Abstract: The production of traditional rose vinegar is an open, multilateral fermentation process, which is obtained through a series of reactions such as saccharification, alcohol fermentation and acetic acid fermentation. At the right time, rice supplement, sugar supplement or alcohol supplement can complete the fed-batch fermentation, among which alcohol supplement is closer to the end of the reaction which is the simplest way to achieve the expansion of production and efficiency. However, there has been no report on the improvement of the technique of fed-batch fermentation of rose vinegar production. Fermentation of rose vinegar with indica rice in 500 kg cylindrical fermentation container. Under the natural fermentation conditions, fed-batch fermentation was carried out at initial acidity of 1.5, 2.5 and 3.5 g/100mL, respectively. The ratio of vinegar to wine mash is set at 2:1 (v: v) and 1:1 (v: v). Through direct titration with Flynn reagent, alcohol distillation, indicator and potentiometric titration for acid-base titration, UV spectrophotometer, HPLC method, five important indexes including reducing sugar, alcohol content, acidity, color ratio and organic acid were determined. The effects of initial acidity and fed-batch volume on the fermentation of rose vinegar were studied. The results showed that the appropriate fed-batch volume could not only increase the yield of acetic acid fermentation, but also improve the acidity and efficiency of acetic acid fermentation. The color of rose vinegar depends on the feeding time and fed-batch volume. The earlier the vinegar is added, the closer the color is to traditional rose vinegar, but the more the vinegar is added, the greater the influence on the color is. Combining the change of organic acid composition and content with sensory evaluation results, the relationship between the improved fermentation mode and the formation of main flavor substances was discussed to determine the feasibility of fed-batch fermentation of rose vinegar. When the initial acidity was 2.5 g/100mL, the maximum acidity increased to (5.59±0.27)g/100mL(control group (5.19±0.23)g/100mL) after adding the same amount of mash. Compared with the traditional rose vinegar, the acidity content increased significantly (P<0.05). The time of acetic acid fermentation was shortened from 90 days to 78 days, which indicated that fed-batch fermentation could effectively improve fermentation efficiency and save more manpower, material and financial resources. The total amount of nonvolatile organic acids was (27.15±1.11) mg/mL (control group (24.57±0.69) mg/mL). The improvement of this index could make the samples have the characteristics of soft acidity and strong vinegar taste. The color ratio of the final product is 1.8×105 (the traditional process color rate is 1.9×105), which is consistent with that of the traditional rose vinegar. This study has positive significance for the production of rose vinegar to expand production, improve efficiency and realize continuous production throughout the year.