (CitriTest(®)) proved to be clearly superior to SPE with RP(18) material for subsequent analysis by LC-MS/MS. For CIT and its metabolite dihydrocitrinone (HO-CIT), the LOD and LOQ determined by external calibration curves in matrix were 0.02 and 0.05 ng/mL for CIT, and those for HO-CIT were 0.05 and 0.1 ng/mL urine. The newly developed method was applied in a small pilot study: CIT was present in all plasma samples from 8 German adults, at concentrations ranging from 0.11 to 0.26 ng/mL. The molar (nM) concentrations of CIT are similar to those measured for OTA in these samples as a result of dietary mycotoxin intake. CIT was detected in 8/10 urines (from 4 adults and 6 infants) in a range of 0.16-0.79 ng/mL, and HO-CIT was present in 5/10 samples at similar concentrations. Thus, CIT is excreted in urine as parent compound and also as metabolite. These first results in humans point to the need for further studies on CIT exposure.

Topics: Adolescent; Adult; Biomarkers; Biotransformation; Calibration; Chemical Precipitation; Chromatography, High Pressure Liquid; Citrinin; Female; Food Contamination; Food Microbiology; Germany; Humans; Infant; Limit of Detection; Male; Middle Aged; Pilot Projects; Reference Standards; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Young Adult

Leishmaniasis (1) is an endemic disease mainly caused by the protozoan Leishmania donovani (Ld). Polyamines have been identified as essential organic compounds for the growth and survival of Ld. These are synthesized in Ld by polyamine synthesis pathway comprising of many enzymes such as ornithine decarboxylase (ODC), spermidine synthase (SS), and S-adenosylmethionine decarboxylase. Inhibition of these enzymes in Ld offers a viable prospect to check its growth and development. In the present work, we used computational approaches to search natural inhibitors against ODC and SS enzymes. We predicted three-dimensional structures of ODC and SS using comparative modeling and molecular dynamics (MD) simulations. Thousands of natural compounds were virtually screened against target proteins using high throughput approach. MD simulations were then performed to examine molecular interactions between the screened compounds and functional residues of the active sites of the enzymes. Herein, we report two natural compounds of dual inhibitory nature active against the two crucial enzymes of polyamine pathway of Ld. These dual inhibitors have the potential to evolve as lead molecules in the development of antileishmanial drugs. (1)These authors contributed equally.

Topics: Antiprotozoal Agents; Catalytic Domain; Citrinin; Drug Discovery; Enzyme Inhibitors; Heterocyclic Compounds, 3-Ring; Hydrogen Bonding; Hydrophobic and Hydrophilic Interactions; Kinetics; Leishmania donovani; Libraries, Digital; Molecular Dynamics Simulation; Ornithine Decarboxylase; Ornithine Decarboxylase Inhibitors; Polyamines; Protein Binding; Protein Conformation; Protozoan Proteins; Small Molecule Libraries; Spermidine Synthase; Thermodynamics; Tryptophan

To study the secondary metabolites of mangrove endophytic fungus SK5.. The compounds were isolated by chromatographic technique. Their structures were identified by comprehensive physico-chemical properties and spectroscopic methods.. Five compounds were isolated and identified as 2,8-dihydroxy-3,4-dihydronaphthalen-1(2H)-one(1), Sclerotinin A (2), dihydrocitrinone(3), 4-hydroxybenzaldehyde (4) and 3-hydroxy-2-methylbenzoic acid (5).. Compound 1 is isolated from nature for the first time.

Topics: Benzaldehydes; China; Chromatography, High Pressure Liquid; Citrinin; Fungi; Molecular Structure; Naphthols; Rhizophoraceae; Vanillic Acid