HPLC-DAD法测定平胃散中非挥发性成分的含量

目的:建立一种利用HPLC-DAD法同时对平胃散中非挥发性成分苍术素、橙皮苷、甘草酸、和厚朴酚、厚朴酚的含量进行测定的方法。方法:采用反相高效液相色谱法,使用GRACE Alltima^TM C₁₈色谱柱(250 mm×4.6 mm,5μm),以0.2%磷酸水(A)-乙腈(B)为流动相,梯度洗脱(0~40 min,10%B→50%B;40~60 min,50%B→95%B),流速1 mL·min^-1,柱温25℃,检测波长235 nm。结果:苍术素未被检测到,橙皮苷、甘草酸、和厚朴酚、厚朴酚进样量分别在4.125~7.612 μg(r=1.000)、3.12~6.24 μg(r=0.9999)、0.305~0.818 μg(r=0.9998)、0.352~1.232 μg(r=1.000)范围内线性关系良好;在精密度和重复性考察中,各测定成分的RSD均小于3%,表明仪器的精密度及方法的重复性良好;稳定性试验表明供试品溶液中各测定成分在24 h内稳定性良好;加样回收试验中橙皮苷、甘草酸、和厚朴酚、厚朴酚平均回收率分别为100.1%、99.98%、101.4%、101.2%,RSD分别为0.17%、1.02%、1.11%、1.24%。橙皮苷、甘草酸、和厚朴酚、厚朴酚在复方中的含量依次为1.248、0.869、0.150、0.104 mg·g^-1。结论:1)苍术素虽为中国药典规定的苍术中含量测定成分,但其在平胃散复方水煎液中并未被检测到,不能作为平胃散复方水煎液质量控制的成分之一;2)利用HPLC-DAD法首次建立一种同时测定平胃散中非挥发性成分橙皮苷、甘草酸、和厚朴酚、厚朴酚含量的方法,该方法简便准确,重复性好,为平胃散质量标准的建立提供参考。

Objective:To develop an HPLC-DAD method for simultaneous determination of atractylodin, hesperidin, glycyrrhizic acid, honokiol, and magnolol in Pingwei San.Methods:The analysis was carried out on a GRACE Alltima^TM C₁₈(250 mm×4.6 mm, 5 μm)column with a mobile phase consisted of 0.2% phosphoric acid(A)and acetonitrile(B)for gradient elution(0-40 min, 10%B→50%B;40-60 min, 50%B→95%B)at the flow rate of 1 mL·min^-1.The column temperature was 25℃ and the detection was set at 235 nm.Results:Atractylodin was undetectable;the liner ranges of hesperidin, glycyrrhizic acid, honokiol and magnolol were 4.125-7.612 μg(r=1.000), 3.12-6.24 μg(r=0.999 9), 0.305-0.818 μg(r=0.999 8)and 0.352-1.232 μg(r=1.000);the study of precision and repeatability showed that the precision of the instrument and the method of repeatability were favorable, and the RSD values of determination were less than 3%.The stability test showed that the determination of the components in sample solution was stable within 24 h;the recoveries were 100.1%, 99.98%, 101.4%、101.2% and the RSDs were 0.17%, 1.02%, 1.11% and 1.24%, respectively.The contents of hesperidin, glycyrrhizic acid, honokiol and magnolol were 1.248, 0.869, 0.093 and 0.104 mg·g^-1.Conclusion:Although atractylodin is specified by Pharmacopoeia of the People's Republic of China 2010 content determination of constituents of Rhizoma Atractylodis, it has not been detected in Tangji of Pingwei San.So it cannot be used as an indicator of quality control for Tangji of Pingwei San.The established method for determination of hesperidin, glycyrrhizic acid, honokiol, magnolol in Pingwei San is simple and exactly, which can be used to control the quality for Pingwei San.