The present invention provides a preparation method and application of composite tobacco leaf extract, which uses air-dried tobacco stems, corncobs, and rice bran composite biological materials as immobilized carriers, absorbs microbial aroma-producing strains, incubates, hydrolyzes and ferments composite air-dried tobacco leaf raw materials, and then Combined with the continuous countercurrent ultrasonic extraction technology, the compound tobacco leaf extract is prepared, which has a strong natural tobacco aroma, with rich natural compound aromas such as fresh fruit, wine, milk, barbecue, toasted bread and grilled cheese, etc., irritating reduced, the quality of spices is improved, the purification and separation steps are simple, the process conditions are mild, the extraction rate and production efficiency are improved; it is used in traditional cigarettes and new heat-not-burn cigarettes, and it is coordinated with cigarette aroma and style, increasing smoke concentration and improving satisfaction The taste is rich, the aroma is rich, the irritation is reduced, and the taste is improved.