P. Benlian et al., Comparison of a new method for the direct and simultaneous assessment of LDL- and HDL-cholesterol with ultracentrifugation and established methods, CLIN CHEM, 46(4), 2000, pp. 493-505
Background: Automated electrophoresis combined with enzymatic cholesterol s
taining might improve routine assessment of LDL- and HDL-cholesterol (LDLC
and HDLC), as an alternative to the Friedewald equation and precipitation.
A new method (Hydrasys; SEBIA) that adapts the cholesterol esterase/cholest
erol oxidase reaction within urea-free gels was evaluated.
Methods: Fresh sera from 725 subjects (512 dyslipidemics) were analyzed by
electrophoresis, in parallel with sequential ultracentrifugation, beta-quan
tification, calculation, and precipitation.
Results: Electrophoresis was linear up to 4 g/L cholesterol, with a detecti
on limit of 0.042 g/L cholesterol/band. Within-run, between-run, between-ba
tch, and between-operator imprecision (CVs) were 1.6%, 2.0%, 1.5%, and 2.7%
for LDLC, and 3.9%, 4.3%, 5.5%, and 4.9% for HDLC, and remained unchanged
up to 6.3 g/L plasma triglycerides (TGs). Precision decreased with very low
HDLC (<0.25 g/L). Serum storage far 3-7 days at +4 or -80 OC did not inter
fere significantly with the assay. Agreement with beta-quantification was s
table for LDLC up to 5.07 g/L (r = 0.94), even at TG concentrations >4 g/L
(r = 0.91). Bias (2.88% +/- 12%) and total error (7.84%) were unchanged at
TG concentrations up to 18.5 g/L. Electrophoresis predicted National Choles
terol Education Program cut-points with <0.04 g/L error, exactly and approp
riately classified 79% and 96% of the subjects, and divided by 2.4 (all sub
jects) and 5.8 (TGs >1.5 g/L) the percentage of subjects underestimated by
calculation. One-half of the patients with TGs >4 g/L had LDLC >1.30 g/L. F
or HDLC, correlation was better with precipitation (r = 0.87) than ultracen
trifugation (r 0.76). Error (-0.10% +/- 26%) increased when HDLC decreased
(<0.35 g/L). Direct assessment of the LDLC/HDLC ratio detected 45% more hig
h-risk subjects than the calculation/precipitation combination.
Conclusions: Electrophoresis provides reliable quantification of LDLC, impr
oving precision, accuracy, and concordance over calculation, particularly w
ith increasing plasma TGs. Implementation of methods to detect low choleste
rol concentrations could extend the applications for HDLC assessment. (C) 2
000 American Association for Clinical Chemistry.