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目的:建立HPLC法测定家兔血清中穗花杉双黄酮含量的方法。方法:色谱柱:Cosmosil 5C_(18)-MS-Ⅱwaters(250 mm×4.6 mm,5μm),以乙腈为流动相A,水溶液(含2%的四氢呋喃,0.1%的三氟乙酸)为流动相B进行梯度洗脱,pH为2.2,流速:1.0 ml·min~(-1),柱温:30℃,检测波长为335 nm。结果:穗花杉双黄酮在血清中的线性范围为0.051 2～2.05μg·ml~(-1)(r=0.999 5,n=5);低、中、高3个浓度的相对回收率分别为(83.98±0.98)%、(96.24±1.22)%、(100.50±1.38)%,RSD≤1.93%(n=6)。结论:该方法可为穗花杉双黄酮在家兔体内的药物动力学研究提供参考。
Objective: To establish a HPLC method for the determination of biflavone content in rabbit serum. METHODS: The chromatographic column consisted of Cosmosil 5C_ (18) -MS-Ⅱwaters (250 mm × 4.6 mm, 5 μm) using acetonitrile as mobile phase A and aqueous solution containing 2% tetrahydrofuran and 0.1% trifluoroacetic acid as mobile phase B The gradient elution was carried out at a pH of 2.2. The flow rate was 1.0 ml · min -1. The column temperature was 30 ℃ and the detection wavelength was at 335 nm. Results: The linear range of biflavone was 0.051 2 ~ 2.05μg · ml -1 (r = 0.999 5, n = 5). The relative recoveries of low, medium and high concentrations were (83.98 ± 0.98)%, (96.24 ± 1.22)%, (100.50 ± 1.38)%, RSD≤1.93% (n = 6). Conclusion: This method can provide a reference for the pharmacokinetics of Fraxinus mandshurica flavonoids in rabbits.