The fluorescence spectral characteristic of berberine hydrochloride（BER） was studied in the cucurbit（n=8）uril（Q［8］） aqueous medium.In this medium,Q［8］ and BER formed a supramolecular compound which resulted in the increase of BER fluorescence intensity.A good linear relationship between fluorescence intensity of the system and BER concentration was obtained.Based on this,a novel fluorimetric method was developed for the determination of BER concentration in the cucurbit（n=8）uril medium.Under the optimum conditions,the calibration curve was linear over BER concentration range of 1.90×10-8-1.00×10-5 mol?L-1,with a correlation coefficient（r） of 0.996 1 and a detection limit of 1.90×10-8 mol?L-1.The proposed method was simple,accurate,and could avoid using the toxic solvents and tedious solvent-extraction.And the method was applied in the determination of BER in tablets,serum and urine sample,and the result was in good consistence with those obtained by the pharmacopoeia method.