ABSTRACT
OBJECTIVES: We have assessed the effect of elevated concentrations of hydroxyphenylpyruvic acid (HPPA), hydroxyphenyllactic acid (HPLA) and tyrosine, on a range of chemistry tests in serum and urine to explore the potential for chemical interference on routine laboratory analyses in patients with alkaptonuria (AKU) treated with nitisinone and similarly implications for patients with hereditary tyrosinemia type 1 (HT-1). MATERIALS AND METHODS: HPPA, HPLA and tyrosine were added separately to pooled serum from subjects without AKU in a range of assays with Roche Modular chemistries. Effects on urine were assessed by changes in urine strip chemistries after mixing a positive control urine with various amounts of the test compounds and reading on a Siemens urine strip meter. RESULTS: No significant effect (pâ¯>â¯0.1) was observed up to 225⯵mol/L of HPPA and HPLA, and up to 5000⯵mol/L tyrosine, on any of the serum-based assays including those with peroxidase-coupled reaction systems of enzymatic creatinine, urate, total cholesterol, HDL cholesterol and triglyceride. Both the monohydroxy HPPA, and the dihydroxy homogentisic acid (HGA), at increased urine concentrations typical of nitisinone-treated AKU and non-treated AKU respectively, did however show marked negative interference in strip assays for glucose and leucocytes; i.e. those with peroxide-linked endpoints. The effect of increased HPLA was less marked. CONCLUSIONS: In patients with AKU or on nitisinone treatment and HT-1 patients on nitisinone, urine strip chemistry testing should be used sparingly, if at all, to avoid false negative reporting. It is recommended that urine assays should be organised with a suitable specialist laboratory.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Alkaptonuria/drug therapy , Alkaptonuria/metabolism , Cyclohexanones/therapeutic use , Enzyme Inhibitors/therapeutic use , Nitrobenzoates/therapeutic use , Phenylpropionates/analysis , Phenylpyruvic Acids/analysis , Tyrosine/metabolism , Alkaptonuria/blood , Alkaptonuria/urine , Humans , Phenylpropionates/blood , Phenylpropionates/urine , Phenylpyruvic Acids/blood , Phenylpyruvic Acids/urineABSTRACT
Loxoprofen is a prodrug that exerts strong analgesic and anti-inflammatory effects through its active trans-alcohol metabolite, which is produced in the liver by carbonyl reductase. Previous metabolic studies have evaluated loxoprofen, but its sulfate and taurine conjugates have not yet been studied. We characterized the metabolomic profile of loxoprofen in rat plasma, urine, and feces using high-resolution mass spectrometry. We identified 17 metabolites of loxoprofen in the three different biological matrices, 13 of which were detected in plasma and feces and 16 in urine. Amongst these metabolites, two novel taurine conjugates (M12 and M13) and two novel acyl glucuronides (M14, M15) were identified for the first time in rats. In addition, we detected three novel sulfate conjugates (M9, M10, and M11) of loxoprofen. Further study of these metabolites of loxoprofen is essential in order to assess their potency and toxicity.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacokinetics , Phenylpropionates/pharmacokinetics , Prodrugs/pharmacokinetics , Administration, Oral , Animals , Anti-Inflammatory Agents, Non-Steroidal/blood , Anti-Inflammatory Agents, Non-Steroidal/urine , Feces/chemistry , Male , Metabolomics , Phenylpropionates/blood , Phenylpropionates/urine , Rats, Sprague-Dawley , Sulfates/metabolismABSTRACT
The focus of the present study is on in vitro and in vivo metabolite identification of ambrisentan (AMBR) a selective endothelin type - A (ETA) receptor antagonist using quadruple time-of-flight mass spectrometry (QTOF/MS). in vitro metabolism study was conducted by incubating AMBR in rat liver microsomes (RLM), rat and human liver S9 fractions. In vivo study was carried out through the collection of urine, faeces and plasma samples at various time points after oral administration of AMBR in suspension form at a dose of 25â¯mg/kg to six male Sprague - Dawley (SD) rats. The samples were prepared using an optimized sample preparation techniques involving protein precipitation (PP), freeze liquid extraction (FLE) and solid phase extraction (SPE). The extracted samples were further concentrated and analyzed by developing a sensitive and specific liquid chromatography-mass spectrometry (LC-MS) method. A total of seventeen metabolites were identified in in vivo samples which includes hydroxyl, demethylated, demethoxylated, hydrolytic, decarboxylated, epoxide and glucuronide metabolites. Most of the metabolites were observed in faeces and urine matrices and few were observed in the plasma matrix. Only ten metabolites were identified in in vitro study which was commonly observed in in vivo study. The detailed structural elucidation of all the metabolites was done using UHPLC/QTOF/MS/MS in combination with accurate mass measurements. The toxicity profile of AMBR and its metabolites were predicted using TOPKAT software. In addition, a mass spectrometric method was developed for the detection and characterization of GSH-trapped reactive epoxide metabolitein human liver S9 fraction supplemented with glutathione (GSH) as trapping agent.
