Abstract: The isotope dilution mass spectrometry can avoid the matrix effect and the influence of analysis conditions, which has quantitative analysis and the accuracy of results in absolute precision special advantage, so it is widely applied in the field of quantitative analysis of biomolecules. The stable isotopic labeled peptides are widely used as an important internal standard reagent in the quantitative of proteomicsand food allergenic detection. The development of isotopic dilution mass spectrometry will further promote the research and application of stable isotope labeling technology in new drug development, clinical pathology, proteomics and food safety. With the rapid development of stable isotope labeling technology in recent years, the synthesis of stable isotope labeling peptides was summarized briefly from two aspects of biosynthesis method and chemical method. Methods of marker of cell metabolism, higher eukaryotes tag, enzyme catalytic notation, acetylation, labeled amino acid condensation were discussed in details. The application of stable isotopic marker polypeptide in the detection of dairy products, quantitative protein, protein sequencing and drug metabolism and food allergens were introduced.