Abstract: In this study, Streptococcus thermophilus 6063, Lactobacillus bulgaricus 6064 and Bifidobacterium animalis B15 were selected to ferment quinoa yogurt, and the puffed quinoa powder and milk were used as raw materials. Fermentation process was optimized by orthogonal experiment on the basis of single-factor experiments. And the effects of puffed quinoa powder on quality and flavor of yogurt were also evaluated. Results indicated that the optimal fermentation conditions were as follows: Streptococcus thermophilus: Lactobacillus bulgaricus: Bifidobacterium animalis=2:1:2, inoculation amount 2%(v/v), quinoa powder 5%(w/v) and fermentation temperature 41 ℃. Under these conditions, quinoa yogurt had good flavor, and the total viable number of lactic acid bacteria reached 10⁹ CFU/mL. From the beginning of fermentation to the end of storage for 21 days, the titratable acidity of quinoa yogurt was always higher than that of plain yogurt, but the total number of viable lactic acid bacteria was opposite trend. Compared with plain yogurt, the contents of protein, polyphenols and flavonoids in quinoa yogurt increased respectively by 88.67%, 166.67%, and 284.62%. The contents of lactic acid, formic acid, acetic acid and pyruvate in quinoa yogurt were 5.80, 1.42, 0.11 and 0.01 mg/g, respectively. 74 kinds of volatile substances were detected in quinoa yogurt by GC-MS. Compared with pure yogurt, 30 kinds of new volatile substances were discovered. The research results would provide a technology support for further research and new product development of quinoa yogurt, promoting the application and development of quinoa.