Quantitation of biological retinoids by high-pressure liquid chromatography: primary internal standardization using tritiated retinoids

Journal Article · Sat Feb 15 00:00:00 EST 1986 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(86)90055-2· OSTI ID:5698244

Cullum, M E; Zile, M H

A single method is described for quantitation of 14 retinoids found in biological material. The method consists of reversed-phase HPLC, internal standardization, and carrier extraction procedures with three synthetic retinoids. Primary standardization of HPLC uv detector is achieved using tritiated all-trans-retinoic acid, all-trans-retinol, all-trans-retinyl palmitate, and all-trans-retinyl acetate. Extraction methods are standardized by correlating the uv absorbance of retinoids at 340 nm with radioactivity of tritiated retinoids of known specific activity. Quantitation of 10 pg of tritiated or 5 ng of nonradioactive retinoid per 0.1 g sample in a polarity range from 4-oxo-retinoic acid to retinyl stearate can be achieved in a single, 50-min chromatographic run. A single HPLC pump, a C/sub 18/ reversed-phased analytical column, a multistep three-solvent gradient, and inexpensive solvents based on methanol, water, and chloroform comprise this cost-effective chromatographic system. Our primary standardization method allows investigators employing different procedures to compare results between laboratories by standardizing the HPLC uv detector with commercially available tritiated retinoids. With this method we were able to quantitate nanomolar amounts of endogenous retinoic acids and retinyl esters, that HPLC uv only conditions usually would not detect in the circulation and liver of rats under physiological conditions.

• Research Organization: Michigan State Univ., East Lansing
• OSTI ID: 5698244
• Journal Information: Anal. Biochem.; (United States), Vol. 153:1
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
BIOLOGICAL MATERIALS
LIQUID COLUMN CHROMATOGRAPHY
QUANTITATIVE CHEMICAL ANALYSIS
TRACER TECHNIQUES
STANDARDIZATION
LIVER
RATS
RETINOIC ACID
SOLVENT EXTRACTION
TRITIUM COMPOUNDS
ULTRAVIOLET SPECTRA
VITAMIN A
ANIMALS
BODY
CARBOXYLIC ACID ESTERS
CHEMICAL ANALYSIS
CHROMATOGRAPHY
DIGESTIVE SYSTEM
ESTERS
EXTRACTION
GLANDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MATERIALS
ORGANIC COMPOUNDS
ORGANS
RODENTS
SEPARATION PROCESSES
SPECTRA
VERTEBRATES
VITAMINS
400102* - Chemical & Spectral Procedures
550201 - Biochemistry- Tracer Techniques