Subject(s)
Epoxy Compounds/chemistry , Epoxy Compounds/metabolism , Glutathione/blood , Phenylpropionates/chemistry , Phenylpropionates/metabolism , Pyridazines/chemistry , Pyridazines/metabolism , Animals , Chromatography, High Pressure Liquid/methods , Feces/chemistry , Glutathione/metabolism , Humans , Male , Microsomes, Liver/metabolism , Phenylpropionates/blood , Phenylpropionates/urine , Plasma/chemistry , Pyridazines/blood , Pyridazines/urine , Rats , Rats, Sprague-Dawley , Software , Solid Phase Extraction/methods , Spectrometry, Mass, Electrospray Ionization/methods , Tandem Mass Spectrometry/methods , Urine/chemistryABSTRACT
Loxoprofen is a non-steroidal anti-inflammatory drug of the 2-arylpropionic acid type, which has used to treat musculoskeletal disorders in the horse racing industry. However, it has also used illicitly to mask clinical signs of inflammation and pain in racehorses. Thus, its accurate analysis has become an important issue in horse doping laboratories. In this study, an analytical method of loxoprofen was developed as tert-butyldimethylsilyl (TBDMS) derivative by gas chromatography-mass spectrometry (GC-MS). Characteristic fragment ions of [M-15], [M-57], and [M-139] permitted the accurate and selective detection of loxoprofen. Under optimal conditions, this method showed good linearity (r ≥ 0.999) in the range of 10-500 ng/mL, repeatability (% relative standard deviation = 5.6-8.5), and accuracy (% relative error = - 0.3-0.9) with a detection limit of 1.0 ng. When applied to the analysis of loxoprofen in tablet and patch products, loxoprofen was positively identified as TBDMS derivative by GC-MS. The present method provided rapid and accurate determination of loxoprofen in patch and tablet products. Levels of loxoprofen were highest in equine urine at 0.5 and 1 h after oral administration with single dose (3 mg/kg) to three horses, and then rapidly reduced to below the lower limit of quantification at 24 h. Therefore, the present method will be useful for the pharmacokinetic study and doping tests for loxoprofen and other similar acidic drugs in horses.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/analysis , Gas Chromatography-Mass Spectrometry/methods , Organosilicon Compounds/analysis , Phenylpropionates/analysis , Tablets/analysis , Transdermal Patch , Administration, Oral , Animals , Anti-Inflammatory Agents, Non-Steroidal/urine , Horses , Phenylpropionates/urineABSTRACT
BACKGROUND: In severe falciparum malaria metabolic acidosis and acute kidney injury (AKI) are independent predictors of a fatal outcome in all age groups. The relationship between plasma acids, urine acids and renal function was investigated in adult patients with acute falciparum malaria. METHODS: Plasma and urinary acids which previously showed increased concentrations in proportion to disease severity in patients with severe falciparum malaria were quantified. Patients with uncomplicated malaria, sepsis and healthy volunteers served as comparator groups. Multiple regression and multivariate analysis were used to assess the relationship between organic acid concentrations and clinical syndromes, in particular AKI. RESULTS: Patients with severe malaria (n = 90), uncomplicated malaria (n = 94), non-malaria sepsis (n = 19), and healthy volunteers (n = 61) were included. Univariate analysis showed that both plasma and creatinine-adjusted urine concentrations of p-hydroxyphenyllactic acid (pHPLA) were higher in severe malaria patients with AKI (p < 0.001). Multiple regression analysis, including plasma or creatinine-adjusted urinary acids, and PfHRP2 as parasite biomass marker as independent variables, showed that pHPLA was independently associated with plasma creatinine (ß = 0.827) and urine creatinine (ß = 0.226). Principal component analysis, including four plasma acids and seven urinary acids separated a group of patients with AKI, which was mainly driven by pHPLA concentrations. CONCLUSIONS: Both plasma and urine concentrations of pHPLA closely correlate with AKI in patients with severe falciparum malaria. Further studies will need to assess the potential nephrotoxic properties of pHPLA.
