Separation And Purification Of Active Ingredients In Fresh Tobacco Leaves

The planting acreage and output of tobacco in China ranks first in the world.After harvesting,about one million tons of fresh tobacco were discared every year in tobacco fields.Fresh tobacco leaves contain a lot of active ingredients with higher potential application value,such as chlorophyll,protein,nicotine,solanesol,and spices.Their reasonable utilization will bring huge economic and social benefits.To this end,this study developed a new process for the comprehensive utilization of active ingredients in fresh tobacco leaves.Fresh tobacco leaves collected from Yunnan Province were crushed and solid-liquid separated to obtain the solid tobacco leaves and liquid.The contents of chlorophyll,nicotine,solanesol and spices in solid tobacco leaves were 0.32%,0.42%,0.21% and 0.12%,respectively.The chlorophyll obtained after double heat reflux were converted into copper sodium chlorophyll after saponification,concentration,extraction,acidification,copper generation,and salt formation,successively.The yield and purity of copper sodium chlorophyll were 82.05% and 93.96%,respectively,and no nicotine was detected in the target product.The nicotine,solanesol and spices were extracted using petroleum ether with a yield of 42.53%,95.62% and 45.00%,and then concentrated to obtain the extract.The solid fraction were used as raw material to produce the bacterial manure after the solid-state fermentation with Bacillus subtilis.The content of protein in liquid was 1.7%.After precipitating at isoelectric point,forty-five percentage of crude protein were obtained.On the one hand,the crude protein was freeze-dried to get the powder,in which the protein was 58.50%,nicotine 0.28%,and water 4.00%.On the other hand,the crude protein is dissolved with 0.1 mol/L NaCl solution and at pH 7.5 for complete dissolution.The protein solution is expected to be purified by anion exchange chromatography.This study has preliminarily established a brand-new process route for the comprehensive utilization of fresh tobacco leaves,achieving the preparation of copper sodium chlorophyllin,nicotine,solanesol,spice extracts,proteins and bacterial manure.This straightforward and effective process are expected to provide technical support for industrial application.