An indirect method for the determination of H2S in main stream smoke of cigarette by HPLC with DAD detector was developed.The influence of concectration of zinc acetate on trapping effectiveness of H2S in main stream smoke was investigated.The optimal conditions were using 30 mmol/L zinc acetate-20 mmol/L sodium acetate as trapping reagent system, and 180 mmol/L H2SO4-3 mmol/L N,N-dimethyl-p-phenylenediamine dihydrochloride-10 mmol/L ammonium ferric sulfate as derivation reagent system.The derivative product methylene blue was separated on an Agilent ZORBAX Eclipse XDB-C18(150 mm×4.6 mm,3.5 μm), using a mobile phase composed of methanol-formic acid-water(45∶1∶55) at a flow rate of 0.8 mL/min.DAD wavelength was set at 665 nm.Under the above optimal conditions, the calibration curve was linear in the concentration range of 10.65-532.69 μg/L H2S(0.1-5.0 mg/L for methylene blue) with a correlation coefficient of 0.999 89.The spiked recoveries of S2-were in the range of 96%-98% with relative standard deviations(RSDs) of 2.4%-3.2%.The detection limit(MDL) for H2S in main stream smoke of cigarette was 0.12 μg/cig.The method is simple, rapid and accurate, and could be applied in the determination of H2S in main stream smoke of cigarette.