ABSTRACT
The preoccupation concerning glyphosate (GLYP) has rapidly grown over recent years, and the availability of genetically modified crops that are resistant to GLYP or glufosinate (GLUF) has increased the use of these herbicides. The debate surrounding the carcinogenicity of GLYP has raised interest and the desire to gain information on the level of exposure of the population. GLYP and aminomethylphosphonic acid (AMPA) are commonly simultaneously analysed. GLUF is sometimes also monitored, but its major metabolite, 3-[hydroxy(methyl)phosphinoyl]propionic acid (3MPPA), is rarely present in the method. Using a pentafluorobenzyl derivative to extract the analytes from human urine, we present a method that contains four important analytes to monitor human exposure to GLYP and GLUF. The use of the flash freeze technique speeds up the extraction process and requires less organic solvent than conventional liquid-liquid extraction. The limits of detection in the low µg/L range enable the use of this method for epidemiological studies. The results obtained for 35 volunteers from the Quebec City area are presented with the results from multiple interlaboratory comparisons (G-EQUAS, HBM4EU and OSEQAS). This methodology is currently being used in the Maternal-Infant Research on Environmental Chemicals (MIREC-ENDO) study and in the Canadian Health Measures Survey (CHMS).
Subject(s)
Aminobutyrates/urine , Chromatography, High Pressure Liquid/methods , Glycine/analogs & derivatives , Herbicides/urine , Tandem Mass Spectrometry/methods , Aminobutyrates/metabolism , Glycine/metabolism , Glycine/urine , Herbicides/metabolism , Humans , Limit of Detection , GlyphosateABSTRACT
Aminomethylphosphonic acid (AMPA) is the main metabolite of glyphosate (GLYP) and phosphonic acids in detergents. GLYP is a synthetic herbicide frequently used worldwide alone or together with its analog glufosinate (GLUF). The general public can be exposed to these potentially harmful chemicals; thus, sensitive methods to monitor them in humans are urgently required to evaluate health risks. We attempted to simultaneously detect GLYP, AMPA, and GLUF in human urine by high-resolution accurate-mass liquid chromatography mass spectrometry (HRAM LC-MS) before and after derivatization with 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) or 1-methylimidazole-sulfonyl chloride (ImS-Cl) with several urine pre-treatment and solid phase extraction (SPE) steps. Fmoc-Cl derivatization achieved the best combination of method sensitivity (limit of detection; LOD) and accuracy for all compounds compared to underivatized urine or ImS-Cl-derivatized urine. Before derivatization, the best steps for GLYP involved 0.4 mM ethylenediaminetetraacetic acid (EDTA) pre-treatment followed by SPE pre-cleanup (LOD 37 pg/mL), for AMPA involved no EDTA pre-treatment and no SPE pre-cleanup (LOD 20 pg/mL) or 0.2-0.4 mM EDTA pre-treatment with no SPE pre-cleanup (LOD 19-21 pg/mL), and for GLUF involved 0.4 mM EDTA pre-treatment and no SPE pre-cleanup (LOD 7 pg/mL). However, for these methods, accuracy was sufficient only for AMPA (101-105%), while being modest for GLYP (61%) and GLUF (63%). Different EDTA and SPE treatments prior to Fmoc-Cl derivatization resulted in high sensitivity for all analytes but satisfactory accuracy only for AMPA. Thus, we conclude that our HRAM LC-MS method is suited for urinary AMPA analysis in cross-sectional studies.
