Quantification of endogenous retinoids.
Numerous physiological processes require retinoids, including development, nervous system function, immune responsiveness, proliferation, differentiation, and all aspects of reproduction. Reliable retinoid quantification requires suitable handling and, in some cases, resolution of geometric isomers that have different biological activities. Here we describe procedures for reliable and accurate quantification of retinoids, including detailed descriptions for handling retinoids, preparing standard solutions, collecting samples and harvesting tissues, extracting samples, resolving isomers, and detecting with high sensitivity. Sample-specific strategies are provided for optimizing quantification. Approaches to evaluate assay performance also are provided. Retinoid assays described here for mice also are applicable to other organisms including zebrafish, rat, rabbit, and human and for cells in culture. Retinoid quantification, especially that of retinoic acid, should provide insight into many diseases, including Alzheimer's disease, type 2 diabetes, obesity, and cancer.
Kane
Maureen A
MA
Department of Pharmaceutical Sciences, University of Maryland, Baltimore, MD, USA.
Napoli
Joseph L
JL
D053000:Analytic Sample Preparation Methods; D000818:Animals; D002849:Chromatography, Gas; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D005136:Eye Proteins / Q000032:analysis / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D051379:Mice; D010084:Oxidation-Reduction; D012015:Reference Standards; D015203:Reproducibility of Results; D012172:Retinaldehyde / Q000737:chemistry / Q000302:isolation & purification; D012176:Retinoids / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D013048:Specimen Handling; D013056:Spectrophotometry, Ultraviolet; D013347:Subcellular Fractions / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D014801:Vitamin A / Q000737:chemistry / Q000302:isolation & purification
D005136:Eye Proteins; D012176:Retinoids; D014801:Vitamin A; D012172:Retinaldehyde