The invention discloses a method of simultaneously determining content of hesperidin, nitidine chloride and psoralen in eight-tradtional-medicine-herb Longzuan particles through HPLC. The chromatographic conditions adopted in the method are as follow: the specification of the chromatographic column Phenomenex Gemini C18 is (250 mm*4.6 mm, 5 micrometers), the mobile phase is acetonitrile, a 0.1% phosphoric acid water solution is used for gradient elution, the flow velocity is 1 ml / min, the detection wavelength is 286 nm and 245 nm (0-30 min: 286 nm, 30-40 min:245 nm), the column temperature is30 DEG C, and the sample size is 5 micrometers. Under the chromatographic conditions, chromatographic peaks are separated well, good linear relations are formed between hesperidin, nitidine chloride and psoralen and the respective peak areas in the concentration ranges of 84.68-508.1 microgram / milliliter,16.84-101.0 microgram / milliliter and 5.504-33.02 microgram / milliliter (r values are 0.99053, 1.00000 and 0.99998 respectively), the average recovery rates are 98.1%, 99.9% and 100.1% respectively, and the RSD values are 1.3%, 2.7% and 2.5% respectively (n=6). The method is simple, rapid, accurate, high in reproducibility and capable of providing a theoretical basis for comprehensively evaluating and controlling the eight-tradtional-medicine-herb Longzuan particles.