Absorbance changes of carotenoids in different solvents

Carotenoids are typically measured in tissues with the high performance liquid chromatography (HPLC) and quantitation is usually done by calibrating with stock solutions in solvents. Four carotenoids including lutein, zeaxanthin, lycopene and beta-carotene were dissolved in hexane and methanol respectively, and their absorbance characteristeris were compared. Lutein shows absorbance spectra that are almost independent of solvents at various concentrations. Spectra of zeaxanthin, lycopene and beta-carotene were found to be more solvent-dependent. The absorbance of zeaxanthin at lambda(max) is about similar to 2 times larger in methanol than in hexane at the higher concentrations, and increased non-linearly with increasing concentration in hexane. The absorbance of lycopene at lambda(max) in hexane is similar to 4 fold larger than in methanol, but the absorbance of the methanol sample can be recovered by re-extracting this sample in hexane. The absorbance of beta-carotene in hexane is larger than in methanol, and increased linearly with increasing concentration. But beta-carotene showed a non-linear concentration effect in methanol. There are very small variations in lambda(max) for all four carotenoids between hexane and methanol, due to differences in molar extinction coefficients. The non-linear concentration effects for these carotenoids are probably due to differences in solubility leading to the formation of microcrystals. Thus, care should be taken with quantitation of tissue carotenoid values, when they depend on measurement of concentrations in stock solutions. (C) 1997 Elsevier Science Inc.