Study On Extraction, Separation And Purification, Oxidative Degradation, And Their Degradations Application In Flavoring

The carotenoid is one of the most important terebene compounds in tobacco, it not only has direct impact on appearance quality of the tobacco leaves, but also its degradations are closely related to internal quality and fragrance amount. This article studied the best technological conditions for extracting, separation and purification of the carotenoids from tobacco leaves. oxidative degradation of the carotenoids and their degradations application in cigarettes flavoring. Study result as follows:1. Carotenoids were extracted by ultrasonic.The influence of different organic solvent, ultrasonic power, ratio of solid to liquid, extracting temperature and time on extraction of carotenoids was studied. At last, based on the orthogonal experiment, determined the optimal extraction condition: ultrasonic power 100 W, the ratio of solid to liquid 1:25 (g/mL), extracting 30 min under the temperature of 30℃, with petroleum ether-acetone (1:1, v/v).the resulting extraction has highest absorption spectrum value. Some large-scale experiments were conducted based on condition above, the absorption value of carotenoids increased with experimental scale amplified.2. The carotenoids compound of tobacco leaves were separated and purified by thin layer chromatography (TCL) and column chromatography. According to the results of TCL, Petroleum ether -acetone (8:2, v/v) was used as a developer, two kinds of main ingredient in tobacco leaves were determined, the Rf were 0.916, 0.242 respectively. The separation condition of column chromatography was: gradient washed by Petroleum ether–acetone, the spotⅠwas quickly washed by Petroleum ether -acetone (9:1, v/v), he spotⅡwas washed by Petroleum ether -acetone (7:3, v/v),the carotenoids were separated based on the condition above and the results suggested there were two compounds in tobacco leaves, which were consistent with TCL.By color reaction, the mensuration of Rf, and the mensuration of UV-vis, high performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR), at the same time, with the relevant results of standard substances from UV-vis, HPLC and NMR, we can give a qualitative conclusion that the two kinds of main ingredients in tobacco leaves areβ-carotene and lutein respectively.3. Chemistry method was used to oxidative degradation ofβ-carotene and lutein, and the products of degradation were analysised by GC/MS. The results indicated that theβ-carotene degradated 17 kinds of compounds under the effection of H2O2, by using 95% ethanol as solvent, among them, the contents of dihydroactionlide andβ-violet alcohol were relative higher, which reached 40.076% and 21.084%. In the oxidation of H2O2, AgNO3 and KMnO4, the lutein generated 6, 12 and 17 kinds of compounds respectively, common components had dihydroactionlide, 3-oxygen violet alcohol, isophorone and so on. these known produces of degradation can be found equally in natural degradation of carotenoid in tobacco leaves, among them, the produces of degradation such as dihydroactionlide, beta-Ional, isophorone and damascone are the important tobacco flavors.4. the degradation products ofβ-carotene and lutein were applied in unblended cut tobacco and cut filler, the results indicated that adding the suitable amount of degradation products ofβ-carotene and lutein could efficiently improve smoking quality of the cigarettes, increase quantity of tobacco aroma, reduce offensive odor and irritation, amend aftertaste, the rational consumption of degradation products ofβ-carotene and lutein in tobacco flavoring was 1‰.