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Determination of blood concentration of active component in Bupleurum Injection |
XIE Huan TAN Ruyue DENG Jingjing |
School of Pharmacy, Chengdu Medical College, Sichuan Province, Chengdu 610500, China |
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Abstract Objective To establish high performance liquid chromatography (HPLC) for the determination of saikosaponin d of Bupleurum Injection in rat plasma. Methods Six SD male rats were intraperitoneally injected with Bupleurum Injection of 5 mL/kg, and the blood was collected from the tail vein at different time points. The blood samples were treated with ethyl acetate precipitated protein method, and the blood concentration of saikosaponin d was determined by HPLC. The column Swell chromplus-C18 (250 mm×4.6 mm,5 μm) was used as the column. The mobile phase was water-acetonitrile (gradient elute), the detection wavelength was 204 nm, the column temperature was 30℃ and with the flow rate of 1.0 mL/min. Results The linear range of saikosaponin d in rat plasma was 0.1-2 mg/mL (r = 0.9955). The accuracy recovery rates of low, medium and high concentrations were 92.95%, 105.52% and 97.86%, respectively. The RSD values of intra-day and intra-day precision were all < 6%, and the detection limit was 2 μg/mL. Plasma samples with low, medium and high concentrations were kept stable for 12 h at room temperature, frozen at -20℃ for 10 d, and repeatedly frozen and thawed 3 times. Conclusion The method is accurate, specific, and suitable for the determination of saikosaponin d of Bupleurum Injection in rat plasma.
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