Subject(s)
Aminobutyrates/urine , Chromatography, Liquid/methods , Glycine/analogs & derivatives , Herbicides/urine , Mass Spectrometry/methods , Organophosphonates/urine , Glycine/urine , Humans , Limit of Detection , Reproducibility of Results , Solid Phase Extraction/methods , GlyphosateABSTRACT
AIMS: Sacubitril/valsartan is indicated for the treatment of heart failure and reduced ejection fraction (HFrEF). Furosemide, a loop diuretic commonly used for the treatment of HFrEF, may be coadministered with sacubitril/valsartan in clinical practice. The effect of sacubitril/valsartan on the pharmacokinetics and pharmacodynamics of furosemide was evaluated in this open label, two-period, single-sequence study in healthy subjects. METHODS: All subjects (n = 28) received 40 mg oral single-dose furosemide during period 1, followed by a washout of 2 days. In period 2, sacubitril/valsartan 200 mg (97/103 mg) was administered twice daily for 5 days and a single dose of 40 mg furosemide was coadministered on day 6. Serial plasma and urine samples were collected to determine the pharmacokinetics of furosemide and sacubitril/valsartan and the pharmacodynamics of furosemide. The point estimates and the associated 90% confidence intervals for pharmacokinetic parameters were evaluated. RESULTS: Coadministration of furosemide with sacubitril/valsartan decreased the maximum observed plasma concentration (Cmax ) [estimated geometric mean ratio (90% confidence interval): 0.50 (0.44, 0.56)], area under the plasma concentration-time curve (AUC) from time 0 to infinity [0.72 (0.67, 0.77)] and 24-h urinary excretion of furosemide [0.74 (0.69, 0.79)]. When coadministered with sacubitril/valsartan, 0-4-h, 4-8-h and 0-24-h diuresis in response to furosemide was reduced by ~7%, 21% and 0.2%, respectively, while natriuresis was reduced by ~ 28.5%, 7% and 15%, respectively. Post hoc analysis of the pivotal phase III Prospective comparison of ARNI with ACEI to Determine Impact on Global Mortality and morbidity in Heart Failure trial (PARADIGM-HF) indicated that the median furosemide dose was similar at baseline and at the end of the study in the sacubitril/valsartan group. CONCLUSIONS: Sacubitril/valsartan reduced plasma Cmax and AUC and 24-h urinary excretion of furosemide, while not significantly affecting its pharmacodynamic effects in healthy subjects.
Subject(s)
Aminobutyrates/pharmacology , Aminobutyrates/pharmacokinetics , Drug Interactions , Furosemide/pharmacology , Furosemide/pharmacokinetics , Tetrazoles/pharmacology , Tetrazoles/pharmacokinetics , Adolescent , Adult , Aminobutyrates/blood , Aminobutyrates/urine , Angiotensin Receptor Antagonists/blood , Angiotensin Receptor Antagonists/pharmacokinetics , Angiotensin Receptor Antagonists/pharmacology , Angiotensin Receptor Antagonists/urine , Biphenyl Compounds , Clinical Trials as Topic/statistics & numerical data , Diuresis/drug effects , Diuretics/blood , Diuretics/pharmacokinetics , Diuretics/pharmacology , Diuretics/urine , Drug Combinations , Female , Furosemide/blood , Furosemide/urine , Healthy Volunteers , Humans , Male , Middle Aged , Natriuresis/drug effects , Randomized Controlled Trials as Topic/statistics & numerical data , Tetrazoles/blood , Tetrazoles/urine , Valsartan , Young AdultABSTRACT
A simple method, incorporating protein-precipitation/organic backwashing and liquid chromatography-tandem mass spectrometry (LC-MS-MS), has been successfully developed for the simultaneous analysis of four highly water-soluble and less volatile herbicides (paraquat, diquat, glufosinate and glyphosate) in ante- and postmortem blood, urine and gastric content samples. Respective isotopically labeled analogs of these analytes were adopted as internal standards. Acetonitrile and dichloromethane were used for protein precipitation and organic solvent backwashing, respectively, followed by injecting the upper aqueous phase into the LC-MS-MS system. Chromatographic separation was achieved using an Agilent Zorbax SB-Aq analytical column, with gradient elution of 15 mM heptafluorobutyric acid and acetonitrile. Mass spectrometric analysis was performed under electrospray ionization in positive-ion multiple reaction monitoring mode. The precursor ions and the two transition ions (m/z) adopted for each of these four analytes were paraquat (185; 169 and 115), diquat (183; 157 and 78), glufosinate (182; 136 and 119) and glyphosate (170; 88 and 60), respectively. Analyte-free blood and urine samples, fortified with the analytes of interest, were used for method development/validation and yielded acceptable recoveries of the analytes; interday and intraday precision and accuracy data; calibration linearity and limits of detection and quantitation. This method was successfully incorporated into an overall analytical scheme, designed for the analysis of a broad range of compounds present in postmortem samples, helpful to medical examiners' efforts to determine victims' causes of death.
