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.

Improved method for determination of high-density-lipoprotein cholesterol I. Isolation of high-density lipoproteins by use of polyethylene glycol 6000.

1981 ◽  
Vol 27 (3) ◽  
pp. 371-374 ◽  
Author(s):  
C Izzo ◽  
F Grillo ◽  
E Murador
Keyword(s):  
Polyethylene Glycol ◽  
High Density Lipoprotein ◽  
High Density Lipoprotein Cholesterol ◽  
Density Lipoprotein ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
High Density Lipoproteins ◽  
Good Correspondence ◽  
Simple Method ◽  
Polyethylene Glycol 6000

Abstract We describe a modified method of precipitating low- and very-low-density lipoproteins with polyethylene glycol, for quantitation of high-density lipoprotein cholesterol. A 100 g/L concentration of polyethylene glycol 6000 in a buffered (pH 10) solution is used. Under these conditions precipitation of beta-lipoproteins is complete. Values for HDL cholesterol as measured with this method fully correspond to those obtained on separating the lipoproteins by ultracentrifugation or with heparin-Mn2+, 46 mmol/L; are slightly higher than those obtained with heparin-Mn2+, 92 mmol/L; and are significantly higher than those determined after precipitation with polyethylene glycol, 100 g/L at pH 8.0. Comparison with results by the ultracentrifugation method showed excellent correlation (r = 0.957) with a good correspondence of results (slope 1.065 and y-intercept -22.231); comparison with the heparin-Mn2+, 46 mmol/L, method gave r = 0.998 with slope of 0.985 and y-intercept 2.963. The method is unaffected by precipitation and centrifugation time and temperature (up to 24 h with 4-22 degrees C temperature for precipitation and between 10 and 30 min with 4-22 degrees C temperature for centrifugation). This extremely simple method is particularly suitable for routine analyses of many samples, even in small laboratories.

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).

A study of the use of polyethylene glycol in estimating cholesterol in high-density lipoprotein.

1980 ◽  
Vol 26 (13) ◽  
pp. 1775-1779 ◽  
Author(s):  
P N Demacker ◽  
A G Hijmans ◽  
H E Vos-Janssen ◽  
A van't Laar ◽  
A P Jansen
Keyword(s):  
Polyethylene Glycol ◽  
High Density Lipoprotein ◽  
Reference Values ◽  
High Density Lipoprotein Cholesterol ◽  
Density Lipoprotein ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
High Density Lipoproteins ◽  
Polyethylene Glycol 6000

Abstract We studied polyethylene glycol 6000 precipitation of lipoproteins other than high-density lipoproteins, before cholesterol is estimated in the supernate. Other lipoproteins in the supernatant fractions were detected by using rocket immunoelectrophoresis. A polyethylene glycol concentration of 75 g/L in the final mixture appeared to be optimal, and results agreed with those obtained by ultracentrifugation. Differences in serum pH, use of polyethylene glycol from different suppliers, or the presence of ethylenediaminetetraacetate resulted in values that differed significantly (by 40 to 60 mumol/L) from the reference values. Polyethylene glycol did not interfere in four different methods for determination of cholesterol. In combination with an enzymic cholesterol method, the polyethylene glycol method appeared to be very precise, even when lipemic sera (triglycerides up to 5.5 mmol/L) were analyzed that had diminished high-density lipoprotein cholesterol values. We consider this method a method of choice, especially when lipemic sera are tested and enzymic cholesterol analysis is used.

An enzymic and centrifugal method for estimating high-density lipoprotein cholesterol.

1979 ◽  
Vol 25 (2) ◽  
pp. 325-327 ◽  
Author(s):  
J K Allen ◽  
W J Hensley ◽  
A V Nicholls ◽  
J B Whitfield
Keyword(s):  
Polyethylene Glycol ◽  
High Density Lipoprotein ◽  
High Density Lipoprotein Cholesterol ◽  
Density Lipoprotein ◽  
Final Concentration ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
Precipitation Method ◽  
Low Density ◽  
Normal Ranges

Abstract Enzymic measurement of high-density lipoprotein cholesterol with a centrifugal analyzer is described. We used polyethylene glycol (Mr 6000), final concentration 100 g/L, to precipitate low-density and very-low-density lipoproteins, thereby eliminating the difficulties of the commonly used heparin/Mn2+ precipitation method and facilitating the use of ethylenediaminetetraacetate-stabilized plasma. As measured by rocket immunoelectrophoresis, this final concentration of polyethylene glycol completely precipitates beta-lipoproteins, leaving the alpha-lipoproteins in solution. Between-run reproducibility (CV) was 3.6%, within-run reproducibility (CV) 0.8%. Reagent costs currently are $US 0.13 per test and large numbers of samples can be handled conveniently. Normal ranges were compiled for 539 men and 444 women. The high-density lipoprotein cholesterol for men was 1.20 +/- 0.31 (SD) mmol/L and for women 1.52 +/- 0.38 (SD) mmol/L.

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.

scholarly journals Evaluation of two homogeneous methods for measuring high-density lipoprotein cholesterol

1997 ◽  
Vol 43 (6) ◽  
pp. 1048-1055 ◽  
Author(s):  
Yi-Chang Huang ◽  
Jau-Tsuen Kao ◽  
Keh-Sung Tsai
Keyword(s):  
Polyethylene Glycol ◽  
High Density Lipoprotein ◽  
High Density Lipoprotein Cholesterol ◽  
Phosphotungstic Acid ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
Homogeneous Assays ◽  
Better Than

Abstract We evaluated the performance of two homogeneous assays for quantifying HDL cholesterol (HDL-C) and compared them with the phosphotungstic acid (PTA)/MgCl2 assay. Both homogeneous HDL-C assays were precise, having a within-run CV of <1.20% and a between-run CV of <4.07%. The HDL-C values (y) measured by the two homogeneous methods correlated well with those by the PTA/MgCl2 method (x): y = 1.00x + 64.98 mg/L, r = 0.987, Sy|x = 27.99 mg/L (n = 152) for the polyethylene glycol-modified enzymes/α-cyclodextrin sulfate (PEGME) assay (Kyowa), and y = 0.84x + 106.51 mg/L, r = 0.984, Sy|x = 26.10 mg/L (n = 152) for the polyanion–polymer/detergent (PPD) assay (Daiichi). The specificity of the PEGME method seemed better than that of the PPD method, as the PPD method was markedly interfered with by supplemental LDL-C. Addition of 20 g/L triglycerides produced a negative error of ∼18% in both homogeneous assays. Bilirubin and hemoglobin had little influence on the PEGME method; hemoglobin had little effect on the PPD method. Bilirubin, however, markedly decreased the readings by the PPD method. We found the PEGME assay superior to the PPD assay for routine HDL-C testing, because the PPD assay is relatively inaccurate and not specific.

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.

Sign in / Sign up

Export Citation Format

Share Document