An Automated Method for Measuring Creatine Phosphokinase Activity in Serum

1970 ◽  
Vol 16 (4) ◽  
pp. 294-299 ◽  
Author(s):  
Ronald K Wright ◽  
Roy L Alexander
Keyword(s):  
Enzyme Activity ◽  
Enzyme Activities ◽  
Creatine Phosphokinase ◽  
Creatine Phosphate ◽  
Enzyme Concentration ◽  
Magnesium Ion ◽  
Manual Method ◽  
Automated Method ◽  
Automated Procedures

Abstract We describe a procedure for automating the determination of creatine phosphokinase (CPK) activity in serum by use of the AutoAnalyzer. Enzyme activity is determined by measuring the creatine phosphate formed from the CPK-catalyzed reaction of creatine with adenosine triphosphate (ATP). The sensitivity of the automated procedure was comparable to that of the manual method. By use of the most favorable concentrations of creatine, ATP, and magnesium ion in the substrate, a linear relationship was obtained between enzyme concentration and enzyme activities up to 600 mU, representing a sixfold improvement over that obtained by the manual method. The degree of correlation between results obtained by the manual and automated procedures is shown.

Adaptation of an Alkaline Phosphatase Method for Automatic Colorimetric Analysis

1959 ◽  
Vol 5 (2) ◽  
pp. 119-126 ◽  
Author(s):  
Walton H Marsh ◽  
Benjamin Fingerhut ◽  
Elaine Kirsch
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Standard Deviation ◽  
Enzyme Concentration ◽  
Mean Value ◽  
Colorimetric Analysis ◽  
Manual Method ◽  
Automated Method ◽  
The Mean ◽  
Standard Deviations

Abstract The alkaline phosphatase method of Kind and King was adapted to an automated recording colorimeter. The precision of the automated method (1 standard deviation as per cent of the mean value) was ±1.7 and for the manual method ±3.6 per cent. The color produced was proportional to the enzyme concentration by both methods, and recoveries of added phenol were satisfactory. In more than 150 serum specimens surveyed for enzyme activity, over 95 per cent of the results (2 standard deviations) of the 2 methods in the range 3.4-129 agree to within ±2.8 King-Armstrong units/1OO ml.

An Automated Method for the Determination of Estrogens in Pregnancy

1968 ◽  
Vol 14 (10) ◽  
pp. 1010-1022 ◽  
Author(s):  
D Ua Conaill ◽  
G G Muir
Keyword(s):  
Turnover Rate ◽  
Good Recovery ◽  
Manual Method ◽  
Color Development ◽  
Automated Method ◽  
Combined Operation ◽  
In Pregnancy

Abstract Ittrich’s method for the determination of total estrogens in urine (1) was adapted for automation on the AutoAnalyzer, using a continuous digester module for the combined operation of phase exchange and color development. The automated method gave good recovery and reproducibility, and correlated well with the manual method. It increased the capacity of this laboratory for estrogen determinations, improved the turnover rate, and reduced the labor involved to a minimum.

Responses of the phosphatase activity of the lichen Cladina rangiferina to various environmental factors including metals

1979 ◽  
Vol 57 (14) ◽  
pp. 1534-1540 ◽  
Author(s):  
I. Lane ◽  
K. J. Puckett
Keyword(s):  
Enzyme Activity ◽  
Environmental Factors ◽  
Phosphatase Activity ◽  
Enzyme Activities ◽  
Enzyme Concentration ◽  
Acidic Ph ◽  
Nitrophenyl Phosphate ◽  
Cladina Rangiferina ◽  
Reduced Activity ◽  
Menten Constant

The characteristics of phosphatase activity of Cladina rangiferina (L.) Harm, have been studied. Calculations of enzyme activities were based on the liberation of p-nitrophenol from p-nitrophenyl phosphate. The phosphatase activity was found to be linear both with increasing sample size (enzyme concentration) and increasing time, showed highest activity at acidic pH, and had a Michaelis–Menten constant of 8.9 × 10−3 M. The enzyme activity was maximal in the range 61 ± 10 °C, was independent of light, and was completely eliminated by boiling the thalli. Various cations and anions were tested for their effect; uranyl and vanadyl ions inhibited activity by 60% whereas copper, nickel, and silver enhanced activity by 10%. The anions biselenite, cyanide, fluoride, molybdate, phosphate, and vanadate all greatly reduced activity (≥ 50%). Phosphatase activity was demonstrated in other lichen species.

