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Abstract. Frings, Christopher S., and Dunn, Ralph T.: A colorimetric method
for determination of total serum lipids based on the sulfo-phospho-vanillin
THE WIDELY-ACCEPTED METHODS for mea- The phenol turbidity method of Kundel
surement of total serum lipids by extrac- and associates 5 for assay of total lipids is
tion and subsequent gravimetric analy- rapid, but was shown by Cheek and Wease 2
sis 4 ' 7 are tedious and time-consuming. A to compare unfavorably with the gravimet-
simple, rapid, reliable method for the as- ric method.
say of serum lipids which has the analytical A method based upon the oxidation of
integrity of the gravimetric method is an alcohol-ether extract of serum by a
needed. Several basic approaches to this K 2 Cr 2 0 7 -H 2 S0 4 reagent and subsequent
problem have been made. determination of the reduced chromium
One approach is to perform cholesterol, ion has been described by Bragdon. 1 This
phospholipid, and triglyceride assays, and method is time-consuming because the sol-
then calculate the total lipid concentration vent must be evaporated to dryness at 60 C.
from the values of the lipid fractions.2 This In addition, determinations of cholesterol
approach has the disadvantage of being and phosphatides are necessary so that the
very time-consuming because three inde- corresponding oxidation factors may be ap-
pendent chemical determinations are re- plied.
quired, and in addition it is necessary to Drevon and Schmit 8 reported on the
accept the often-incorrect assumption that color reaction given by lipids with vanillin
73% of total cholesterol represents choles- in a medium of sulfuric acid and phos-
terol esters. phoric acid. Postma and Stroes ° mentioned
a sulfo-phospho-vanillin method for serum
Received June 23, 1969; accepted for publication lipids, but the details were not described.
August. 10. I'OO'.l.
Presented in part at the 21st National Meeting of In this paper we describe an improved
the American Association of Clinical Chemists, Au-
gust, 1969, in Denver, Colorado. colorimetric method, based on the sulfo-
89
90 FRINGS AND DUNN Vol. 53
findings obtained from the comparison of secobarbital necessary to cause color for-
the colorimetric method, using olive oil as mation are incompatible with life.
the standard, with the gravimetric method The small volume of serum required
(Table 1). Olive oil, triolein, oleic acid, (0.1 ml.), in addition to the simplicity,
linoleic acid, linolenic acid and cholesterol speed, and reliability of the proposed
react quantitatively in the method. Either method, make it suitable for large-scale
olive oil, which in our opinion is a suit- analyses as well as for analyzing pediatric
able standard, or serum which has been as- samples.
sayed previously by a gravimetric-extraction
method can be used. References
The exact chemistry involved in this 1. Bragdon, J. H.: Colorimetric determination of
blood lipides. J. Biol. Chem., 190: 513-517,
method, just as in many methods in lipid 1951.
chemistry, is not completely understood. 2. Cheek, C. S., and Wease, D. F.: A summation
tcchnic for serum total lipids. Clin. Chem., 15:
It appears that a carbon-to-carbon double 102-107, 1969.