Subject(s)
Acidosis/metabolism , Acute Kidney Injury/metabolism , Malaria, Falciparum/complications , Phenylpropionates/blood , Phenylpropionates/urine , Sepsis/complications , Acidosis/parasitology , Acidosis/physiopathology , Acids/blood , Acids/urine , Acute Kidney Injury/parasitology , Acute Kidney Injury/physiopathology , Adult , Bangladesh , Female , Humans , Male , Middle Aged , Young AdultABSTRACT
SCOPE: Most studies on the role of whole grain for health rely on self-reported intake data, which are prone to measurement errors. There is a need for dietary biomarkers that can provide an objective measure of intake. Alkylresorcinols (AR) and their main metabolites 3,5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-propanoic acid (DHPPA) have been proposed as biomarkers for whole grain (WG) wheat and rye intake. METHODS AND RESULTS: The medium-term reproducibility and relative validity of four putative urinary AR metabolites (3,5-dihydroxycinnamic acid (DHCA), 5-(3,5-dihydroxyphenyl) pentanoic acid (DHPPTA), 2-(3,5-dihydroxybenzamido)acetic acid (DHBA-glycine) and 3,5-dihydroxycinnamic acid amide (DHCA-amide)) as biomarkers for WG intake were investigated. Three-day weighed food records and 24-h urine samples from two occasions 2-3 months apart were obtained from 69 Swedish adults. WG intake was calculated and urinary AR metabolites were analyzed. The medium-term reproducibility determined for DHCA, DHPPTA, and DHBA-glycine varied from moderate-to-excellent (intra-class correlation coefficient = 0.63-0.85). Moreover, DHCA and DHPPTA excretion correlated well with self-reported total WG intake (r = 0.55, p < 0.001 and r = 0.42, p < 0.001, respectively). CONCLUSION: DHCA or DHPPTA excretion in 24-h urine might be a suitable medium- to long-term biomarker of WG wheat and rye intake. These findings need to be confirmed in populations with low and infrequent WG intake.
Subject(s)
Cinnamates/urine , Phenylpropionates/urine , Resorcinols/pharmacokinetics , Whole Grains , Adult , Biomarkers/urine , Female , Humans , Hydroxybenzoates/urine , Male , Middle Aged , Reproducibility of Results , Resorcinols/metabolism , Resorcinols/urine , Secale , Sweden , TriticumABSTRACT
Autism spectrum disorders (ASDs) are a group of mental illnesses highly correlated with gut microbiota. Recent studies have shown that some abnormal aromatic metabolites in autism patients are presumably derived from overgrown Clostridium species in gut, which may be used for diagnostic purposes. In this paper, a GC/MS based metabolomic approach was utilized to seek similar biomarkers by analyzing the urinary information in 62 ASDs patients compared with 62 non-ASDs controls in China, aged 1.5-7. Three compounds identified as 3-(3-hydroxyphenyl)-3-hydroxypropionic acid (HPHPA), 3-hydroxyphenylacetic acid (3HPA), and 3-hydroxyhippuric acid (3HHA) were found in higher concentrations in autistic children than in the controls (p < 0.001). After oral vancomycin treatment, urinary excretion of HPHPA (p < 0.001), 3HPA (p < 0.005), and 3HHA (p < 0.001) decreased markedly, which indicated that these compounds may also be from gut Clostridium species. The sensitivity and specificity of HPHPA, 3HPA, and 3HHA were evaluated by receiver-operating characteristic (ROC) analysis. The specificity of each compound for ASDs was very high (>96%). After two-regression analysis, the optimal area under the curve (AUC, 0.962), sensitivity (90.3%), and specificity (98.4%) were obtained by ROC curve of Prediction probability based on the three metabolites. These findings demonstrate that the measurements of the three compounds are strong predictors of ASDs and support the potential clinical utility for identifying a subgroup of ASDs subjects.