Subject(s)
Herbicides/metabolism , Aminobutyrates/blood , Aminobutyrates/metabolism , Aminobutyrates/urine , Autopsy , Chromatography, Liquid , Coroners and Medical Examiners , Death , Diquat/blood , Diquat/metabolism , Diquat/urine , Forensic Toxicology , Glycine/analogs & derivatives , Glycine/blood , Glycine/metabolism , Glycine/urine , Herbicides/blood , Herbicides/urine , Paraquat/blood , Paraquat/metabolism , Paraquat/urine , Tandem Mass Spectrometry , GlyphosateABSTRACT
Population level variation of drug metabolism phenotype (DMP) has great implications in treatment outcome, drug-related side effects, and resistance development. In this study, we used a gas chromatography-time of flight-mass spectrometry (GC-TOF-MS)-based untargeted urine metabolomics approach to understand the DMP of a tuberculosis (TB) patient cohort (n= 20) from Tripura, a state in the northeastern part of India. Urine samples collected at different postdose time points (2 h, 6 h, 12 h, 24 h, 36 h, and 48 h) from these newly diagnosed TB patients receiving first-line anti-TB drugs were analyzed, and we have successfully detected three of the four first-line drugs,viz, isoniazid (INH), ethambutol (ETB), and pyrazinamide (PZA). The majority of their known metabolites, acetyl-isoniazid (AcINH), isonicotinic acid (INA), isonicotinuric acid (INTA), 2,2'-(ethylenediimino)-dibutyric acid (EDBA), 5-hydroxypyrazinamide (5OH-PZA), pyrazinoic acid (POA), and 5-hydroxypyrazinoic acid (5OH-POA), were also detected. Analyzing the variation in abundances of drugs and their known metabolites and calculating the metabolic ratios in these samples, we offer comprehensive DMP information on this small patient cohort that represents Tripura, India. The majority (75%) of these patients are found to be slow acetylators of INH. The average metabolic ratios of POA/PZA and 5OH-POA/POA are 3.16 ± 3.03 and 6.09 ± 6.15, respectively. Employing correlation analysis of the metabolomics metadata and a manual prediction of drug catabolism, we have proposed 2-aminobutyric acid (AABA) as a novel metabolite of ETB. These observations indicate the usefulness of GC-MS-based metabolomics to characterize the DMP at a population level and also to identify novel drug metabolites.
Subject(s)
Aminobutyrates/urine , Antitubercular Agents/urine , Ethambutol/urine , Metabolomics , Tuberculosis, Pulmonary/urine , Adult , Aged , Aged, 80 and over , Antitubercular Agents/therapeutic use , Biotransformation , Case-Control Studies , Chromatography, Gas , Ethambutol/therapeutic use , Female , Humans , India , Isoniazid/therapeutic use , Isoniazid/urine , Male , Middle Aged , Mycobacterium tuberculosis/physiology , Pyrazinamide/therapeutic use , Pyrazinamide/urine , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tuberculosis, Pulmonary/drug therapy , Tuberculosis, Pulmonary/microbiologyABSTRACT
A rapid, specific, and sensitive method for the simultaneous quantitation of organophosphates (fenitrothion (MEP), malathion, and phenthoate (PAP)), glufosinate (GLUF), and glyphosate (GLYP) in human serum and urine by gas chromatography-mass spectrometry (GC-MS) has been validated. All of the targeted compounds together with the internal standard were extracted from the serum and urine using a mix-mode TiO-C(18) monolithic spin column. The recovery of organophosphates from serum and urine ranged from 12.7 to 49.5%. The recovery of GLUF and GLYP from serum and urine ranged from 1.9 to 7.9%. The intra- and inter-accuracy and precision (expressed as relative standard deviation, %RSD) were within 96.7-107.7% and 4.0-13.8%, respectively. The detection and quantitation limits for serum and urine were 0.1 and 0.1 µg/ml, respectively, for organophosphates, 0.1 and 0.5 µg/ml, respectively for GLUF and GLYP. The method had linear calibration curves ranging from 0.1 to 25.0 µg/ml for organophosphates and 0.5-100.0 µg/ml for GLUF, and GLYP. The validated method was successfully applied to a clinical GLYP poisoning case.
