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Extraction process for Q-markers from Rosa roxburghii using response surface methodology was optimized and HPLC method was established for simultaneous determination of their content. The extraction process of Q-marker from the root of Rosa roxburghii Tratt was optimized by single factor experiments and response surface methodology, and the four Q-markers (triterpenoids) in different parts (root, stem, leaf and fruit) of Rosa roxburghii Tratt were analyzed. Results showed that, through single factor experiments, the important factors affecting the yield of Q-markers were determined as solvent dosage, ethanol concentration, extraction time and extraction times. Response surface methodology was used to optimize the extraction process of Q-markers from Rosa roxburghii. The optimal extraction conditions were as follows: The solvent dosage was 1:25 g/mL, 70% ethanol was added, the extraction time was 30 min, and then the material was refluxed for 2 h. The yield of four Q-markers from the Rosa roxburghii leaves was 2.64%, including 0.99% of Kaji-ichigoside F1, 0.72% mg/g of rosamultin, 0.43% of rosolic acid and 0.51% of 2α,19α-dihydroxyursolic acid. The total content of Q-markers in different parts of Rosa roxburghii Tratt was in the order of leaves, roots, stems and fruits. This study would provide scientific reference for quality evaluation of Rosa roxburghii and development and utilization of total triterpenoids.