Subject(s)
Autism Spectrum Disorder/epidemiology , Autism Spectrum Disorder/urine , Hippurates/urine , Phenylacetates/urine , Phenylpropionates/urine , Autism Spectrum Disorder/diagnosis , Biomarkers/urine , Child , Child, Preschool , China/epidemiology , Female , Humans , Infant , Male , Prevalence , Reproducibility of Results , Risk Factors , Sensitivity and SpecificityABSTRACT
PURPOSE: Alkylresorcinols (AR) are phenolic lipids present in the bran of wheat and rye. Plasma AR and their urinary metabolites may be suitable biomarkers of whole-grain (WG) wheat and rye consumption. The objective of this study was to examine plasma AR and urinary AR metabolites in response to WG wheat consumption. METHODS: In a randomized crossover study, 19 subjects (10 males, 9 females; BMI 22.0 kg/m(2); age 26 years) incorporated either 3 servings (48 g) or 6 servings (96 g) of WG wheat daily into their regular diet for 1 week. Subjects completed a 2-week washout period, abstaining from all WG consumption, before each intervention. Fasting blood and 24-h urine were collected before and after each intervention. Plasma AR homologues (C19:0, C21:0, C23:0) were quantified by GC-MS after diethyl ether and solid phase extraction and derivatization. Urinary AR metabolites [3,5-dihydroxybenzoic acid and 3-(3,5-dihydroxyphenyl)-propanoic acid] were determined using HPLC with electrochemical detection after enzymatic deconjugation and ethyl acetate extraction. RESULTS: Urinary total AR metabolites were significantly higher after 6 compared with 3 servings of WG wheat (56 vs. 32 µmol/day, P < 0.001). This dose-response relationship was independent of age, sex, energy intake, and baseline urinary AR metabolite concentration. Plasma total AR tended to be higher after 6 compared with 3 servings of WG wheat (103.0 vs. 86.9 nmol/L), but this difference was not significant (P = 0.42). CONCLUSION: The results suggest that urinary AR metabolites from 24-h urine collections may be useful as biomarkers of compliance in intervention studies of WG wheat.
Subject(s)
Biomarkers/blood , Biomarkers/urine , Diet , Patient Compliance , Resorcinols/chemistry , Whole Grains , Adolescent , Adult , Body Mass Index , Cross-Over Studies , Dietary Fiber/administration & dosage , Female , Humans , Hydroxybenzoates/urine , Male , Phenylpropionates/urine , Resorcinols/urine , Secale , Triticum , Young AdultABSTRACT
Alkylresorcinols (ARs) are phytochemicals mainly associated with rye/wheat bran. Plasma ARs and their plasma and urine metabolites are considered as biomarkers for whole-grain rye/wheat intake. However ARs metabolite day and night variations have not been studied in prostate cancer patients yet. We investigated ARs metabolites 3, 5-dihydroxy-benzoic acid (DHBA), and 3-(3, 5-dihydroxyphenyl)-1-propanoic acid (DHPPA) in urine and plasma in prostate cancer patients and in control group. DHPPA in 12-h overnight urine correlated with the intake of rye bread and bread fiber across short time periods (3 days). Plasma DHPPA concentration was significantly greater in the prostate cancer group than in the control group. DHPPA and DHBA excretion was significantly higher in the overnight urine than in day urine in the prostate cancer group but not in the control group. DHPPA concentration in plasma in the prostate cancer group did not depend on the intake of rye bread in the previous day, suggesting an impaired metabolism of ARs metabolites in the prostate cancer group. The results of this study suggest DHPPA in 12-h overnight urine as a biomarker to estimate the intake of rye bread and bread fiber.
Subject(s)
Hydroxybenzoates/blood , Hydroxybenzoates/urine , Phenylpropionates/blood , Phenylpropionates/urine , Prostatic Neoplasms/blood , Prostatic Neoplasms/urine , Resorcinols/blood , Resorcinols/urine , Secale/metabolism , Triticum/metabolism , Aged , Biomarkers/blood , Biomarkers/urine , Bread , Case-Control Studies , Circadian Rhythm , Dietary Fiber/metabolism , Humans , Male , Middle AgedABSTRACT
The aim of this study was to investigate the relation between the etiology of late-onset childhood autism and anaerobic bacteria. Thirty children diagnosed with autistic disorder and control group have been included in the study. 3-(3-hydroxy phenyl)-3-hydroxypropionic acid (HPHPA) excretion rates which is a metabolic product of the genus Clostridium, were measured via mass spectrometry-gas chromatography (MS-GC) method from urine samples. When the assayed average HPHPA values compared with each group, a statistically significant difference was found (p < 0.05). Data obtained from this study support the existence of a significant correlation between autism etiology and anaerobic bacteria.
