草乌药材标准修订研究

Revision study on quality standard of Radix Aconiti Kusnezoffii

目的:修订并提高毒性药材草乌的质量控制标准。方法:采用以乌头碱、次乌头碱和新乌头碱对照品为对照的薄层色谱法对草乌药材进行鉴别;采用高效液相色谱法测定其中双酯型生物碱乌头碱、次乌头碱和新乌头碱的含量,色谱柱为Phe-nomenex C18(5μm,4.6mm×250mm),流动相A为乙腈-四氢呋喃(25:15),流动相B为0.1mol.L-1醋酸铵(每1000mL加0.5mL冰醋酸),梯度洗脱[0～48min,A-B(15:85)→(26:74);48～49min,A-B(26:74)→(35:65);49～58min,A-B(35:65)],流速0.8mL.min-1,检测波长235nm,柱温25℃。结果:草乌药材薄层色谱斑点清晰,分离度好。乌头碱、次乌头碱、新乌头碱液相色谱峰分离良好;各成分浓度与HPLC峰面积呈良好的线性关系,线性范围分别为0.0010～0.52μg(r=0.9999),0.0018～0.90μg(r=0.9999),0.0020～2.02μg(r=0.9999);平均回收率(n=9)分别为98.9%,100.8%,96.2%。结论:本研究建立的草乌药材的薄层色谱鉴别法和HPLC含量测定方法专属、准确,耐用性良好,可有效控制草乌药材质量,提高了毒性药材草乌的检测标准。

Objective：To revise and improve the quality standard of toxic Chinese medicinal herb Radix Aconiti Kusnezoffii.Methods：Three alkaloids（aconitine,hypaconitine and mesaconitine） in Radix Aconiti Kusnezoffii were identified and determined by new TLC and HPLC methods.The separation was performed on a Phenomenex C18 analysis column（5 μm,4.6 mm×250 mm）with column temperature of 25 ℃,the mobile phase consisted of acetonitrile-tetrahydrofuran（25：15,phase A）,0.1 mol·L-1 ammonium acetate（per 1000 mL contains 0.5 mL glacial acetic acid,phase B） with gradient elution[0-48 min,A-B（15：85）→（26：74）;48-49 min,A-B（26：74）→（35：65）;49-58 min,A-B（35：65）]at the flow rate of 0.8 mL·min-1,and detector at 235 nm.Results：Clear spots were obtained with good separation in TLC identification of Radix Aconiti Kusnezoffii.The calibration curves were linear in the range of 0.0010-0.52 μg（r=0.9999） for aconitine,0.0018-0.90 μg（r=0.9999） for hypaconitine,0.0020-2.02 μg（r=0.9999） for mesaconitine;The average recoveries（n=9） were 98.9%,100.8%,96.2%,respectively.Conclusion：These new TLC and HPLC methods established in this paper are sensitive,accurate,and suitable for the quality control of Radix Aconiti Kusnezoffii.