Objective To compare the quality of Rheum palmatum L. in different areas and ages in Gansu Province by comparing five components in Rheum palmatum L. through fingerprints and QAMS (quantitative analysis of multi-components) methods; To determine the most suitable growing areas and the best development areas in Gansu Province. Methods The water content, total ashes, and water-soluble extract content in 15 batches of Rheum palmatum L. from different areas and ages in Gansu Province were determined according to the approaches listed in the Chinese Pharmacopeia. The contents of five index components were determined through HPLC and the quality of 15 batches of Rheum palmatum L were evaluated by fingerprint and QAMS method. Results The results from 15 batches of Rheum palmatum L. showed that the water content was less than 15％, total ashes less than 10％, and water-soluble extract content less than 25％, conforming to Pharmacopoeia. There were 23 common peaks in the fingerprints of 15 batches of Rheum palmatum L.. Five of them were identified as aloe-emodin, rhein, emodin, chrysophanol and physcion, which showed good linear relationship in the range of 0.0122–0.7344 μg (r=0.9999), 0.00714–5.7120 μg (r=0.9999), 0.0088–0.7040 μg (r=1.0000), 0.1224–3.6720 μg (r=0.9999) and 0.0148–5.9200 μg (r=0.9997), respectively. There was no significant difference between the calculated value and the measured value by using the relative correction factor (RCF), and the reproducibility of RCF was good. The quality of Rheum palmatum L. from different areas and ages in Gansu Province significantly differed (P<0.05). Two year old Rheum palmatum L. from Pingxiang village of Li county and Rushu village of Tanchang county had the best quality, and that of three years old was better than two years old from Tanchang county. Conclusion The established fingerprints and QAMS method is accurate, feasible, and can be used for the quality comparison of Rheum palmatum L. from different areas and ages in Gansu Province. Li County and Tanchang County areas in Gansu Province can be used as suitable planting areas and development industry. Therefore, it is recommended to select more than two yeas old Rheum palmatum L. from Tanchang County and Li County in Gansu Province for medicinal application.