Subject(s)
Autistic Disorder/diagnosis , Clostridium/metabolism , Phenylpropionates , Adolescent , Age of Onset , Anaerobiosis , Autistic Disorder/epidemiology , Autistic Disorder/microbiology , Autistic Disorder/urine , Case-Control Studies , Child , Child, Preschool , Clostridium/pathogenicity , Female , Gas Chromatography-Mass Spectrometry , Humans , Male , Phenylpropionates/urine , Turkey/epidemiologyABSTRACT
Biomarkers of dietary intake are prominent tools in nutritional research. The alkylresorcinol metabolites 3,5-dihydroxybenzoic acid (3,5-DHBA) and 3-(3,5-dihydroxyphenyl)propanoic acid (3,5-DHPPA) have been proposed as exposure biomarkers of whole-grain (WG) wheat and rye intake. However, the profile of alkylresorcinol metabolites is not fully understood. The aim of this study was to investigate the metabolism of alkylresorcinols in mice and in humans, while further determining urinary pharmacokinetics of the novel alkylresorcinol metabolites to explore their potential as biomarkers of WG wheat intake. Utilization of the liquid chromatography-mass spectrometry approach resulted in 10 alkylresorcinol metabolites identified in mice and in humans, including 3 phenolic acids and 7 of their phase II conjugates. Among them, 2 novel metabolites were discovered: 5-(3,5-dihydroxyphenyl)pentanoic acid (3,5-DHPPTA) and 2-(3,5-dihydroxybenzamido)acetic acid (3,5-DHBA glycine). The structures of these 2 metabolites were confirmed by comparing with authentic standards synthesized in-house. In the pharmacokinetic study, a group of 12 volunteers consumed a polyphenolic-restricted diet for 4 d before ingesting WG wheat bread containing 61 mg of alkylresorcinols. Urine samples were collected for 32 h, and alkylresorcinol metabolites were quantified with HPLC-coulometric electrode array detection. The mean urinary excretion rates and mean apparent half-life of 3,5-DHPPTA, 3,5-DHBA glycine, 3,5-DHBA, and 3,5-DHPPA at each time point were determined. Our results suggest that 3,5-DHPPTA and 3,5-DHBA glycine may be used in combination with 3,5-DHBA and 3,5-DHPPA as potential biomarkers to increase the accuracy of recording WG wheat and rye intake in epidemiologic studies. Further validation of 3,5-DHPPTA and 3,5-DHBA glycine as potential biomarkers is warranted.
Subject(s)
Biomarkers/urine , Diet , Plant Preparations/pharmacokinetics , Resorcinols/urine , Secale , Triticum , Acetates/metabolism , Acetates/urine , Adult , Animals , Chromatography, High Pressure Liquid , Edible Grain , Female , Humans , Hydroxybenzoates/metabolism , Hydroxybenzoates/urine , Male , Mice , Mice, Inbred C57BL , Molecular Structure , Pentanoic Acids/metabolism , Pentanoic Acids/urine , Phenylpropionates/metabolism , Phenylpropionates/urine , Plant Preparations/metabolism , Polyphenols/administration & dosage , Resorcinols/metabolism , SeedsABSTRACT
SCOPE: Non-targeted urine metabolite profiling has not been previously exploited in the field of whole grain (WG) products. WG products, particularly rye, are important elements in a healthy Nordic diet. The aim of this study was to identify novel urinary biomarkers of WG rye bread (RB) intake in a randomised crossover study with RB versus refined wheat bread (WB). METHODS AND RESULTS: UPLC-QTOF/MS metabolite profiling was applied to urine from a 2 × 4 wk crossover intervention with RB versus WB in 20 subjects. Sixteen metabolites were revealed as major contributing biomarkers. The most discriminative metabolite after the cereal intervention was identified as 3-(3,5-dihydroxyphenyl)-1-propanoic acid sulphate, which was excreted to a higher extent after the RB versus WB intervention. Other alkylresorcinol metabolites were identified, as well as enterolactone glucuronide, azelaic acid, 2-aminophenol sulphate and its benzoxazinoid precursor 2,4-dihydroxy-1,4-benzoxazin-3-one. Our study also suggests that nitrogen-containing metabolites are other major markers. However, other methodologies will be needed to elucidate their final structure. CONCLUSION: The present non-targeted metabolite profiling proved to be a useful approach to identify major urine metabolites discriminating RB intake from that of white wheat bread. Once validated these markers could help evaluate compliance to healthy Nordic diets.
