摘要
建立了采用反相高效液相色谱测定烤烟叶片中类胡萝卜素的方法。烤烟叶片先用含0.1%丁基羟基甲苯(BHT)的90%丙酮水溶液萃取,然后加入0.1g醋酸铅,于4℃下以10000r/min离心5min以去除蛋白质。色谱柱为C18反相柱(3.9mmi.d.×150mm,5μm)。流动相:A,甲醇-异丙醇(体积比为1:1);B,超纯水。洗脱程序:0~10min,70%A+30%B;10~17min,100%A;17~30min(90%A+10%B)。流速:0.5mL/min。进样量:10μL。检测波长:450nm。该方法简化了样品的前处理过程,4种类胡萝卜素物质的加标回收率为91.77%~97.42%,相对标准偏差为3.46%~0.98%。用该方法研究了烤烟发育过程中类胡萝卜素含量的变化规律,获得了与文献较为一致的结果。
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, 5μm), 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 3.45% - 0.98%. This method was applied to determine carotenoids in flue-cured tobacco leaves during its growth with satisfactory results.
出处
《色谱》
CAS
CSCD
北大核心
2006年第2期161-163,共3页
Chinese Journal of Chromatography
基金
国家烟草专卖局项目(No.110200201005)
关键词
反相高效液相色谱法
类胡萝卜素
烤烟叶片
reversed-phase high performance liquid chromatography (RP-HPLC)
carotenoids
flue-cured tobacco leaves