Subject(s)
Aminobutyrates/blood , Aminobutyrates/urine , Glycine/analogs & derivatives , Organophosphorus Compounds/blood , Organophosphorus Compounds/urine , Gas Chromatography-Mass Spectrometry , Glycine/blood , Glycine/urine , Humans , Polymethacrylic Acids/chemistry , Sensitivity and Specificity , Titanium/chemistry , GlyphosateABSTRACT
This paper describes an assay for the determination of glyphosate (GLYP), glyphosate metabolites [(aminomethyl) phosphonic acid] (AMPA), and glufosinate (GLUF) in human serum. After protein precipitation using acetonitrile and solid-phase extraction, serum samples were derivatized and analyzed by gas chromatography-mass spectrometry (GC-MS). The assay was linear over a concentration range of 3-100.0 microg/ml for GLYP, AMPA, and GLUF. The overall recoveries for the three compounds were >73%. The intra- and inter-day variations were <15%. Precision and accuracy were 6.4-10.6% and 88.2-103.7%, respectively. The validated method was applied to quantify the GLYP and AMPA content in the serum of a GLYP-poisoned patient. In conclusion, the method was successfully applied for the determination of GLYP and its metabolite AMPA in serum obtained from patient of GLYP-poisoning.
Subject(s)
Aminobutyrates/blood , Gas Chromatography-Mass Spectrometry/methods , Glycine/analogs & derivatives , Herbicides/blood , Organophosphonates/blood , Aminobutyrates/poisoning , Aminobutyrates/urine , Drug Stability , Glycine/blood , Glycine/poisoning , Glycine/urine , Herbicides/poisoning , Herbicides/urine , Humans , Isoxazoles , Linear Models , Male , Middle Aged , Organophosphonates/urine , Reference Standards , Reproducibility of Results , Sensitivity and Specificity , Suicide, Attempted , Tetrazoles , GlyphosateABSTRACT
Stimulants are banned by the World Anti-Doping Agency (WADA) if used "in competition". Being the analysis of stimulants presently carried out on urine samples only, it might be useful, for a better interpretation of analytical data, to discriminate between an early intake of the substance and an administration specifically aimed to improve the sport performance. The purpose of the study was to investigate the differences, in terms of excretion/disappearance of drugs, between urine and oral fluid, a sample that can reflect plasmatic concentrations. Oral fluid and urine samples were collected following oral administration of the following stimulants: modafinil (100 mg), selegiline (10 mg), crotetamide/cropropamide (50 mg each), pentetrazol (100 mg), ephedrine (12 mg), sibutramine (10 mg), mate de coca (a dose containing about 3mg of cocaine); analysis of drugs/metabolites was carried out by gas chromatography/mass spectrometry (GC/MS) in both body fluids. Our results show that both the absolute concentrations and their variation as a function of time, in urine and in oral fluid, are generally markedly different, being the drugs eliminated from urine much more slowly than from oral fluid. Our results also suggest that the analysis of oral fluid could be used to successfully complement the data obtained from urine for "in competition" anti-doping tests; in all those cases in which the metabolite(s) concentration of a substance in urine is very low and the parent compound is not detected, it is indeed impossible, relying on urinary data only, to discriminate between recent administrations of small doses and remote administrations of higher doses.
Subject(s)
Central Nervous System Stimulants/urine , Doping in Sports , Saliva/chemistry , Substance Abuse Detection/methods , Adult , Aminobutyrates/analysis , Aminobutyrates/pharmacokinetics , Aminobutyrates/urine , Benzhydryl Compounds/analysis , Benzhydryl Compounds/pharmacokinetics , Benzhydryl Compounds/urine , Calibration , Central Nervous System Stimulants/analysis , Central Nervous System Stimulants/pharmacokinetics , Cocaine/analysis , Cocaine/pharmacokinetics , Cocaine/urine , Crotonates/analysis , Crotonates/pharmacokinetics , Crotonates/urine , Ephedrine/analysis , Ephedrine/pharmacokinetics , Ephedrine/urine , Female , Gas Chromatography-Mass Spectrometry/methods , Humans , Modafinil , Selegiline/analysis , Selegiline/pharmacokinetics , Selegiline/urine , Time FactorsABSTRACT
We have established a new HPLC method for derivatizing and quantifying glufosinate (GLUF) in human serum and urine using p-nitrobenzoyl chloride (PNBC). The p-nitrobenzoyl derivative of GLUF (PNB-GLUF) was produced quantitatively over 10 min at room temperature. PNB-GLUF possesses the property of ultraviolet (UV) light absorption with a lambda(max) of 272.8 nm, and was isolated from biological specimens by reversed-phase chromatography using Inertsil Ph-3. In experiments at a UV wavelength of 273 nm, GLUF has a quantitative detection limit of 0.005 microg/ml, and when it was added to both serum and urine to yield concentrations of 0.1-1000 microg/ml, its recovery rate was quite satisfactory: at least 93.8% in all cases. Further, the measured amounts of GLUF in 23 serum samples from patients intoxicated by ingestion of GLUF compared favorably with those obtained by fluorescence derivatization-HPLC using 9-fluorenylmethyl chloroformate (R=0.998). This technique of analysis is, in addition, applicable for Glyphosat, which possesses a chemical structure resembling that of GLUF, and it will be of great use in the determination of these two compounds.
