Objective: To establish an HPLC method for simultaneous determination of loganic acid, gentiopicroside, hesperidin, paeonol, aloe-emodin, rhein, emodin, chrysophanol and physcion in Tongshu Koushuang tablets (Rhei Radix et Rhizoma, Aurantii Fructus Immaturus and Gentianae Macrophyllae Radix, etc.). Methods: The samples were extracted by refluxing with methanol and the separation was performed on a Thermo Acclaim-C18(250 mm×4.6 mm, 5 μm) column at 30 ℃, with mobile phase consisting of 0.15% phosphoric acid solution and acetonitrile with a gradient elution at a flow rate of 1.0 mL·min-1. The injection volume was 10 μL, and the detection wavelength was set at 236 nm (detecting loganic acid), 280 nm (detecting gentiopicroside, hesperidin and paeonol) and 254 nm (detecting aloe-emodin, rhein, emodin, chrysophanol and physcion). Results: Nine constituents (loganic acid, gentiopicroside, hesperidin, paeonol, aloe-emodin, rhein, emodin, chrysophanol and physcion) were in good linearity within their own ranges (r were 0.999 1-0.999 9), and the average recoveries were 99.6% (RSD=1.9%), 101.1% (RSD=2.0%), 103.0% (RSD=1.5%), 101.6% (RSD=0.51%), 99.8% (RSD=1.4%), 100.9% (RSD=1.4%), 100.8% (RSD=1.8%), 102.7% (RSD=1.2%) and 102.9% (RSD=0.86%), respectively. The content ranges of nine constituents were 1.871-3.517 mg·g-1, 6.274-12.55 mg·g-1, 2.140-3.946 mg·g-1, 1.393-2.018 mg·g-1, 0.394 8-0.807 2 mg·g-1, 0.657 5-1.246 mg·g-1, 0.484 7-0.915 8 mg·g-1, 0.839 8-1.429 mg·g-1 and 0.327 3-0.547 9 mg·g-1, respectively. Conclusion: This simple and accurate method can be used for the quality control and evaluation of Tongshu Koushuang tablets.
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