The invention provides establishment of the liquid chromatographic retention index and application of establishment to a compound qualitative aspect. Establishment specifically includes the following steps that firstly, a mixed solution of a retention index marker is prepared; secondly, under the standard chromatographic condition, LC-MS/MS detects the marker and determines the retention index value of the marker; thirdly, fresh tobacco leaves are quick-frozen with liquid nitrogen, freeze-dried and smashed; fourthly, the smashed sample is added into an extraction solvent and subjected to low-temperature ultrasonic extraction; fifthly, refrigerated centrifugation is conducted, and supernate is obtained and filtered; sixthly, LC-MS/MS detects the supermate and conducts qualitative analysis on a compound contained in the solution. Retention time of compound components in complex sample solutions can be corrected through the retention index marker, and retention time change caused by mobile phases, pressure or use of chromatographic columns different in batch number is avoided. Compared with the prior, multiple times of experiments under the same condition are not needed, the process is simple, cost is low, and qualitative accuracy and uniqueness can be greatly improved.