Subject(s)
Biomarkers/urine , Bread , Diet , Secale , Triticum , 4-Butyrolactone/analogs & derivatives , 4-Butyrolactone/urine , Adult , Aminophenols/urine , Benzoxazines/urine , Cross-Over Studies , Female , Humans , Lignans/urine , Male , Mass Spectrometry/methods , Metabolome , Middle Aged , Phenylpropionates/urineABSTRACT
BACKGROUND: The enzyme 4-hydroxyphenylpyruvate dioxygenase (HPPD) is key in tyrosine catabolism. Inhibition of HPPD results in tyrosinemia and increased urinary excretion of 3 phenylketones: 4-hydroxyphenylpyruvate (HPPA), 4-hydroxyphenyllactate (HPLA), and 4-hydroxyphenylacetate (HPAA). A previous study involving administration of a novel HPPD inhibitor to dogs resulted in detection of ketonuria in treated animals using urine dipsticks read by reflectance photometry. Dipstick-positive results were suspected to be false because high concentrations of urinary phenylketones have been reported to react with ketone test fields of urine dipsticks, but visual confirmation was not performed. OBJECTIVE: The purpose of this study was to determine which of the 4- hydroxyphenolic acids produced by HPPD inhibition react with ketone test fields of 3 commercially available urine dipsticks. METHODS: Canine urine samples were prepared with HPPA, HPLA, HPAA, and lithium acetoacetate (positive control) at 6 concentrations. Unmodified urine samples were used as negative controls. All samples were tested for ketones using Combur 10 Test M dipsticks read by a Miditron dipstick analyzer. Urinalysis was also performed by visually inspecting ketone test fields on the Combur 10 Test M, Multistix 10 SG, and Aution 10 EA dipsticks. RESULTS: Urine samples containing HPPA were positive for ketones with Combur 10 Test M dipsticks read by the Miditron analyzer and produced a redbrown color change in ketone test fields of all 3 dipsticks. Urine samples containing HPLA and HPAA were negative by all methods. CONCLUSION: The phenylketone HPPA reacts with ketone test fields of 3 commercially available urine dipsticks, producing a redbrown color change that may be misinterpreted as positive for ketones by reflectance photometry.
Subject(s)
Dogs/urine , Ketones/urine , Phenylpyruvic Acids/metabolism , Tyrosine/metabolism , Animals , Dog Diseases/diagnosis , Dog Diseases/metabolism , Dog Diseases/urine , Dogs/metabolism , False Positive Reactions , Female , Ketosis/diagnosis , Ketosis/metabolism , Ketosis/urine , Male , Phenylacetates/metabolism , Phenylacetates/urine , Phenylpropionates/metabolism , Phenylpropionates/urine , Phenylpyruvic Acids/urine , Reagent Kits, Diagnostic/standards , Reagent Kits, Diagnostic/veterinary , Reagent Strips , Tyrosine/urineABSTRACT
A compound identified as 3-(3-hydroxyphenyl)-3-hydroxypropionic acid (HPHPA) was found in higher concentrations in urine samples of children with autism compared to age and sex appropriate controls and in an adult with recurrent diarrhea due to Clostridium difficile infections. The highest value measured in urine samples was 7500 mmol/mol creatinine, a value 300 times the median normal adult value, in a patient with acute schizophrenia during an acute psychotic episode. The psychosis remitted after treatment with oral vancomycin with a concomitant marked decrease in HPHPA. The source of this compound appears to be multiple species of anaerobic bacteria of the Clostridium genus. The significance of this compound is that it is a probable metabolite of m-tyrosine (3-hydroxyphenylalanine), a tyrosine analog which depletes brain catecholamines and causes symptoms of autism (stereotypical behavior, hyperactivity, and hyper-reactivity) in experimental animals.
Subject(s)
Autistic Disorder/urine , Gastrointestinal Tract/microbiology , Phenylpropionates/urine , Schizophrenia/urine , Acute Disease , Adolescent , Adult , Child , Child, Preschool , Clostridioides difficile , Clostridium/metabolism , Clostridium Infections/complications , Clostridium Infections/drug therapy , Enterocolitis, Pseudomembranous/microbiology , Enterocolitis, Pseudomembranous/urine , Female , Humans , Male , Middle Aged , Phenylalanine/metabolism , Schizophrenia/complications , Sex Characteristics , Vancomycin/therapeutic use , Young AdultABSTRACT
BACKGROUND: Hereditary tyrosinemia type 1 (HT1; MIM 276700) is caused by mutations in the fumarylaceto-acetate hydrolase (FAH) gene, and is the most severe disorder associated with the tyrosine catabolic pathway. HT1 is a very rare disorder and no genetically confirmed case of HT1 in Korea has yet been reported. In this study, we present a Korean neonate with clinical and biochemical features of HT1. METHODS: A female neonate was admitted to our hospital for further work-up of an abnormal newborn screening test. We analyzed amino acids and organic acids in the patient's blood and urine. To confirm the presence of the genetic abnormality, all the coding exons of the FAH gene and the flanking introns were amplified by polymerase chain reaction (PCR). RESULTS: The patient's newborn screening test revealed increased concentrations of methionine and tyrosine. Subsequent urine organic acid analysis showed increased urinary excretion of 4-hydroxyphenyllactate, 4-hydroxyphenylpyruvate, succinate, and succinylacetone. Gap-PCR and sequence analysis of the FAH gene revealed a homozygous large deletion mutation encompassing exons 12-14. The patient's parents were not consanguineous but were heterozygous carriers of the same mutation. CONCLUSIONS: The patient had a novel, large deletion mutation of FAH and is the first report of genetically confirmed HT1 in Korea.
