Objective  To compare the differences between flavonoids and synephrine in Aurantii Fructus Immaturus (AFI) with different origins. To provide basis for the identification of the origin and quality control of AFI.

Methods  24 batches of Citrus aurantium L. and 17 batches of Citrus sinensis Osbeck were collected. UPLC method was used, a Waters Acquity UPLC HSS T3 (100 mm×2.1 mm, 1.8 μm) chromatographic column was used, the gradient elution was carried out with 0.1% formic acid-acetonitrile as the mobile phase, the flow rate was 0.3 mL·min^-1, the detection wavelength was 283 nm (0-8 min) and 330 nm (8-11 min), and the injection volume was 1μL. The UPLC fingerprint was established and the similarity evaluation was carried out, the common peaks were identified. With reference to the method for determination of content of AFI in Volume I of Chinese Pharmacopoeia 2020, the content of synephrine in 41 batches of AFI was determined by HPLC.

Results  The UPLC fingerprint analysis method of flavonoids in AFI was established. The similarity of AFI from the same origin was greater than 0.910. The similarity between the two kinds of AFI was only 0.08. Ten common peaks were identified, the peak 4 (naringin), the peak 6 (neohesperidin) and the peak 8 (trifoliate glycoside) of C. aurantium L. were much higher than those of C. sinensis Osbeck. However, the peak 3 (citrulline), peak 5 (hesperidin) and peak 7 (vanillin) of the C. sinensis Osbeck were much higher than those of C. aurantium L. In 17 batches of C. sinensis Osbeck, the highest content of synephrine was 1.700%, the lowest was 0.310%, and the average content was 0.530%. In 24 batches of C. aurantium L., the highest content of synephrine was 0.530%, the lowest was 0.019%, and the average content was 0.290%.

Conclusion  The established UPLC fingerprint analysis method is simple and fast. The fingerprints of the two kinds of AFI were obviously different. It can be used as a reliable method to identify the origin of AFI. The content of synephrine in C. aurantium L. is generally lower than that in C. sinensis Osbeck, and the difference between batches was greater. It is more reasonable to establish the content limit of synephrine for two kinds of AFI.

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