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Nitrogen content matters in tobacco – alkaloids, proteins, amino acids, nitrates, ammonia and other nitrogen bases have big importance for tobacco quality and tobacco industry. The quantity of nitrogen content matters in tobacco raw is determined like total nitrogen. One of important factor for these matters accumulation in tobacco is nitrogen fertilization. Burley type tobacco has more nitrogen content matters in comparison to other tobacco types produced in our country – Oriental and Virginia. The comparative investigation is made of main alkaloid content – nicotine, nitrate content and total quantity of nitrogen content matters – total nitrogen in Burley tobacco varieties. It is determined the influence of variety, fertilization norm and harvesting belt on quantity of nitrogen content matters in investigated tobacco. The factors knowledge influenced on these matters accumulation give ability for regulation its content in tobacco.