A Simple Automated Colorimetric Method for Determination of N-Acetyl- β-d-Glucosaminidase

1982 ◽  
Vol 19 (3) ◽  
pp. 157-159 ◽  
Author(s):  
Joan C Moore ◽  
Jacqueline E Morris
Keyword(s):  
Enzyme Activity ◽  
Calcium Chloride ◽  
Coefficient Of Variation ◽  
Colorimetric Method ◽  
Plasma Samples ◽  
Automated Method

An automated method for the determination of N-acetyl-β-d-glucosaminidase in serum or plasma using p-nitrophenol ( pNP) N-acetyl-β-d-glucosamine as substrate and a Pye Unicam AURA system is described. Normal samples had activities of 853 ± 146 (SD) nmol pNP liberated/ml/h, with intra-assay coefficient of variation 1·2% and inter-assay coefficient of variation 1·6%. Inhibition of enzyme activity by heparin in plasma samples can be reversed by the addition of calcium chloride to the buffer.

Automated Method for the Determination of Serum Creatine Kinase Activity

1967 ◽  
Vol 13 (3) ◽  
pp. 233-241 ◽  
Author(s):  
Gerard A Fleisher
Keyword(s):  
Creatine Kinase ◽  
Alkaline Solution ◽  
Kinase Activity ◽  
Adenosine Diphosphate ◽  
Creatine Phosphate ◽  
Creatine Kinase Activity ◽  
Serum Creatine Kinase ◽  
Colorimetric Determination ◽  
Automated Method

Abstract An automated (AutoAnalyzer) method for the colorimetric determination of creatine kinase activity in serum is described. This method includes reactivation of creatine kinase with cysteine, incubation of the active enzyme with creatine phosphate and adenosine diphosphate at 37.5°, and subsequent inactivation of enzyme and binding of cysteine by phenylmercuric borate. The enzymatically produced creatine is dialyzed against a solution of diacetyl and reacted with α-naphthol in an alkaline solution. The absorbance of the colored end product is measured at 550 mµ. Individual blanks are determined in the absence of adenosine diphosphate. Comparison of results obtained by this method and a manual procedure shows satisfactory agreement.

Automated Simultaneous Determination of p-Aminohippurate and Creatinine in Plasma or Urine

1974 ◽  
Vol 20 (3) ◽  
pp. 348-352 ◽  
Author(s):  
C K Parekh ◽  
J Kirpan ◽  
A Peterson ◽  
G L Hassert ◽  
B F Murphy
Keyword(s):  
Simultaneous Determination ◽  
Urine Samples ◽  
Manual Method ◽  
Automated Method ◽  
Discrete Sample ◽  
Beer's Law ◽  
Plasma And Urine Samples ◽  
Beer’S Law

Abstract Methods have been developed for using the Beckman "Discrete Sample Analyzer" (DSA-560), for automated simultaneous determinations of p-aminohippurate (PAH) and creatinine in the same 50- or 10-µl samples of plasma or urine, respectively, at the rate of 80 samples per hour. In determinations of PAH, a single reagent, p-dimethylaminocinnamaldehyde in 0.1 mol/liter HCl, was added to a protein-free filtrate from plasma or urine. The intensity of the color, measured at 550 nm, obeyed Beer’s law for PAH concentrations from 0.005 to 5.0 g/liter. Data obtained for the same plasma and urine samples by the manual method, were, in general, within ±5% of the results obtained by the automated method. Creatinine was simultaneously determined by adapting the method of Taussky to the DSA-560 instrument; the intensity of the color, measured at 510 nm, obeyed Beer's law for creatinine concentrations ranging from 0.005 to 5.0 g/liter. Data obtained for the same plasma and urine samples by the manual method, were, in general, within ± 5% of the results obtained by the automated method.

