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Joseph D. Artiss Michael W. McGowan Donald R. Strandbergh Bennie Zak 《Clinica chimica acta; international journal of clinical chemistry》1982,124(2):141-148
A choline oxidase-peroxidase coupled enzyme procedure is proposed for the determination of cholinesterase activity in human serum. This system is not only kinetic and colorimetric but is also relatively quick and simple to perform. The initial comparisons suggest that this method correlates well with a commonly used propionylthiocholine-dinitrobis-(nitro-benzoic acid) technique. Large amounts of bilirubin in the sample appear to have only minor deleterious effects on the assay. Since there are only two reagents that may be premixed, the procedure appears to be amenable to automation.The use of a mixture of sodium 2-hydroxy-3,5-dichlorobenzenesulfonate and 4-aminoantipyrene in the peroxidase catalyzed indicator reaction provides for a marked increase in sensitivity over previously reported 4-aminoantipyrene-phenol systems. This augmented sensitivity provides for a relatively large reagent to sample ratio. In addition, the reagents lend themselves toward lyophilization or ‘dry-fill’. 相似文献
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C H Konings 《Clinica chimica acta; international journal of clinical chemistry》1988,176(2):185-193
A method for estimation of arginine in 50 microliters serum was developed using commercially available arginine kinase (EC 2.7.3.3). The assay is based on the transformation of arginine and ATP into phospho-arginine and ADP by the enzyme. ADP is measured by two coupling reactions involving pyruvate kinase (EC 2.7.1.40) and lactate dehydrogenase (EC 1.1.1.27) with measurement of NADH consumption at 340 nm. The method involves preincubation of serum in the reaction medium without arginine kinase to eliminate side reactions and a kinetic rate protocol with measurements of absorbance at 60 s and 180 s. Reaction temperature is 30 degrees C. The reaction is linear up to at least 3 mmol/l of arginine. Within-batch CV is less than 3% for arginine levels above 0.75 mmol/l and the between-batch CV is 6.5% or less. The method correlates well with an automatic amino acid analyzer procedure (r = 0.983). The reference range derived from sera of 40 blood donors has been determined to be 0.06-0.20 mmol/l. 相似文献
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We evaluated an enzymatic procedure for the automated determination of serum and urinary creatinine. The procedure was sensitive and precise for serum creatinine concentrations as low as 3 mg/L, and the standard curve was linear to 200 mg/L. Measurements of creatinine in clinical serum and urine specimens agreed with those obtained by two other enzymatic methods and four picric acid methods. There was no interference from picrate-reactive substances tested. Among other substances, bilirubin (50-100 mg/L) interfered negatively with the assay, decreasing apparent creatinine concentrations by as much as 6 mg/L. The advantages of specificity, speed, and convenience make this enzymatic procedure an attractive primary or secondary method for creatinine determinations in clinical laboratories. 相似文献
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We have explored a kinetic colorimetric method for measuring gamma-glutamyltransferase (EC 2.3.2.2) activity in serum, using L-gamma-glutamyl-3,5-dibromo-4-hydroxyanilide and glycylglycine as donor and acceptor substrates. The released product, 3,5-dibromo-4-hydroxyaniline, reacts with 2,5-dimethylphenol to produce a blue quinone monoimine in the presence of ascorbate oxidase (EC 1.10.3.3). This dye has peak absorption at 610 nm, whereas the donor substrate shows negligible absorption throughout the visible spectrum. The reaction can be run with all the reagents in a single working solution with serum as starter, or with the substrate solution as starting reagent. The sample/reagent volume ratio is 1:24. Adaptation of the method to several automated instruments gave good precision in all cases. Comparison with a method in which L-gamma-glutamyl-3-carboxy-4-nitroanilide is the donor substrate showed good correlation of results (r greater than or equal to 0.987). The dynamic range of the method exceeds the upper limits of the reference intervals for men (9-33 U/L) and women (8-25 U/L) by at least 18-fold. 相似文献
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M Usategui-Gomez P Tarbutton F Yeager 《Clinica chimica acta; international journal of clinical chemistry》1973,47(3):409-415
A simple, rapid colorimetric assay for oxytocinase in pregnancy serum is described. The assay, which utilizes as the substrate, is based on the formation of an azo-dye. The assay required a 10-min incubation at 37°, followed by 6 min for color development. 相似文献
