A method was developed for the determination of conjugated linoleic acid（CLA） in sesame by hollow fiber based two-phase liquid phase microextraction（LPME） coupled with high performance liquid chromatography（HPLC）.For LPME， effects of pH value of extraction solvent，extraction time and stirring rate were investigated，and the optimum experimental conditions were as follows：pH value of extraction solvent（KOH solution）：11.0， extraction time：30 min， stirring rate：1 000 r/min.The qualitative and quantitative analysis of CLA were performed by HPLC method.The proposed method provided a wide linear range with a correlation coefficient（r2） of 0.993，a-detection limit（S/N=3） of 17 μg/L.The relative standard deviation（RSD，n=6） was 4.9% and the enrichment factor of CLA was 12.4. The recoveries of CLA in three kinds of sesame ranged from 81.4% to 94.8%.The method is simple and effective in the determination of CLA in plant，and could also be used in the determination of CLA in the other complex matrix samples such as dairy products，vegetable oils，etc.