A ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for the determination of female-steroid hormones in brown sugar. Brown sugar samples were dissolved in water, and extracted by acetonitrile, then cleaned-up using an HLB column. The analytes were separated on a ACQUITY UPLC HSS T3 column with acetonitrile-water (containing 2 mmol/L ammonium acetate) as mobile phase by gradient elution. The mass spectrometer was detected in multiple reaction monitoring mode(MRM) using positive-ion and negative-ion electrospray ionizations(ESI), respectively. The quantitative analysis was carried out by external standard method. The results showed that the calibration curves had good linearity for 9 female-steroid hormones in the tested concentration ranges, with correlation coefficients of more than 0.99. The average recoveries at three spiked levels were 92.2%~113.2%, and the precisions expressed as relative standard deviations (RSDs) were 1.6%~6.8%. The limits of detection (LOD) were 0.2~1.0 μg/kg. The method proved to be accurate, rapid, sensitive with repeatability and suitable for rapid detection of female hormone in brown sugar.