Subject(s)
Hydrolases/genetics , Tyrosinemias/genetics , Exons/genetics , Female , Heptanoates/urine , Humans , Hydrolases/blood , Hydrolases/urine , Infant, Newborn , Introns/genetics , Phenylpropionates/urine , Phenylpyruvic Acids/urine , Sequence Deletion/genetics , Succinic Acid/urine , Tyrosinemias/metabolismABSTRACT
In vivo and in vitro metabolism of scopolamine is investigated using a highly specific and sensitive liquid chromatography-mass spectrometry (LC-MSn) method. Feces, urine, and plasma samples are collected individually after ingestion of 55 mg/kg scopolamine by healthy rats. Rat feces and urine samples are cleaned up by a liquid-liquid extraction and a solid-phase extraction procedure (C18 cartridges), respectively. Methanol is added to rat plasma samples to precipitate plasma proteins. Scopolamine is incubated with homogenized liver and intestinal flora of rats in vitro, respectively. The metabolites in the incubating solution are extracted with ethyl acetate. Then these pretreated samples are injected into a reversed-phase C18 column with mobile phase of methanol-ammonium acetate (2 mM, adjusted to pH 3.5 with formic acid) (70:30, v/v) and detected by an on-line MSn system. Identification and structural elucidation of the metabolites are performed by comparing their changes in molecular masses (DeltaM), retention-times and full scan MSn spectra with those of the parent drug. The results reveal that at least 8 metabolites (norscopine, scopine, tropic acid, aponorscopolamine, aposcopolamine, norscopolamine, hydroxyscopolamine, and hydroxyscopolamine N-oxide) and the parent drug exist in feces after administering 55 mg/kg scopolamine to healthy rats. Three new metabolites (tetrahydroxyscopolamine, trihydroxy-methoxyscopolamine, and dihydroxy-dimethoxyscopolamine) are identified in rat urine. Seven metabolites (norscopine, scopine, tropic acid, aponorscopolamine, aposcopolamine, norscopolamine, and hydroxyscopolamine) and the parent drug are detected in rat plasma. Only 1 hydrolyzed metabolite (scopine) is found in the rat intestinal flora incubation mixture, and 2 metabolites (aposcopolamine and norscopolamine) are identified in the homogenized liver incubation mixture.
Subject(s)
Chromatography, High Pressure Liquid/methods , Scopolamine/analysis , Spectrometry, Mass, Electrospray Ionization/methods , Animals , Molecular Structure , Phenylpropionates/analysis , Phenylpropionates/blood , Phenylpropionates/urine , Rats , Rats, Wistar , Reproducibility of Results , Scopolamine/blood , Scopolamine/urine , Scopolamine Derivatives/analysis , Scopolamine Derivatives/blood , Scopolamine Derivatives/urineABSTRACT
Three phase liquid phase microextraction (three phase LPME) technique coupled with HPLC-UV has been applied as a sensitive and efficient sample preparation method to determine phenylacetic acid (PAA) as a biomarker of depressive disorders and phenylpropionic acid (PPA) in biological fluids. The compounds were extracted from 3.0 ml aqueous solution with the adjustment of pH at a fixed value in the range of 2.0-3.5 (donor solution) into an organic phase (1-hexanol) layered on the surface of the donor solution and finally back-extracted into 4.0 microl of the acceptor microdrop (pH 11.1) located at the end of the microsyringe needle. After a prescribed back-extraction time, the acceptor microdrop was withdrawn into the microsyringe and then directly injected into the HPLC system. In order to achieve maximum extraction efficiency, different parameters affecting the extraction conditions were optimized. At the optimum conditions (donor solution: 2.3M Na(2)SO(4), pH 2.0-3.5; organic membrane: 95 microl of 1-hexanol; acceptor solution: 4.0 microl of 0.1M NH(3)/NH(4)(+) with pH 11.1; donor solution temperature: 45-50 degrees C; extraction time: 20 min and back-extraction time: 12 min), up to 110-fold enrichment factor was obtained. The calibration curve for these analytes was linear in the range of 1-5000 microg/l with r(2)>0.998. The intraday and interday RSD% were below 6.5% and the limits of detection (LODs) for both analytes were 0.2 microg/l (based on S/N=3). The proposed technique is a low cost, simple and sensitive method with highly clean-up effect. Finally, this technique was successfully utilized for the detection of target analytes in human urine, serum and plasma.
