Pyrophosphate can seriously affect human health. Therefore， it is necessary to control the content of pyrophosphate in food. In the acid medium， copper ion (Cu2+) can effectively catalyze the redox reaction between hydrogen peroxide and ninhydrin， leading to produce a strong fluorescence. When a small amount of pyrophosphate was added into the hydrogen peroxide-ninhydrin system， the above-mentioned fluorescence was obviously reduced. Based on this observation， a fluorescence strategy for the quantitative analysis of pyrophosphate was developed. Several experiment parameters， such as hydrogen peroxide concentration， ninhydrin concentration， Cu2+ concentration， reaction time and reaction temperature were optimized to investigate the effect of these factors on the fluorescence intensity. The result showed that， the optimized experimental conditions was set as follows: 3.0×10-5 mol/L of hydrogen peroxide， 2.0×10-3 mol/L of ninhydrin 6.0×10-5 mol/L of copper ion， 40 min of reaction time and reaction temperature at 30 ℃. Under the conditions，the relative fluorescence intensity displayed a good linear relationship with the logarithm value of pyrophosphate concentration in the range of 3.0×10-7-8.0 ×10-6 mol/L. And the detection limit and the relative standard deviation were 3.2×10-8 mol/L and 4.9% (C=1.0×10-6 mol/L， n=11)， respectively. The content of pyrophosphate in the shelled shrimp and hairtail samples were quantified by the proposed method， and the average recoveries were between 93.00% and 97.80%. This result showed that the developed method can be applied in the accurate detection of pyrophosphate content in the practical food samples.