Automated determination of serum total cholesterol

1964 ◽  
Vol 10 (4) ◽  
pp. 381-384 ◽  
Author(s):  
J.B. Levine ◽  
B. Zak
Keyword(s):  
Total Cholesterol ◽  
Serum Total Cholesterol ◽  
Automated Determination

An Improved Automated Determination of Serum Total Cholesterol with a Single Color Reagent

1966 ◽  
Vol 12 (10) ◽  
pp. 681-689 ◽  
Author(s):  
Walter D Block ◽  
K John Jarrett ◽  
Jacob B Levine
Keyword(s):  
Total Cholesterol ◽  
Ferric Chloride ◽  
Flow Cell ◽  
Serum Total Cholesterol ◽  
Color Reagent ◽  
In Series ◽  
Tubular Flow ◽  
Automated Determination ◽  
Heating Bath

Abstract An anhydrous color reagent containing ferric chloride is pumped through Tygon tubing to a glass coil in a 95° heating bath. Manually prepared serum extracts (1:20 isopropanol dilution) are presented to the stream of preheated color reagent, and the two are passed through three mixing coils, connected in series. The absorbance of the resultant color is determined at 550 mµ in a 15 mm. tubular flow-cell. The improved N-automated procedure gave values of greater precision (5.6%) for total cholesterol determined in replicate samples from sera pools than the N-automated procedure which lacked precision (27.7%). The values found in 60 individual serums are within 6.0% of total cholesterol values determined by the Abell et al. method.

Modification of an Automated Procedure for Serum Cholesterol, which Permits the Quantitative Estimation of Cholesterol Esters

1971 ◽  
Vol 17 (3) ◽  
pp. 229-230 ◽  
Author(s):  
A L Siegel ◽  
B C Bowdoin
Keyword(s):  
Total Cholesterol ◽  
Reaction Temperature ◽  
Cholesterol Ester ◽  
Quantitative Estimation ◽  
Serum Total Cholesterol ◽  
Cholesterol Esters ◽  
Color Reagent ◽  
Ester Moiety ◽  
Automated Determination

Abstract Changes are described in AutoAnalyzer Methodology N-24a, for the automated determination of serum total cholesterol, that increase the reaction temperature of the mixture of sample and acid color reagent. The modified procedure eliminates the inaccuracies that attended the original methodology and allows the cholesterol ester moiety in the serum total cholesterol to be quantitated.

Method for Fully Automated Determination of Total Cholesterol in Blood Serum, Including Saponification and Extraction

1968 ◽  
Vol 14 (10) ◽  
pp. 960-966 ◽  
Author(s):  
J van der Honing ◽  
C C Saarloos ◽  
J Stip
Keyword(s):  
Carbon Tetrachloride ◽  
Blood Serum ◽  
Total Cholesterol ◽  
Standard Error ◽  
New Method ◽  
Automated Method ◽  
The Relationship ◽  
Cholesterol Determination ◽  
Automated Determination

Abstract A fully automated method has been developed for the determination of total cholesterol in blood serum, using the AutoAnalyzer system. According to the new method, based on the cholesterol determination of Abell et al. (1), the serum sample is saponified and subsequently extracted with carbon tetrachloride. After treatment with Liebermann-Burchard reagent, the amount of cholesterol is determined at 630 nm. The method can be used for free and esterified cholesterol because saponification is carried out. The relationship between the new method and that of Abell et al. is linear. The correlation coefficient is 0.98 and the standard error 1.5%. According to the new method, 30 samples can be analyzed per hour.

Reagent for the enzymatic determination of serum total cholesterol with improved lipolytic efficiency.

1983 ◽  
Vol 29 (6) ◽  
pp. 1075-1080 ◽  
Author(s):  
J Siedel ◽  
E O Hägele ◽  
J Ziegenhorn ◽  
A W Wahlefeld
Keyword(s):  
Total Cholesterol ◽  
Cholesterol Ester ◽  
High Performance ◽  
Serum Lipids ◽  
Serum Total Cholesterol ◽  
Enzymatic Determination ◽  
Layer Chromatography ◽  
Chloroform Methanol ◽  
Hydrolysis Of

Abstract We describe a sensitive method for quantifying the extent of cholesterol ester cleavage during enzymatic assay of total cholesterol in serum. Lipids are extracted from the assay mixture with chloroform/methanol (1/1 by vol), concentrated, then quantified by "high-performance" thin-layer chromatography. Although with conventional enzymatic reagents for determination of serum total cholesterol the hydrolysis of the cholesterol esters may be incomplete, a new enzymatic cholesterol reagent (Monotest Cholesterol, High Performance, Boehringer Mannheim) gives virtually complete cholesterol ester cleavage (i.e., greater than or equal to 99.5%). Use of this reagent with its improved lipolytic efficiency yields results for serum total cholesterol that are identical to those measured with a candidate reference procedure involving alkaline cholesterol ester saponification.

Direct Manual Determination of Serum Total Cholesterol with a Single Stable Reagent

1970 ◽  
Vol 16 (12) ◽  
pp. 980-984 ◽  
Author(s):  
D R Wybenga ◽  
V J Pileggi ◽  
P H Dirstine ◽  
John Di Giorgio
Keyword(s):  
Quantitative Determination ◽  
Total Cholesterol ◽  
Serum Cholesterol ◽  
Reference Method ◽  
Serum Total Cholesterol ◽  
Confidence Limits ◽  
Manual Method ◽  
Stable Reagent

Abstract We describe a simple, direct, and specific manual method for quantitative determination of total cholesterol in serum. In the method, which requires no extraction of the serum, a single stable reagent and 50 µl of sample is used. Serum cholesterol concentrations determined by this method did not differ statistically from those obtained by the reference method of Abell et al. Hemoglobin, bilirubin, and γ-globulin do not interfere unless present at markedly supranormal concentrations. Inter-run precision is about ±3% (95% confidence limits).

Kinetic enzymic method for automated determination of total cholesterol in serum.

1983 ◽  
Vol 29 (10) ◽  
pp. 1798-1802 ◽  
Author(s):  
R Deeg ◽  
J Ziegenhorn
Keyword(s):  
Total Cholesterol ◽  
Incubation Temperature ◽  
Cholesterol Oxidase ◽  
Fixed Time ◽  
Competitive Inhibitor ◽  
Cholesterol Concentration ◽  
Total Analysis ◽  
Single Standard ◽  
Automated Determination

Abstract We describe a rapid, kinetic, fixed-time method for determining serum total cholesterol by use of cholesterol esterase, cholesterol oxidase, and the indicator reaction with peroxidase, 4-aminophenazone, and phenol. On addition of the competitive inhibitor 3,4-dichlorophenol the Michaelis constant of cholesterol oxidase is apparently increased, which extends the linear relation between absorbance change and cholesterol concentration to 20.7 to 25.9 mmol/L, depending on the analyzer being used. For calibration, a single standard is used. Total analysis time is in the range of 80 to 210 s. Incubation temperature is 25 degrees C or 37 degrees C. The single-reagent procedure has been adapted to three different centrifugal analyzers and to the Eppendorf ACP 5040 analyzer. It yields precise and accurate results and is insensitive to potential interferences.

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