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J.F. Burd M. Usategui-Gomez 《Clinica chimica acta; international journal of clinical chemistry》1973,46(3):223-227
A simple, rapid, colorimetric assay for serum lactate dehydrogenase is described. The method requires 0.05 ml serum and a 12-min incubation at 37° and utilizes a tetrazolium salt in conjunction with a stable intermediate electron carrier (Meldola Blue). Good correlation between the colorimetric assay and a reference spectrophotometric assay was observed. 相似文献
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A colorimetric procedure for the determination of blood ammonia 总被引:9,自引:0,他引:9
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A rapid colorimetric assay for serum lactic dehydrogenase 总被引:2,自引:0,他引:2
A L Babson G E Phillips 《Clinica chimica acta; international journal of clinical chemistry》1965,12(2):210-215
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An improved colorimetric procedure for urine oxalate 总被引:6,自引:0,他引:6
A Hodgkinson A Williams 《Clinica chimica acta; international journal of clinical chemistry》1972,36(1):127-132
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A Kyaw 《Clinica chimica acta; international journal of clinical chemistry》1976,69(2):351-354
A new color reagent, 4-aminophenazone, has been introduced for iron determination. The reagent has been found to be specific and sensitive to iron and can be used for serum iron determination with good reproducibility. 相似文献
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连续监测法测定血清丙氨酸氨基肽酶 总被引:4,自引:0,他引:4
目的 建立血清丙氨酸氨基肽酶 (AAP)连续监测法。方法 以丙氨酰对硝基苯胺 (Ala pNA)为底物 ,研究AAP作用于此底物的动力学特征 ,选择最适反应条件 ,建立血清AAP测定速率法及参考值 ,同时观测肝胆疾病患者血清AAP作用于此底物的动力学特征 ,选择最适反应条件 ,建立血清AAP测定速率法及参考值 ,同时观测肝胆疾病患者血清AAP水平。结果 酶反应最适pH8 0 ,Tris HCl缓冲液的最佳浓度 15 0mmol/L ,Ala pNa浓度以 3mmol/L为宜 ,米氏常数 (Km) 0 74mmol/L ,在 8min观测时间内吸光度变化与时间成比例 ,线性范围可达 180 0U/L ,平均批内、批间CV分别为 1 0 4%和 2 0 2 %。 2 0mmol/L甘油三酯、2g/L血红蛋白、340 μmol/L胆红素对酶活力测定无干扰 ,正常成年人群参考范围 ( x± 2s) 4 1~ 77U/L。肝胆疾病组血清AAP明显高于健康对照组。结论 血清AAP活力是鉴别诊断肝胆疾病的一项特异、灵敏的指标 ;连续监测法适用于各种类型的自动化分析仪 ,便于临床推广 相似文献
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A peroxidase-coupled method for the colorimetric determination of serum triglycerides 总被引:16,自引:0,他引:16
We describe an enzymatic method for rapid, precise measurement of serum triglycerides with use of sample:reagent ratios as large as 1:200. Hydrolysis of triglycerides is catalyzed by lipase to produce glycerol and free fatty acids. The glycerol generated is then phosphorylated by adenosine 5'-triphosphate in the presence of glycerol kinase. Oxidation of the resulting glycerol 3-phosphate to produce hydrogen peroxide is catalyzed by L-alpha-glycerophosphate oxidase. An intense red chromogen is produced by the peroxidase-catalyzed coupling of 4-aminoantipyrene and sodium 2-hydroxy-3,5-dichlorobenzenesulfonate with hydrogen peroxide. This sensitive chromogen system not only permits use of unusually small sample volumes, it also facilitates a linear response to serum triglyceride concentrations up to at least 10 g/L while displaying good Ringbom (measure of accuracy) characteristics. 相似文献
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This rapid, sensitive equilibrium turbidimetric immunoassay for quantification of alpha 1-antitrypsin involves a monospecific antibody, polyethylene glycol 6000 to accelerate and enhance the immunoprecipitation reaction, and Tween 20 surfactant to decrease and stabilize the sample-blank values. Turbidity at 334 nm is measured by an automated discrete analyzer. Grossly lipemic, icteric, or hemolyzed samples can be assayed. Correlation with results by radial immunodiffusion (RID) was excellent (r = 0.97, n = 84). Analytical recovery averaged 97.7 (SD 2.9)%. Within-run CVs ranged from 1.6 to 1.9%, between-day CVs from 2.0 to 3.5%. Reference values for healthy adults (n = 147) were determined by parametric estimation (for an assumed normal distribution of untransformed data). The lower limit (g/L) with its 0.90 confidence interval is 1.23 (range 1.18-1.28), the upper limit is 2.15 (2.10-2.20), and the mean is 1.69 g/L. 相似文献
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This is a method for measuring tripeptide aminopeptidase (EC 3.4.11.4) activity in serum. L- Leucylglycylglycine is used as substrate, and the reaction is followed by monitoring the absorbance increase at 340 nm when NAD+ is reduced to NADH in the presence of an excess of leucine dehydrogenase. This principle allows kinetic determination of the enzyme without interference by carboxypeptidases. Amastatin is added to the reaction mixture to prevent nonspecific hydrolysis of the substrate catalyzed by other aminopeptidases. As final reaction concentrations we recommend (per liter): 100 mmol of Tris buffer (pH 8.2), 4.0 mmol of L- leucylglycylglycine , 10 kU of leucine dehydrogenase, 3.8 mmol of NAD+, and 85 mumol of amastatin . The assay is suited to modern enzyme analyzers and has high precision. 相似文献