期刊
ANALYTICAL BIOCHEMISTRY
卷 378, 期 1, 页码 71-79出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2008.03.038
关键词
HPLC/UV; retinol; retinyl ester; retinal; retinoids; vitamin A; quantification; mouse; tissue
资金
- NIAAA NIH HHS [R01 AA017927] Funding Source: Medline
- NIA NIH HHS [R01 AG013566-13, AG13566, R01 AG013566] Funding Source: Medline
- NIDDK NIH HHS [R01 DK047839, DK36870, R01 DK036870, F32 DK066924, DK46839, R01 DK047839-15, R01 DK090522, R01 DK036870-22] Funding Source: Medline
We report robust HPLC/UV methods for quantifying retinyl esters (RE), retinol (ROL), and retinal (RAL) applicable to diverse biological samples with lower limits of detection of 0.7, 0.2, and 0.2 pmol, respectively, and linear ranges greater than 3 orders of magnitude. These assays function well with small, complex biological samples (10-20 mg tissue). Coefficients of variation range from 5.9 to 10.0% (intraday) and from 5.9 to 11.0% (interday). Quantification of endogenous RE, ROL, and RAL in mouse serum and tissues (liver, kidney, adipose, muscle, spleen, testis, skin, brain, and brain regions) reveals utility. Ability to discriminate spatial concentrations of ROL and RE is illustrated with C57BL/6 mouse brain loci (hippocampus, cortex, olfactory bulb, thalamus, cerebellum, and striatum). We also developed a method to distinguish isomeric forms of ROL to investigate precursors of retinoic acid. The ROL isomer assay has limits of detection between 3.5 and 4.5 proof and has a linear range and coefficient of variation similar to those of the RCL/RE and RAL assays. The assays described here provide for sensitive and rigorous quantification of endogenous RE, ROL, and RAL to elucidate retinoid homeostasis in disease states such as Alzheimer's disease, type 2 diabetes, obesity, and cancer. (c) 2008 Elsevier Inc. All rights reserved.
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