[Simultaneous detection methods for abused diuretics in urine and a quantitative determination method for trichlormethiazide in urine].
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Biomedical subjects
Publications and source records attributed to Z Tamura.
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A method for the determination of catecholamine sulfoconjugate isomers (CA-S) in urine was developed. The photo-induced fluorogenic reaction of CA-S with ethylenediamine reported previously was applied to the postcolumn labeling of HPLC for sensitive and selective detection. Special equipment for the reaction was made with a uv-irradiation lamp and a reaction coil of Teflon tubing inside a temperature-controlled reaction box. Lower determination limits of this system were 1 to 2 pmol. Urine samples pretreated with small ion-exchange resin columns were subjected to HPLC. Peaks corresponding to CA-S were identified quantitatively by two different separation methods. Thus, all six CA-S were first detected in the urine of normal individuals. The excretion rates of dopamine 3-sulfate, dopamine 4-sulfate, norepinephrine 3-sulfate, norepinephrine 4-sulfate, epinephrine 3-sulfate, and epinephrine 4-sulfate were 420 +/- 240, 98 +/- 55, 86 +/- 95, 15 +/- 14, 18 +/- 7, and 3 +/- 1 ng/min (+/- SD), respectively (n = 5).
[2H2]-dopamine-3-0-sulfate (DM-3-0-S) and [2H2]-dopamine-4-0-sulfate (DM-4-0-S) were synthesized to investigate the possibility of their being substrates for catechol-0-methyltransferase (COMT). [2H5]-3-0-methyldopamine (3-0-Me-DM) and [2H5]-4-0-methyldopamine (4-0-Me-DM) were also synthesized as internal standards for the determination of enzymatic products by gas chromatography-mass spectrometry (GC-MS). [2H2]-DM-3-0-S or [2H2]-DM-4-0-S was incubated at 37 degrees for 60 min in the presence of S-adenosyl-L-methionine with a crude enzyme preparation obtained from rat liver homogenate. The incubation mixture was treated with 0.5N HCl at 100 degrees C for 1h to hydrolyze the remaining sulfate moiety. The reaction products were extracted with an Amberlite XAD-4, derivatized with pentafluoropropionic anhydride and determined by GC-MS. When [2H2]-DM-3-0-S was used as a substrate, [2H2]-3-0-Me-DM was found to be a major product accompanied by [2H2]-4-0-Me-DM as a minor product. The ratio of [2H2]-3-0-Me-DM to [2H2]-4-0-Me-DM was found to be 26:1, while the ratio was 5.4:1 when [2H2]-dopamine was used as a substrate. When [2H2]-DM-4-0-S served as a substrate, [2H2]-3-0-Me-DM was preferentially produced without detectable formation of [2H2]-4-0-Me-DM.
The extent and rate of absorption of phenytoin (PHT) from tablet and powder were studied in four healthy adult volunteers. It was demonstrated by urinary and fecal excretion that the almost all quantity of PHT in tablet was absorbed through the gastrointestinal tract, and the observed values of the estimated free concentration (Cest.f) estimated from mixed saliva concentration of PHT in the multiple dose were in fair agreement with the calculated values of that by using computer simulation in case of tablet. On the contrary, the variations were observed in Cest.f using therapeutic dose of PHT powder. The values of Cest.f at steady-state in tablet administration were higher than those in powder administration. The absorption ratio of PHT powder was low and variable, and decreased upon increase of dose. The ratio calculated from the Cest.f values of both dosage forms at steady-state were in good correspondence to the observed values of PHT excreted in feces.
The dissolution profiles of furosemide in various solutions were studied with plain tablet and retard capsule of furosemide. The rate constant and percent dissolution in retard capsule were lower than that in plain tablet. Clinical pharmacokinetics and diuretic effect of furosemide after oral administration of two dosage forms were also studied with 3 normal subjects and 3 cirrhotic patients. In normal subjects, the extent of furosemide absorption from retard capsule was 45% of plain tablet. The daily urine volume after oral administration of two dosage forms was comparable, however, quite different profiles were observed between these dosage forms. In patients, the extent of furosemide absorption in retard capsule was one half that of plain tablet. The dose of retard capsule was increased without adverse reactions, thus decreasing of ascites, which was not observed with plain tablet, was achieved with increasing urine volume.
A 20% phenytoin (PHT) plain mixture with excipients (20%PM) and three PHT products prepared by wet granulation, which are 20% fine granule (20%FG), 99% air-dried fine granule (99%FG-A) and 99% freeze-dried fine granule (99%FG-F), were prepared. The extents of PHT absorption from these products and the Aleviatin Fine Granules (97%FG) prepared with microcrystalline PHT powder were compared with those from commercially available PHT powder (Aleviatin) and tablet (Hydantol Tablet 25 mg), which are of the Pharmacopoeia of Japan grade, in healthy adult volunteers. In single dose study, the extents of PHT absorption from the powder, 20%PM, 20FG, 99%FG-A, 99%FG-F, 97%FG and tablet were 89.7, 92.2, 99.0, 96.7, 99.1, 99.1 and 99.3%, respectively. The property of almost complete absorption of PHT from the product was shown in the 20%FG, 99%FG-F and 97%FG similar to the tablet. In multiple dose study, the minimum and the average estimated free concentrations of PHT at steady-state for 99%FG-F and 97%FG were nearly equal to those for the tablet, and were higher than those for the powder. In epileptic patients, the plasma PHT concentrations were increased when dosage form was changed from the powder to 99%FG-F. However, the plasma PHT concentrations were scarcely altered when dosage form was changed from the tablet to 99%FG-F. The change in dosage forms from the tablet to 99%FG-F and 97%FG or opposite direction can be done without causing toxicity in epileptic patients, so long as these products are used at the same amounts as PHT.
