Abstract: To meet the need of the research on isotope effects of mixed hydrogen isotope gas, an analysis method of mixed hydrogen isotope composition was set up through introducing the molecule ionization quotient on MAT253 mass spectrometer. Some effect factors to quotient were examined and the method is used to determine 6 samples of different deuterium concentration. The results show that this method is effective for P2D2 mixed gas and it’s relative standard deviation is more low then 0.3%.