Dextran sulfate-Mg2+ precipitation procedure for quantitation of high-density-lipoprotein cholesterol.

1982 ◽  
Vol 28 (6) ◽  
pp. 1379-1388 ◽  
Author(s):  
G R Warnick ◽  
J Benderson ◽  
J J Albers
Keyword(s):  
High Density Lipoprotein ◽  
High Density Lipoprotein Cholesterol ◽  
Dextran Sulfate ◽  
Density Lipoprotein ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
Precipitation Procedure

scholarly journals Three routine methods for measuring high-density lipoprotein cholesterol compared with the Reference Method

1996 ◽  
Vol 42 (5) ◽  
pp. 738-743 ◽  
Author(s):  
N Harris ◽  
V Galpchian ◽  
N Rifai
Keyword(s):  
High Density Lipoprotein ◽  
High Density Lipoprotein Cholesterol ◽  
Dextran Sulfate ◽  
Direct Method ◽  
Density Lipoprotein ◽  
Reference Method ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
Direct Assay ◽  
Wide Range

Abstract We compared the performance of three methods for quantifying high-density lipoprotein cholesterol (HDL-C) with the Reference Method for HDL-C, using samples with a wide range of triglyceride (TG) concentrations (290-18000 mg/L). All three comparison assays-- utilizing a magnetic dextran sulfate precipitating reagent, a direct method, and a standard MgCl2-dextran sulfate reagent--were precise, with a run-to-run CV of less than or equal to 4.1%. However, the systematic error of these assays exceeded the National Cholesterol Education Program (NCEP) performance goal of less than or equal to 10% in half of the concentration ranges tested. Nevertheless, the total error of the assays generally meets the current 22% limit set by the NCEP. Although both the magnetic dextran sulfate precipitation reagent and the direct assay can be performed more rapidly than the MgCl2-dextran sulfate assay, the direct assay involves no sample preparation and requires only 4 microL of sample excluding the dead space. Although precipitation is frequently inadequate with the MgCl2-dextran sulfate reagent at TG concentrations >6000 mg/L, both the magnetic and the direct reagent show no interference from high TG concentrations as great as 18 000 mg/L.

Comparison of the Du Pont aca and Dow methods for determination of high-density lipoprotein cholesterol.

1984 ◽  
Vol 30 (1) ◽  
pp. 127-129 ◽  
Author(s):  
N N Rehak ◽  
R J Elin ◽  
R Chesler ◽  
E Johnson
Keyword(s):  
High Density Lipoprotein ◽  
Disease Control ◽  
Total Cholesterol ◽  
High Density Lipoprotein Cholesterol ◽  
Dextran Sulfate ◽  
Density Lipoprotein ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
Serum Samples

Abstract We compared the Du Pont aca (phosphotungstate-enzymic cholesterol) and the Dow (dextran sulfate/Mg2+-enzymic cholesterol) methods for the determination of high-density lipoprotein cholesterol (HDLC) and total cholesterol in serum from 113 patients. The aca results for both total cholesterol and HDLC were significantly greater (p less than 0.0001) than the Dow results, the aca method overestimating the HDLC concentration (mean recovery 107.2% in serum samples with values assigned by the Centers for Disease Control). The precision of the aca method for HDLC was essentially the same as that of the Dow method. Bilirubin (up to 0.17 g/L), hemoglobin (up to 4 g/L), and slight lipemia (triglycerides up to 5.4 g/L) did not interfere with the aca method.

Comparison of improved precipitation methods for quantification of high-density lipoprotein cholesterol.

1985 ◽  
Vol 31 (2) ◽  
pp. 217-222 ◽  
Author(s):  
G R Warnick ◽  
T Nguyen ◽  
A A Albers
Keyword(s):  
Polyethylene Glycol ◽  
High Density Lipoprotein ◽  
High Density Lipoprotein Cholesterol ◽  
Dextran Sulfate ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
High Density ◽  
Protein Precipitation ◽  
Lipoprotein Cholesterol ◽  
Lipid Research

Abstract We compared the standard Lipid Research Clinics heparin-Mn2+ (46 mmol/L) method and five improved precipitation methods for quantification of high-density lipoprotein (HDL) cholesterol. Three of these methods--a dextran sulfate-Mg2+ procedure, reported as a Selected Method, a modified heparin-Mn2+ (92 mmol/L) method, and a modified phosphotungstate-Mg2+ procedure--all gave similar results. Three other methods--the standard heparin-Mn2+ (46 mmol/L) method and two polyethylene glycol methods (75 g/L or pH 10 reagent at 100 g/L final concentrations)--gave slightly higher values for HDL cholesterol. Addition of NaCl or glucose to specimens did not significantly change protein precipitation. In terms of sedimentation effectiveness with hypertriglyceridemic specimens, the methods were ranked in the following order: polyethylene glycol (pH 10, 100 g/L) greater than dextran sulfate-Mg2+ greater than heparin-Mn2+ (92 mmol/L) = polyethylene glycol (75 g/L) greater than phosphotungstate-Mg2+ greater than heparin-Mn2+ (46 mmol/L).

Comparison of current methods for high-density lipoprotein cholesterol quantitation.

