A method for the simultaneous determination of chlorpromazine (CPZ) and levomepromazine (LPZ) in human urine has been developed using high-performance liquid chromatography (HPLC) with electrochemical detectlon. CPZ and LPZ were separated by a reversed phase HPLC on μ Bondapak C18 (4.6mm i.d. ×250mm) using mobile phase of methanol-ethanol-tetrahydrofuran-acetate buffer (pH 5.0)(53.5: 9.0: 2.5: 35.0, v/v), containing 20mM NaClO4, and the separation was completed within 10min.Minimum detectable amounts on this detector were 500Pg for CPZ and 200Pg for LPZ with higher sensitivity than UV detector.Recoveries of CPZ and LPZ (2 and 20μg) from urine were 84.0% and 98.0%, respectively. This method was apPlied to the determination of CPZ and LPZ concentrations in the urine of schizophrenics receiving simultaneous dose of CPZ and LPZ.
The actnon of androgen hormones has been estimated by morphological or behavior changes. In this study, the androgenic action of an ether extracts of musk from the gland secretions of prepucial follicler in male musk deer (Moschus moschiferus Linné) was examined. ln partncular the effect of fraction Vll was estimated by assesing electrophoretic changer both in such submaxillary gland enzymes as arginine peptndase and glucose-6-phosphate dehydrogenase (G6PD), which show sexual dimor phism in zymogram of glandular extracts. Arginine peptidase actnvnty in male mice is markedly higher than that in female mlce, but G6PD level does not exhibit a clear-cut difference between the male and the female.In the male, arginine peptidase activity as well as G6PD activity were converted itto the female pattern by castration.When testosterone was adminrtrated to castrated mice, argine peptidase began to increase gradually between day 2 and day 6, and then decreased after day 6.But G6PD activity showed a decrease after day 3 with increaring testosteronedosage. Moreover.the D band of G6PD was not affected by testosterone treatment.The G band of arginine peptidase in zymograms seems to be reprerent female specnfic activity. An action similar to that of androgen hormones on arginine peptidase and G6PD, was revealed in ether extracts of musk and its fraction Vll by electrophoretic analysis of submaxillary gland enzymes such as arginine peptidase and G6PD.From this result, we can confirm that ether extracts of musk and its fraction Vll contained the androgen-like substance. Electrophoretic changes in arginine peptndase and G6PD may be employed to estimate the androgenic action of certain kinds of extractrs such as those found in Ornental drugs.
A simple fluorometric method for the estimation of urinary pregnanednol (P2) was devised.The urine sample (non-pregnancy or early pregnancy urine: 1.0ml, middle or late pregnancy urine: 0.2-0.5ml) was hydrolyzed with β-glucuronidase and extracted with methylene chloride.Urinary ketosteroids were eliminated by the Girard reaction and estrogens were eliminated by washing the methylene chloride extract with 5N-NaOH.The extract was evaporated, treated with sulfuric acid at 100° for 10 minutes, and measured by fluorometry at 485nm excitation wavelength and 515nm emission wavelength. The precision (c.v.) of the method was 6.7% (range: 1.49-172μg/ml, mean: 1.64μg/ml) for the urine specimen of a female in follicular phase (n=10) or 6.31% for 5μg/ml P2 standard solution (n=10).The recovery of P2 added to urine was 96.0% (mean of ten repeated experiments.range: 91.6-102.0%).The detectable limit was 0.2μg per tube or 1μg/ml urie.Some examples of the P2 assay in normal menstrual cycle and in pregnancy have been reported.
The stick methods, utilizig Sodium Nitroprusside, are generally used for the monitoring of ketonuria in clinical laboratory tests.The influence of Cephacetrile and various other Cephem series of antibiotics on this test was studied. Cephacetrile iterfered with the ketonuria test to produce reddish orange color differig from the purple Color of ketoacidosis when ketone body's absent in the urine. Other Cephems almost always gave a dark orange color with the passage of time in the same way. This kind of coloration may lead to incorrect readigs of ketonuria when the test is applied to urine sample containing Cephems or its degradation, metabolites. Fresh urine should therefore be used in the test.Other countermeasures are needed when testing urie in patients receiving Cephacetrile and in cases where fresh urine samples are difficult to obtain and when it is difficult to read differences in the shades of dark orange due to the inexperience of the tester. ln present study, we established a simple and useful method against this iterference. The method involves the acidification of the urie sample with N HCl and treatment with a small activated charcoal column, and war tried in the determination of ketone bodies in urine obtaied from healthy volunteers and in patients receiving Cephems. Satisfactory rerults without any iterference were obtained.
p-Guanidinobenzoate derivatives were prepared, and their inhibitory effects on trypsin, plasmin, pancreatic kallikrein, plasma kallikrein, thrombin, Clr and Clesterase were examined.Among the various inhibitors tested, 6-amidino-2-naphthyl-4-guanidino benzoate·dihydrochrolide, 6-(β-amidinoethenyl) phenyl-4-guanidinobenzoate·dimethanesul.fonate and 6-amidino-2-benzoylphenyl-4-guanidin- benzoate·dimethanesulfonate were the moste ffective inhibitors of trypsin, plasmin, pancreatic kallikrein, plasma kallikrein and thrombin, and they strongly inhibited the esterolytic activities of Clr and Clesterase, and then strongly inhibited complemernt-mediated hemolysis.
lndolacetic acid is one of the metabolites of tryptophan, As its levels in body fluid is very Iow, it has been difficult to estimate it by conventional analytical methods. In the present study, an attempt was made to employ gas Chromatogram/mass spectrometry (GC/MS) for the determination of the Ievels in serum from patients with several disease, such as gastric and cobn cancer, hepatoma and other gastroenteric diseasee. For GC/MS was used Finnigan Quardrupole Type Mass Spectrometer. Quantitative analyses of indolaceticacid in serum were performed by the multiple ion detectation technique, because its sensitivity was higher than the total ion monitoring technique, and indolcarbomc acid was used as an internal standard, which is absent in natural organisms. Possible usefulness for diagnostic examination of cancers was also discussed.
Enzymatic determination of urinary estriol in pregnancy using thin-layer chromatographic separation has been described. The enzymatic assay of estriol is performed with peroxidase as a catalyzer, hydrogen peroxide as a hydrogen acceptor and estriol as a substrate of hydrogen donor where hydrogen peroxide is consumed depending on the concentration of estriol, and remains of hydrogen peroxide is determined colorimetrically using 4-aminoantipyrine, phenol, and peroxidase. Urinary estriol purified and separated by thinlayer chromatography is determined enzymatically. The correlation obtained by other colorimetric method and this enzymatic method is fairly good.
we established a new procedure for the separation and quantitation of amino acids in human plasma and cerebrospinal fluid using high performance liquid chromatography. The devised technique involves the step gradient elution with 0.2N citrate buffer (pH3-22, 4. 25) and 0.3 M borate buffer (pH 8.0, 9.9) on a column of 2, 4-divinyl benzene polymer sulfonated cation exchange resin and a high sensitive fluorescence reaction for amino acids by o-phthalaldehyde. The method described here is simple, time-saving, accurate and reproducible compared with the previous method. For the clinical purposes, plasma specimenes of 75 adult, 58 children and cerebrospinal fluid specimenes of 18 children were subjected to the analysis.
We have deveioped a new method for the quantitative determination of urinary prote in whlch was based on TCA turbidity reaction and nephelometry. This method has some merits, that is little turbidity difference between albumin and globulin, good iinearity, and using small amount of samples. From these data, it is suggested that this method will be used widely as routine urinalysls in future