Subject(s)
Aminobutyrates/analysis , Chromatography, High Pressure Liquid/methods , Spectrophotometry, Ultraviolet/methods , Aminobutyrates/blood , Aminobutyrates/urine , Humans , Sensitivity and SpecificityABSTRACT
Incidents of poisoning in humans caused by the ingestion of the glufosinate ammonium containing herbicides are gradually increasing in Japan. This poisoning is characterized by various neurological symptoms such as disturbances of consciousness, convulsions and apnea which appear after an asymptomatic interval of several hours. We studied the toxicokinetics of glufosinate in a patient with this poisoning successfully treated without extracorporeal hemopurification. A 65-year-old male ingested BASTA, which contains 20% w/v of glufosinate ammonium, about 300 ml, more than the estimated human toxic dose. Four and a half hours after ingestion, he showed speech ataxia and systemic tremor. He was prophylactically intubated before the occurrence of serious respiratory failure. After 5 days of artificial ventilation he was extubated and discharged without any sequelae. We studied the serial change of serum glufosinate concentration every 3-6 h and assessed the urinary excretion of glufosinate every 24 h. The absorbed amount of glufosinate was estimated from the cumulative excreted in urine. Toxicokinetic analysis was performed using the two-compartment model. The changes in serum glufosinate concentration exhibited T1/2alpha of 1.84 and T1/2beta of 9.59 h. The apparent distribution volume at beta-phase and the total body clearance were 1.44 l/kg and 86.6 ml/min, respectively. Renal clearance was estimated to be 77.9 ml/min. The indication for extracorporeal hemopurification for this poisoning has been discussed.
Subject(s)
Aminobutyrates/poisoning , Herbicides/poisoning , Aged , Aminobutyrates/blood , Aminobutyrates/pharmacokinetics , Aminobutyrates/urine , Ataxia/chemically induced , Ataxia/metabolism , Herbicides/blood , Herbicides/pharmacokinetics , Herbicides/urine , Humans , Male , Metabolic Clearance Rate , Tremor/chemically induced , Tremor/metabolismABSTRACT
The respiratory stimulant prethcamide is a mixture of equal parts of crotethamide and cropropamide. A specific and sensitive gas chromatographic method for the determination of crotethamide and cropropamide in horse plasma and urine is described. Both components of prethcamide were extracted from plasma and urine into dichloromethane. The extracts were analyzed by capillary gas chromatography with thermionic detection in the nitrogen-specific detection mode. The lower limits of quantitation were 4.0 ng ml-1 of plasma and 10.0 ng ml-1 of urine. Calibration curves were linear from 2.0-100 ng ml-1 of plasma for both components. Pharmacokinetic parameters for crotethamide and cropropamide after intravenous and oral dosing were estimated by analysis of plasma concentration versus time data. The total plasma clearance of cropropamide was greater than that of crotethamide and both values were greater than 5 ml min-1 kg-1. Renal clearance values of the two drugs were comparable and were much less than estimates of filtration clearance values in horses, indicating extensive re-absorption of both components from the renal tubules. Both compounds were metabolized by N-demethylation of the [(dimethylamino)-carbonyl]-propyl moiety and these metabolites were excreted in urine. The method was demonstrated to be suitable for detecting illicit administration of prethcamide to competition horses.
