Abstract:

A reversed-phase high performance liquid chromatographic (RP-HPLC) method for the determination of carotenoids in flue-cured tobacco leaves was developed. Carotenoids were extracted from flue-cured tobacco leaves by acetone-water (90∶10, v/v) solution containing 0.1% butylated hydroxytoluene (BHT). Plant proteins were eliminated by adding 0.1 g Pb(Ac)2 and by centrifugation (10000 r/min) for 5 min at 4 ℃. Lutein, β-carotene, neoxanthin, violaxanthin and other plant pigments were separated on a reversed-phase C18 column (3.9 mm i.d.×150 mm, 5 μm), with a mobile phase of (A) methanol-isopropyl alcohol (1∶1, v/v) and (B) water using a gradient elution at a flow rate of 0.5 mL/min. The optimum elution gradient was as follows: 0-10 min, 70%A+30%B; 10-17 min, 100%A; 17-30 min, 90%A＋10%B. The recoveries of carotenoids in flue-cured tobacco leaves were 91.77%-97.42%, and relative standard deviations were 3.46%-0.98%. This method was applied to determine carotenoids in flue-cured tobacco leaves during its growth with satisfactory results.

Key words: flue-cured tobacco leaves , carotenoids, reversed-phase high performance liquid chromatography (RP-HPLC)