The polybasic acids and higher fatty acids in tobacco were methyl esterified under two derivative conditions: heating-refluxing and placing at room temperature. The reaction mixtures were analyzed with gas chromatography (GC), the results were compared by means of t test. The experimental results revealed that: 1) The results existed significant difference between the two derivative conditions, and refluxing derivation was preferred in the case of precise quantification. 2) In the established method, polybasic acids and higher fatty acids in tobacco were derived into methyl esters by refluxing with sulfuric acid/methanol solution, then analyzed with GC. The recoveries of the method ranged from 90.72% to 98.92% with RSDs below 3%. The derivative method of refluxing features high accuracy and good repeatability, is suitable for determining polybasic acids and higher fatty acids in batches of tobacco samples.%采用加热回流、室温放置两种衍生方法对烟草中多元酸和高级脂肪酸进行甲酯化,并进行气相色谱(GC)分析和t检验.结果表明:①两种衍生化方法的分析结果存在显著性差异.在精确定量时,加热回流是更为合适的衍生化方式.②建立的以硫酸-甲醇溶液为衍生化试剂,加热回流为衍生条件,气相色谱法测定烟叶中多元酸和高级脂肪酸的分析方法,回收率为90.72％～98.92％,RSD小于3％.加热回流的衍生化方法准确度高、重复性好,适合烟叶样品中多元酸和高级脂肪酸的分析.
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