Subject(s)
Aminobutyrates/pharmacokinetics , Horses/metabolism , Administration, Oral , Aminobutyrates/blood , Aminobutyrates/urine , Animals , Blood Proteins/metabolism , Chromatography, Gas , Crotonates/blood , Crotonates/pharmacokinetics , Crotonates/urine , Female , Horses/blood , Horses/urine , Injections, Intravenous , Protein BindingSubject(s)
Aminobutyrates/analysis , Herbicides/analysis , Aminobutyrates/blood , Aminobutyrates/urine , Chromatography, Paper , Gastric Juice/analysis , Glycine/analogs & derivatives , Glycine/analysis , Glycine/blood , Glycine/urine , Herbicides/blood , Herbicides/urine , Humans , Organophosphorus Compounds/analysis , Organophosphorus Compounds/blood , Organophosphorus Compounds/urine , GlyphosateABSTRACT
Urine from untreated patients with various tumours and controls has been examined for the excretion of beta-aminoisobutyric acid and uric acid. The patients were classified into four groups: I, beta-aminoisobutyric acid and uric acid both normal; II, beta-aminoisobutyric acid normal, uric acid elevated; III, beta-aminoisobutyric acid elevated, uric acid normal; IV, beta-aminoisobutyric acid and uric acid both elevated. Uric acid was used as an indicator for tissue-breakdown. Pseudouridine being a specific parameter for t-RNA degradation was estimated for comparison. Increased urinary concentrations of beta-aminoisobutyric acid were frequently found in tumour patients, especially in patients with leukaemia and non-Hodgkin lymphoma. Tissue breakdown being the cause of the beta-aminoisobutyric aciduria could only be considered in part of the patients. Moreover, strongly elevated ratios of beta-aminoisobutyric acid to uric acid were found. Urinary patterns of pyrimidines and purines were determined in order to exclude other abnormalities. The results are discussed in relation to thymine metabolism and renal function.
Subject(s)
Aminobutyrates/urine , Neoplasms/urine , Thymine/urine , Adolescent , Child , Child, Preschool , Humans , Infant , Leukocyte Count , Purines/urine , Pyrimidines/urine , Uric Acid/urineABSTRACT
Crotethamide and cropropamide, both components of the respiratory stimulant prethcamide, are metabolized in humans by demethylation of the [(dimethylamino)-carbonyl]-propyl moiety. The resulting metabolites are characterized by gas chromatography-mass spectrometry of urinary extracts. The use of HCl to prevent losses by volatilization during the evaporation step, combined with methanol as solvent, complicates gas liquid chromatographic analysis of prethcamide. The resulting artifacts are identified.
Subject(s)
Aminobutyrates/metabolism , Administration, Oral , Aminobutyrates/administration & dosage , Aminobutyrates/urine , Gas Chromatography-Mass Spectrometry/methods , Humans , IsomerismABSTRACT
A new amino acid has been isolated from the normal human urine. The chemical structure of the amino acid was determined to be alpha-hydroxy-beta-keto-gamma-aminobutyric acid based on its physical properties involving NMR, infrared and mass spectra, as well as chemical degradation and synthesis. In six healthy adults the urinary contents of the new amino acid were 3.2--4.5 mumol/24 h.
Subject(s)
Aminobutyrates/urine , gamma-Aminobutyric Acid/urine , Chromatography, Gas , Humans , Magnetic Resonance Spectroscopy , Mass Spectrometry , Periodic Acid , gamma-Aminobutyric Acid/analogs & derivatives , gamma-Aminobutyric Acid/chemical synthesisABSTRACT
Initial urinary catecholamine metabolite and amino acid excretion patterns were examined in 54 children with neuroblastoma. The relationships between prognosis and age at diagnosis, stage of disease, primary site, and histologic grade of tumor were similar in this population to those found in previous studies, but only age and stage were found to be independent prognostic variables. Prognosis in disseminated disease was found to correlate directly with the urinary vanilmandelic acid (VMA)/homovanillic acid (HVA) ratio but not with the absolute levels of HVA. The presence of the dopa metabolite, vanillactic acid, as well as increased amounts of cystathionine and/or low levels of VMA indicated poor prognosis. These results are consistent with the hypothesis that biochemically primitive neuroblastomas deficient in dopamine beta-hydroxylase are move virulent than their mature analogues which produce epinephrine, norepinephrine, and their metabolites.
