Isotope dilution/mass spectrometry of serum cholesterol with (3,4-/sup 13/C)cholesterol: proposed definitive method

Journal Article · Sat Aug 01 00:00:00 EDT 1987 · Clin. Chem. (Winston-Salem, N.C.); (United States)

OSTI ID:6375377

Pelletier, O; Wright, L A; Breckenridge, W C

We describe a new gas-chromatographic/mass-spectrometric (GC/MS) isotope-dilution method for determination of serum cholesterol. The method has been fully optimized and documented to provide the high accuracy and precision expected for a Definitive Method. In the presence of (3,4-/sup 13/C)cholesterol, cholesteryl esters in serum are hydrolyzed under optimum conditions and the entire cholesterol pool is extracted and derivatized to silyl ethers. The cholesterol derivatives are resolved from other sterols by gas-liquid chromatography on a fused silica column, and selected ions characteristic of cholesterol and the (3,4-/sup 13/C)cholesterol are monitored with a GC/MS quandrupole system. We estimated the cholesterol content of samples by bracketing each sample with standards of comparable cholesterol concentration that also contained the (3,4-/sup 13/C)cholesterol. The procedure was highly reproducible (CV less than 0.5%), better accuracy and precision being obtained with (3,4-/sup 13/C)cholesterol than with heptadeuterated cholesterol. Mean values per gram of dry serum for one serum pool assayed by this method and that of the National Bureau of Standards differed by 0.5%. We conclude that the method satisfies the criteria for a Definitive Method.

• Research Organization: Health Protection Branch, Health and Welfare, Ottawa, Ontario
• OSTI ID: 6375377
• Journal Information: Clin. Chem. (Winston-Salem, N.C.); (United States), Journal Name: Clin. Chem. (Winston-Salem, N.C.); (United States) Vol. 33:8; ISSN CLCHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
CARBON 13
CARBON ISOTOPES
CHOLESTEROL
CHROMATOGRAPHY
DEUTERIUM
EVEN-ODD NUCLEI
GAS CHROMATOGRAPHY
HYDROGEN ISOTOPES
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
ISOTOPE DILUTION
ISOTOPES
LIGHT NUCLEI
MASS
MASS SPECTROSCOPY
MEASURING METHODS
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
SEPARATION PROCESSES
SPECTROSCOPY
STABLE ISOTOPES
STEROIDS
STEROLS
TRACER TECHNIQUES