The investigation was undertaken to study the neurological symptoms in rats caused by maintaining high plasma concentration of about 30 nmol/ml or more, of clioquinol. Clioquinol suspension which was prepared using polysorbate 80 was administered intraperitoneally to rats and plasma and tissue concentrations were determined. On administration of clioquinol of 100 and 200 mg/kg, the mean plasma concentrations of clioquinol reached maximum values of 30 and 58 nmol/ml, respectively, after 0.5-1 h and thereafter decreased rapidly. With 400 mg/kg, however, plasma concentration reached maximum value of about 75 nmol/ml and fell slowly. By single and repeated administration of the suspension, clioquinol was distributed in the liver and kidney at a high concentration, and also in the nervous system. In experiments on appearance of neurotoxicity in rats by repeated administration of the suspension, all of 10 rats administered intraperitoneally with 100 mg/kg/d did not develop any neurological symptoms for about 30 d. On the other hand, one of 10 and 7 of 13 rats administered with 200 and 400 mg/kg/d, respectively, developed ataxia in the hind legs or all legs on the 3rd to the 12th day after starting administration. Pathologically, a slight change of the peripheral nerve, central chromatolysis of the anterior horn neuron and severe neuronal degeneration of the Ammon's horn were observed in the rats with ataxia.
Plasma concentrations of clioquinol and its metabolites after single or repeated oral administration of clioquinol, absorption region of clioquinol in gastrointestinal tract, and intestinal metabolism were studied in rats. Plasma concentrations of clioquinol after oral administration of four different doses (20, 100, 200 and 400 mg/kg) were lower than those of the two metabolites, clioquinol glucuronide and sulfate. Mean maximal plasma concentration of unchanged drug was in the range of 1-8 nmol/ml. Clioquinol was absorbed poorly from the stomach and fairly from the small intestine. Bile was an important route for excretion of clioquinol in rats. The mesenteric venous plasma from the closed intestinal loops of both jejunal and ileal regions was analyzed for clioquinol and the metabolites and it was found that clioquinol glucuronide was formed predominantly in both regions. From the results of the present studies, intestinal metabolism of clioquinol can be pointed out as a major factor for difficulty to cause clioquinol intoxication.
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Explore the source record for details and available documents.
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A method for the determination of biogenic 5-hydroxy- and 5-methoxyindoles using high-performance liquid chromatography with fluorescence detection was developed. The specific fluorescence of these indoles induced by perchloric acid was utilized as the post-column detection system. The determination limits were 3 pmol for 5-hydroxy-L-tryptophan and 5 pmol for serotonin (5-HT) and 5-hydroxyindole-3-acetic acid (HIAA). The method was applied to the determination of 5-HT and HIAA in rat brain tissues using 5-hydroxygramine as the internal standard.
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Spinal reflex and neuromuscular functions were investigated in the beagles ingesting clioquinol over a long period. The twitch tension, the refractory period, the summation of contraction and the discharge of the muscle spindles of the gastrocnemius muscle were not significantly damaged in the treated beagles, whereas the response with long latency of the spinal reflex disappeared completely during the administration not in the control but in the treated dogs. The meanings of disappearance of the spinal reflex in the treated dogs are discussed in relation to the degenerative change in the fasciculus gracilis at the cervical level of the spinal cord.
The parallel flow dialysis technique was improved for application to the detection of drug-binding proteins in a column chromatographic effluent. To prevent the baseline drift, the pressures of both protein and drug channels were maintained equal during chromatography, and Brij-35 was added to the solvents. The improved method was successfully applied to the detection of methyl orange-binding proteins in human serum and bromphenol blue-binding proteins in rat liver homogenate.
A parallel-flow dialysis technique utilizing a Technicon dialyzer and a constant-flow system has been described for the detection of drug-binding proteins. The effect of temperature, flow-rate and drug concentration was investigated by measuring the efficiency of dialysis and detecting the binding of methyl orange to bovine serum albumin. The larger response was shown to be achieved by increasing the efficiency of dialysis or the drug concentration. The present method will enable the continuous monitoring of drug-binding proteins.
Binding of biotin to resting cells of Bifidobacterium breve N4, which grew in a biotin-deficient medium, was independent of pH from 1 to 9 and of temperature below 50 C. It was not inhibited by metabolic inhibitors including sulfhydryl reagents, but it was inhibited by treatment with 80% ethanol or 5% trichloroacetic acid. It was also competitively inhibited by biotin-sulfone, but not by tetrahydrothiophene nor dethiobiotin. The binding constant was calculated to be 3.3 X 10(8) M--1. The amount of biotin unextractable with hot water, representing part of the transported biotin, increased gradually for 20 min, this increase was inhibited by NaF, hydroxylamine and low temperature. 14C-biotin on the cells was displaced by cold biotin and biotin-sulfone; the displacement was not inhibited by metabolic inhibitors, but it was dependent on temperature. A few minutes after binding, the biotin was released to the medium. The release was dependent on pH and temperature, was affected by energy sources and was inhibited by metabolic inhibitors, e.g. NaF, p-chloromercuribenzoic acid and hydroxylamine. It could be stopped at any time by cooling to 0 C or by adding NaF, and the amount of accumulated biotin did not increase under those conditions. These results suggest that the binding sites on the cell surface decreased in number or in their binding affinity for biotin through an energy-dependent process.