Subject(s)
Catecholamines/urine , Neuroblastoma/urine , Adrenal Gland Neoplasms/mortality , Adrenal Gland Neoplasms/pathology , Adrenal Gland Neoplasms/urine , Aminobutyrates/urine , Child, Preschool , Cystathionine/urine , Female , Homovanillic Acid/urine , Humans , Infant , Infant, Newborn , Male , Neuroblastoma/mortality , Neuroblastoma/pathology , Prognosis , Vanilmandelic Acid/urineABSTRACT
Under the conditions of disynchronization by the manipulation of both the alternation of light and darkness and the availability and unavailability of food, circadian rhythms characterize the excretion of several amino acids by inbred LOU rats bearing an immunocytoma. Large amplitude rhythms can be demonstrated for urinary beta-aminoisobutyric acid, beta-alanine, phenylalanine and tyrosine. Under the same conditions of disynchronization, control animals excrete the same compounds also with a marked circadian variation but at an invariably lower average rate. These excretory rhythms, along with those demonstrated earlier for polyamines and light-chains, are of interest as potential markers for the chronotherapy of cancer.
Subject(s)
Amino Acids/urine , Circadian Rhythm , Lymphoma/urine , Aminobutyrates/urine , Animals , Diet , Light , Neoplasms, Experimental/urine , Phenylalanine/urine , Rats , Rats, Inbred Strains , Tyrosine/urine , beta-Alanine/urineABSTRACT
A study of amino acids determined by sequential Multi-sample Amino Acid Automatic Analyzer in plasma, urine and cerebrospinal fluid (CSF) in patients with Friedreich's ataxia and control subjects has revealed a number of mathematically significant variations from normal. Of practical physiological importance are the following: a high urinary excretion of alanine with slightly elevated plasma levels; a low plasma and CSF concentration of aspartic acid in the presence of normal urinary values and finally a low CSF concentration of taurine accompanied by normal plasma levels, but elevated urinary output and renal clearance rates. We postulate that the modifications in alanine and aspartic acid are less specific and probably secondary, but there could be a genetic defect in the membrane transport of taurine and the other beta-amino acids in Friedreich's ataxia.
Subject(s)
Amino Acids/metabolism , Friedreich Ataxia/metabolism , Alanine/urine , Aminobutyrates/blood , Aminobutyrates/urine , Asparagine/blood , Aspartic Acid/blood , Aspartic Acid/cerebrospinal fluid , Glycine/blood , Histidine/blood , Humans , Isoleucine/urine , Ornithine/blood , Phenylalanine/blood , Phosphatidylethanolamines/urine , Sarcosine/urine , Serine/urine , Taurine/cerebrospinal fluid , Taurine/urine , Tyrosine/urine , Valine/urineABSTRACT
The effects of inhibitors of diamine oxidase (EC 1.4.3.6), monoamine oxidase (EC 1.4.3.4) and 4-aminobutyrate aminotransferase (EC 2.6.1.19) on the catabolism of putrescine in mice in vivo were studied. Diamine oxidase inhibitors and carboxymethoxylamine (amino-oxyacetate) markedly inhibit the metabolism of [(14)C]putrescine to (14)CO(2), but affect different enzymes. Aminoguanidine specifically inhibits the mitochondrial and non-mitochondrial diamine oxidases, whereas carboxymethoxylamine specifically inhibits 4-aminobutyrate transamination by the mitochondrial pathway. Hydrazine inhibits at both sites, and results in increased concentrations of 4-aminobutyrate in brain and liver. Pretreatment of mice with carboxymethoxylamine and [(14)C]putrescine leads to the urinary excretion of amino[(14)C]butyrate. Carboxymethoxylamine does not affect the non-mitochondrial pathway of putrescine catabolism, as the product of oxidative deamination of putrescine in the extramitochondrial compartment is not further oxidized but is excreted in the urine as derivatives of 4-aminobutyraldehyde. Another catabolic pathway of putrescine involves monoamine oxidase, and the monoamine oxidase inhibitor, pargyline, decreases the metabolism of [(14)C]putrescine to (14)CO(2)in vivo. Catabolism of putrescine to CO(2)in vivo occurs along different pathways, both of which have 4-aminobutyrate as a common intermediate, in contrast with the non-mitochondrial catabolism of putrescine, which terminates in the excretion of 4-aminobutyraldehyde derivatives. The significance of the different pathways is discussed.