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目的建立高效液相色谱法测定大鼠血浆中安石榴苷的含量,并用于药代动力学研究。方法血浆样品采用色谱甲醇处理,以大黄素作为内标,Agilent XDB-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为甲醇(A)-0.1%三氟乙酸水溶液(B)梯度洗脱,流速为1.0 ml/min,检测波长为260 nm,柱温为30℃。采用大鼠单剂量静注13 mg/kg的安石榴苷,HPLC法测定安石榴苷的血药浓度,并采用DAS 2.0软件计算药代动力学参数。结果方法学实验结果表明内源性杂质不干扰安石榴苷和内标的测定,线性范围0.02 016~1.00 800 mg/ml。方法精密度、准确度、稳定性和回收率均符合生物样品测定的要求。结论本方法操作简便、灵敏、专属性强,方法学考证符合生物样品测定的要求,并成功用于安石榴苷在大鼠体内的药代动力学研究。
Objective To establish a HPLC method for the determination of pomegranate in rat plasma and to study its pharmacokinetics. Methods The plasma samples were treated with methanol and chromatographed on an Agilent XDB-C18 (250 mm × 4.6 mm, 5 μm) column using emodin as the internal standard. The mobile phase consisted of a gradient of methanol (A) - 0.1% trifluoroacetic acid in water (B) Off, the flow rate of 1.0 ml / min, detection wavelength of 260 nm, the column temperature is 30 ℃. The rat single dose of intravenous injection of pomegranate 13 mg / kg, HPLC determination of punicala blood concentration, and the use of DAS 2.0 software to calculate the pharmacokinetic parameters. Results The results of methodological experiments showed that endogenous impurities did not interfere with the determination of punicalagin and internal standard. The linear range was 0.02 016 ~ 1.00 800 mg / ml. The precision, accuracy, stability and recovery of the method are in line with the requirements of biological samples. Conclusion The method is simple, sensitive and specific. The methodological verification meets the requirements of biological sample determination and has been successfully applied to the pharmacokinetic study of Pomegranate in rats.