The invention provides an analysis method of a nicotine chiral isomer in an electronic cigarette. The method specifically comprises the following steps of (1) sample pretreatment; (2) standard solution preparation; (3) qualitative detection of a sample; and (4) quantitative detection of the sample. According to the analysis method of the nicotine chiral isomer in the electronic cigarette, pretreatment method research and liquid chromatographic condition optimization are carried out on electronic cigarette liquid by employing nicotine as a research object, and a liquid chromatographic analysis method of the nicotine chiral isomer in the electronic cigarette liquid is built, so that the analysis method is good in separation degree and high in sensitivity, and accurate determination of components of the nicotine chiral isomer can be achieved.