固相萃取-反相高效液相色谱法测定大鼠血浆中羟基积雪草苷的浓度

Determination of madecassoside in SD rats plasma by solid-phase extraction RP-HPLC

目的:建立大鼠血浆中羟基积雪草苷测定的高效液相色谱法。方法:固相萃取法对样品进行纯化处理后,选用Welchrom-C18柱(250mm×4.6mm,5μm),水-乙腈(24∶76)为流动相测定药物含量;流速:1.0ml/min;紫外检测波长:225nm;柱温:30℃;内标:葫芦素B。结果:在此色谱条件下,羟基积雪草苷和内标葫芦素B的保留时间分别为5.11min和8.14min。线性范围为5μg/ml～1.5mg/ml(r=0.9998),最低检测浓度为1μg/ml。方法回收率在90.21%～95.65%,提取回收率在80.65%～87.04%。日内与日间精密度的RSD在1.31%～3.07%(n=5)。结论:该检测方法简便可靠,选择性和重现性好,符合常规生物样品检验的要求,可用于羟基积雪草苷在大鼠体内的药代动力学研究。

Objective：To establish a method for madecassoside determination by solid-phase extraction RP-HPLC in SD rats plasma. Methods：The sample was cleaned with an Strata-X solid-phase extraction . HPLC conditions were as follows：Welchrom-C18 column （250 mm×4.6 mm,5 μm）,water-acetonitrile（24∶76）as mobile phase（flowrate：1.0 ml/min）,cucurbitacin B as internal standard, detection wavelength of 225 nm,and column temperature of 30℃. Results：In this situation,the remaining times of the madecassoside and cucurbitacin B were 5.11 min and 8.14 min,respectively. The linear range of the method was 5 μg/ml～1.5 mg/ml （r=0.999 8）.The low limit of detection was 1 μg/ml. The method recoveries and the extraction recoveries were 90.21%～95.65% and 80.65%～87.04%,respectively. The within-day RSD and between-day RSD were between 1.31%～3.07%（n=5）. Conclusion：Themethod is simple,convenient,accurate, reproducible,and suitable for studying the pharmacokinetics of madecassoside in SD rats .