Abstract: A method for determining the nitrite in flue-cured tobacco with ultraviolet-visible spectrophotometry was established. The factors in extraction process, including extraction decolorization, extraction effect, cleansing, and injection rate in determination were tested. Meantime, the stability, precision and accuracy of this method were investigated, and the nitrite contents in different flue-cured tobacco samples were determined by this method. The results showed that this method was simple in operation, featured good stability, high precision and wide applicability with the recovery from 84.8% to 98.2%, and was suitable for the determination of nitrite in flue-cured tobacco.