[Determination of carotenoids in flue-cured tobacco leaves during its growth by reversed-phase high performance liquid chromatography].
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 degrees C. Lutein, beta-carotene, neoxanthin, violaxanthin and other plant pigments were separated on a reversed-phase C18 column (3.9 mm i.d. x 150 mm, 5 microm), 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.
Liu
Guoshun
G
National Tobacco Cultivation, Physiology and Biochemistry Research Center, Henan Agricultural University, Zhengzhou 450002, China.
Wei
Fengjie
F
Wang
Fang
F
Li
Yajuan
Y
Guo
Qiaoyan
Q
Huang
Xiaoshu
X
D002338:Carotenoids / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D014975:Lutein / Q000032:analysis; D014026:Nicotiana / Q000737:chemistry*; D024341:Xanthophylls / Q000032:analysis; D019207:beta Carotene / Q000032:analysis*
D024341:Xanthophylls; D019207:beta Carotene; D002338:Carotenoids; C005613:violaxanthin; C011947:neoxanthin; D014975:Lutein