Abstract
Extraction is most commonly used sample preparation technique in quantitative determination of all-trans-lutein in spinach. In our study RP-HPLC was applied to identify the constituents of spinach extract and to quantify all-trans-lutein content with the mobile phase consisted of acetone and water. Due to sensitivity of all-trans-lutein, we compared effects of different extraction conditions; air and nitrogen atmosphere and ethanol, ethanol with added 0.1% TBHQ and ethanol with added 0.1% BHT as solvents. Results for the all-trans-lutein content in the spinach samples and spinach samples with standard addition obtained in air and in nitrogen confirmed the induced degradation of all-trans-lutein in oxygen from the air. Ethanol containing synthetic antioxidant TBHQ under nitrogen atmosphere gave the highest yield of extraction and recovery of all-trans-lutein from spinach.
Highlights
IntroductionCarotenoids are tetraterpenes (composed from eight isoprene units) which are synthesized in bacteria, algae, fungi and green plants [1] where they form lipid-soluble yellow, orange, and red pigments [2]
Carotenoids are tetraterpenes which are synthesized in bacteria, algae, fungi and green plants [1] where they form lipid-soluble yellow, orange, and red pigments [2]
In our study RP-HPLC was applied to identify the constituents of spinach extract and to quantify all-translutein content with the mobile phase consisted of acetone and water
Summary
Carotenoids are tetraterpenes (composed from eight isoprene units) which are synthesized in bacteria, algae, fungi and green plants [1] where they form lipid-soluble yellow, orange, and red pigments [2]. Over 700 known carotenoids [3] are divided into oxygenated xanthophylls such as lutein, zeaxanthin, violaxanthin, neoxanthin and hydrocarbon carotenes such as β-carotene, α-carotene and lycopene. Because of their structure (several possible isomers), their instability, and so forth their extraction, isolation, identification and determination represent a considerable challenge. The most prevalent carotenoids in human serum are α-carotene, β-carotene, lycopene, lutein, zeaxanthin, and β-cryptoxanthin [2]. Since TBHQ and BHT are cheap and commercially available we studied their effect on extraction efficiency of all-trans-lutein from the spinach in air and under nitrogen atmosphere. The first and principal aim of present study was to develop a simple and the most efficient analytical procedure for extraction of all-translutein from spinach and separation method for carotenoids which could be applied in other studies
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