1979 ◽  
Vol 25 (4) ◽  
pp. 596-604 ◽  
Author(s):  
G R Warnick ◽  
M C Cheung ◽  
J J Albers
Keyword(s):  
Polyethylene Glycol ◽  
High Density Lipoprotein ◽  
High Density Lipoprotein Cholesterol ◽  
Dextran Sulfate ◽  
Density Lipoprotein ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
Precipitation Method ◽  
Apoprotein B ◽  
Polyethylene Glycol 6000

Abstract We compared six precipitation methods for high-density-lipoprotein cholesterol quantitation with an ultracentrifugation method, the accuracy of which was improved by correcting for 4% manipulative loss in the d greater than 1.063 fractions. For purposes of comparison, the apoprotein B-associated cholesterol (average 56 mg/L) measured by immunoassay in the d greater than 1.063 fractions was subtracted. In 65 plasma samples from men, women, and children, a heparin-Mn2+ procedure with Mn2+ at 46 mmol/L produced results slightly higher (+16 mg/L), while results with Mn2+ at 92 mmol/L averaged slightly lower (-8 mg/L) than the comparison ultracentrifuge method. Results that were about 5% low were obtained by the dextran sulfate 500-Mg2+ (-25 mg/L) and phosphotungstate-Mg2+ (-31 mg/L) methods. A heparin-Ca2+ method produced results 10% high (+58 mg/L). Results by a polyethylene glycol-6000 precipitation method were 12% low (-64 mg/L). Precision was better with the two heparin-Mn2+ and the dextran sulfate 500-Mg2+ procedures, with CVs of 4%, intermediate with phosphotungstate-Mg2+ and polyethylene glycol-6000 (CV 6-7%), and poorest with heparin-Ca2+ (CV 10%). Precipitation by phosphotungate-Mg2+ appeared more sensitive to reagent concentration and temperature variations than either the heparin-Mn2+ or dextran sulfate 500-Mg2+ methods. We conclude that these precipitation methods are not equivalent, but give rise to significant systematic differences in high-density-lipoprotein cholesterol quantitation.

Cholesterol in high-density lipoprotein: use of Mg2+/dextran sulfate in its enzymic measurement.

1978 ◽  
Vol 24 (6) ◽  
pp. 931-933 ◽  
Author(s):  
P R Finley ◽  
R B Schifman ◽  
R J Williams ◽  
D A Lichti
Keyword(s):  
High Density Lipoprotein ◽  
High Density Lipoprotein Cholesterol ◽  
Dextran Sulfate ◽  
Density Lipoprotein ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
Cholesterol Concentration ◽  
High Density Lipoproteins ◽  
Low Density ◽  
Very Low Density Lipoproteins

Abstract We describe a method for measuring high-density lipoprotein cholesterol. MgCl2 and dextran sulfate are used to precipitate all low-density and very-low-density lipoproteins. The supernate contains only high-density lipoproteins, the cholesterol concentration of which is estimated by an enzymic method, with a discrete analyzer (Abbott Bichromatic Analyzer). Concentration and instrument response are linearly related to 50 mg/liter. The precision of the method is excellent in the range of clinical interest (100 to 1000 mg of cholesterol per liter). The precision and efficiency of the precipitation are shown at various concentrations of high-density lipoprotein cholesterol. The method was compared to that of two laboratories in the Cooperative Lipoprotein Phenotyping Study group by testing a number of split samples, and agreement was good.

scholarly journals Precipitation methods for high-density lipoprotein cholesterol measurement compared, and final evaluation under routine operating conditions of a method with a low sample-to-reagent ratio

1997 ◽  
Vol 43 (4) ◽  
pp. 663-668 ◽  
Author(s):  
Pierre N M Demacker ◽  
Marja Hessels ◽  
Helga Toenhake-Dijkstra ◽  
Henk Baadenhuijsen
Keyword(s):  
High Density Lipoprotein ◽  
High Density Lipoprotein Cholesterol ◽  
Dextran Sulfate ◽  
Total Error ◽  
Density Lipoprotein ◽  
Volume Ratio ◽  
High Density ◽  
Operating Conditions ◽  
Lipoprotein Cholesterol ◽  
Peg 6000

Abstract We evaluated six precipitation methods for high-density lipoprotein cholesterol (HDL-chol) determination: the heparin/Mn2+precipitation reagent method (Hep), two variants of the phosphotungstic acid/Mg2+ method (Tung-L and Tung-B), the dextran sulfate 50 000/Mg2+ method (Dex), the PEG 6000 method (PEG), and the PEG 6000/dextran sulfate 15 000 (PEG/Dex) method. The Tung-B and PEG/Dex precipitation methods have a low sample/precipitation reagent volume ratio (<0.4). The Tung-B, Dex, PEG, and PEG/Dex methods gave similar values, averaging within 0.1 mmol/L of each other, showing that the precipitation selectivity of these methods is comparable. The precipitation efficiency of Tung-B and Peg/Dex, however, was superior. Ultrafiltration of the supernatants was needed only at triglyceride concentrations >16.4 mmol/L (undiluted sample) or >28.0 mmol/L (sample diluted twofold); however, ultrafiltration without dilution was the most accurate method. Results of Tung-B under routine conditions (33 technicians) agreed well with those of the PEG method for 406 normo- and hyperlipidemic plasma samples. By comparison with the HDL-chol method from the Centers for Disease Control and Prevention, the Tung-B method showed a total error of 10.6%, which fulfills the criteria of the National Cholesterol Education Program for HDL-chol analysis. In conclusion, with motivated personnel, Tung-B is a reliable, cost-effective method for routine HDL-chol analysis.

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