scholarly journals A RAPID METHOD FOR THE DETERMINATION OF PROTEOLYTIC ACTIVITIES OF ENZYME PREPARATIONS

1948 ◽  
Vol 32 (1) ◽  
pp. 17-24 ◽  
Author(s):  
Bacon F. Chow ◽  
Mary-Ann Peticolas
Keyword(s):  
Enzyme Activity ◽  
Trichloroacetic Acid ◽  
Enzyme Concentration ◽  
Cent Solution ◽  
Enzyme Preparations ◽  
Proteolytic Activities ◽  
Activity Of Enzymes ◽  
Different Sources ◽  
Rate Of Activation

A method has been described for the determination of proteolytic activities of enzyme preparations using casein as substrate. The rate of digestion is proportional to the enzyme concentration used. This relationship is utilized as a measure of the enzyme activity. One unit of activity is defined as the amount which is required to digest casein in 15 minutes at 37.5°C. so that 50 per cent of the protein in 1 ml. of 0.25 per cent solution is not precipitable by trichloroacetic acid. This method has been used to determine the activity of enzymes from different sources and also used to follow the rate of activation of enzymes.

scholarly journals An Automated Method for the Determination of Intestinal Disaccharidase and Glucoamylase Activities

2006 ◽  
Vol 2006 ◽  
pp. 1-4 ◽  
Author(s):  
Zhaoping He ◽  
Laura Bolling ◽  
Dalal Tonb ◽  
Tracey Nadal ◽  
Devendra I. Mehta
Keyword(s):  
Quality Control ◽  
Automated System ◽  
Internal Quality ◽  
Perfect Agreement ◽  
Manual Method ◽  
Automated Method ◽  
Reaction Volumes ◽  
High Consistency ◽  
Larger Sample

Determination of disaccharidase and glucoamylase activities is important for the diagnosis of intestinal diseases. We adapted a widely accepted manual method to an automated system that uses the same reagents reaction volumes, incubation times, and biopsy size as the manual method. A dye was added to the homogenates as the internal quality control to monitor the pipetting precision of the automated system. When the automated system was tested using human intestinal homogenates, the activities of all the routinely tested disaccharidases, including lactase, maltase, sucrase, and palatinase, as well as the activity of glucoamylase, showed perfect agreement with the manual method and were highly reproducible. The automated analyzer can perform the same routine assays of disaccharidases and glucoamylase with high consistency and accuracy and reduce testing costs by performing a larger sample size with the same number of staff. Additional developments, such as barcoding and built-in plate reading, would result in a completely automated system.

Automated Determination of Serum Ceruloplasmin Activity with o-Dianisidine Dihydrochloride as Substrate

1975 ◽  
Vol 21 (6) ◽  
pp. 757-759 ◽  
Author(s):  
Karl H Schosinsky ◽  
Peter Lehmann ◽  
Myrton F Beeler
Keyword(s):  
Coefficient Of Variation ◽  
Serum Ceruloplasmin ◽  
Manual Method ◽  
Automated Method ◽  
Enzymatic Determination ◽  
Ceruloplasmin Activity ◽  
Automated Determination

Abstract An automated method for the enzymatic determination of ceruloplasmin with o-dianisidine dihydrochioride as substrate is described. The method enables the measurement of 30 samples per hour with a coefficient of variation (day-to-day) of 2.8%. Results correlate well (r = 0.99) with those obtained by the corresponding manual method.

Determination of Serum or Plasma Glucose on the "AutoAnalyzer II" by Use of the Hexokinase Reaction

1972 ◽  
Vol 18 (3) ◽  
pp. 299-300 ◽  
Author(s):  
G M Widdowson ◽  
J R Penton
Keyword(s):  
Plasma Glucose ◽  
Good Precision ◽  
Manual Method ◽  
Automated Method

Abstract An automated method is described for determination of glucose in serum or plasma by use of the hexokinase reaction. The method has good precision and the results correlate well with those from a manual method in which the same reaction is used.

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