钴60辐照对柴胡药材有效成分的影响研究

目的：建立柴胡药材活性成分含量的HPLC测定方法，考察钴60射线辐照前后活性成分含量的变化。方法：采用C₁₈色谱柱（4.6 mm&#215;250 mm，5 μm），流动相为0.1%磷酸水溶液-乙腈，梯度洗脱，流速1.0 mL&#183;min^-1，检测波长210 nm，柱温30℃。分别选择剂量为5、8、10 kGy对柴胡药材进行辐照，比较辐照前后活性成分含量的变化，并进行成组t检验考察其显著情况。结果：柴胡皂苷a、c、d质量浓度分别在0.020~0.806、0.022~0.893及0.025~1.003 mg&#183;mL^-1范围内有良好线性，平均加样回收率分别为99.4%、98.7%和99.8%；未经辐照的6批柴胡药材中柴胡皂苷a、c、d含量范围分别为3.023~3.245、1.034~1.341和5.023~5.298 mg&#183;g^-1；经10 kGy辐照后，上述3个成分的含量范围分别为2.975~3.226、0.823~0.978和4.963~5.136 mg&#183;g^-1；经过5、8、10 kGy辐照后，柴胡药材所含的成分柴胡皂苷a、c、d均有变化，经成组t-检验，在超过5 kGy后，柴胡皂苷c辐照前后含量变化显著（P<0.01）。结论：所建立的柴胡活性成分测定方法可作为柴胡质控方法。在辐照剂量不超过5 kGy时，各成分变化不显著，可为柴胡药材辐照灭菌手段提供参考。

Objective: To establish an HPLC method for determination of effective ingredients in Radix Bupleuri, and to investigate the effect of ⁶⁰Co-γ ray irradiation on the effective components.Methods: The C₁₈ column (4.6 mm&#215;250 mm, 5 μm)was adopted and the mobile phase consisted of 0.1% acetic acid(A)and acetonitrile (B)with a gradient elution at the flow rate of 1.0 mL&#183;min^-1, the wavelength of UV detection was 210 nm, and the column temperature was 30℃.Radix Bupleuri was irradiated at the dose of 5, 8 and 10 kGy.Then contents of the active ingredients before and after irradiation were compared by t-test.Results: The linear range of saikosaponin a, saikosaponin c and saikosaponin d were 0.020-0.806 mg&#183;mL^-1, 0.022-0.893 mg&#183;mL^-1 and 0.025-1.003 mg&#183;mL^-1, respectively, and the average recoveries were 99.4%,98.7% and 99.7%, respectively.The content ranges of saikosaponin a, saikosaponin c and saikosaponin d in 6 non-irradiated Radix Bupleuri were 3.023-3.245 mg&#183;g^-1, 1.034-1.341 mg&#183;g^-1 and 5.023-5.298 mg&#183;g^-1, respectively; while the content ranges of those in the samples irradiated at 10 kGy were 2.975-3.226 mg&#183;g^-1, 0.823-0.978 mg&#183;g^-1 and 4.963-5.136 mg&#183;g^-1, respectively.After irradiation at 5, 8 and 10 kGy, saikosaponin a, saikosaponin c and saikosaponin d in Radix Bupleuri changed.And significant difference was found between contents of saikosaponin c in samples irradiated at 5 kGy.Conclusion: The established method could be used in quality control of Radix Bupleuri. When the radiation was no more than 5 kGy, each active ingredient did not change significantly.The proposed method provided reference for sterilization of Radix Bupleuri.

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