Subject(s)
Chromatography, High Pressure Liquid/methods , Phenylacetates/isolation & purification , Phenylpropionates/isolation & purification , Humans , Hydrogen-Ion Concentration , Phenylacetates/blood , Phenylacetates/urine , Phenylpropionates/blood , Phenylpropionates/urine , TemperatureABSTRACT
Naveglitazar [LY519818; benzenepropanoic acid, alpha-methoxy-4-[3-(4-phenoxyphenoxy)propoxy], (alpha-S)-] is a nonthiozolidinedione peroxisome proliferator-activated receptor alpha-gamma dual, gamma-dominant agonist that has shown glucose-lowering potential in animal models and in the clinic. Studies have been conducted to characterize the disposition, metabolism, and excretion of naveglitazar in mice, rats, and monkeys after oral and/or i.v. bolus administration. After oral administration of [(14)C]naveglitazar, naveglitazar was well absorbed and moderately metabolized in all species evaluated, with total recoveries of radioactivity ranging from 90 to 96%. Naveglitazar was the most abundant peak observed in circulation at C(max), representing 68 to 81% of the total radioactivity in plasma. The most prominent metabolite observed in circulation was the R-enantiomer of naveglitazar, LY591026, which is formed via enzymatic chiral inversion. para-Hydroxy naveglitazar and the sulfate conjugate of para-hydroxy naveglitazar were also observed in circulation in most species, especially in the monkey. The metabolic pathways observed include enzymatic chiral inversion, aromatic hydroxylation, oxidative dehydrogenation, and/or various phase II conjugation pathways. Naveglitazar was highly bound to plasma proteins among the species examined (>99%), and binding was independent of concentration. Biliary excretion was recognized as the most prominent excretion pathway in bile duct-cannulated rats (79 of the 96% recovered), producing an acyl glucuronide conjugate of naveglitazar and a sulfate and glucuronide diconjugate of para-hydroxy naveglitazar, which were shown to be reversible. The primary excretory pathway observed in mice and monkeys was via the feces. In summary, naveglitazar was well absorbed, moderately metabolized, and excreted via the feces in mice, rats, and monkeys.
Subject(s)
Hypoglycemic Agents/pharmacokinetics , PPAR alpha/agonists , PPAR gamma/agonists , Phenylpropionates/pharmacokinetics , Animals , Bile/chemistry , Blood Proteins/metabolism , Feces/chemistry , Humans , Hypoglycemic Agents/blood , Hypoglycemic Agents/urine , Liver/metabolism , Macaca fascicularis , Mice , Mice, Inbred ICR , Phenylpropionates/blood , Phenylpropionates/urine , Protein Binding , Rats , Rats, Inbred F344Subject(s)
Keratoderma, Palmoplantar/etiology , Tyrosinemias/complications , Adult , Biomarkers/blood , Biomarkers/urine , Humans , Keratoderma, Palmoplantar/diagnosis , Keratoderma, Palmoplantar/metabolism , Male , Phenylacetates/blood , Phenylacetates/urine , Phenylpropionates/blood , Phenylpropionates/urine , Phenylpyruvic Acids/blood , Phenylpyruvic Acids/urine , Tyrosine/blood , Tyrosine/urine , Tyrosinemias/diagnosis , Tyrosinemias/metabolismABSTRACT
The effect of pretreatment (i.e., oral administration of loxoprofen for 3 consecutive days followed by a 7-day washout) on the pharmacokinetics and metabolism of the drug was studied in humans. In a control study, a Loxonin tablet (60 mg as loxoprofen anhydrous) was administered orally to 6 healthy male Korean subjects. In a pretreatment study, a Loxonin tablet was administered orally to the subjects once daily for 3 consecutive days. On the 10(th) day, a Loxonin tablet was administered orally to the subjects, and the concentrations of loxoprofen and the trans- and cis-alcohol metabolites in the plasma and urine were measured as a function of time. Using this pretreatment, the area under the curve (AUC) of the trans-alcohol metabolite of loxoprofen in the plasma, but not those of loxoprofen and the cis-alcohol metabolite, was increased (1.5-fold, p < 0.05), leading to increased contribution of the trans-alcohol metabolite to the total urinary recovery of loxoprofen (1.3-fold, p < 0.05). The urinary recovery of total metabolites, which was largely (> 90%) comprised of conjugate metabolites, was also increased as a result of the pretreatment (1.5-fold, p < 0.05). These results indicate that stereoselective reduction to trans-alcohol metabolites as well as the phase II metabolism of loxoprofen may be increased by such a pretreatment in human subjects.