炮制对天南星毒性成分草酸钙针晶及凝集素蛋白含量的影响

Effect of Arisaematis Rhizoma Processed on the Contents of Lectin Protein and Calcium Oxalate Crystal as Toxic Components

    摘要
  • 摘要:
      目的  探究天南星炮制前后内源性毒性成分的含量变化, 建立天南星毒性控制指标检测方法。
      方法  采用HPLC法对35批天南星炮制前后草酸钙含量进行测定, 色谱柱为Megres C18(250 mm×4.6 mm, 5 μm), 流动相为0.5%磷酸水溶液, 流速0.8 mL · min-1, 检测波长220 nm, 柱温25 ℃; 采用Western blot法对32批天南星炮制前后凝集素蛋白进行检测。
      结果  各批次生天南星草酸钙含量为0.78%~2.33%, 平均含量为1.43%;各批次制天南星草酸钙含量为0.29%~0.98%, 平均含量为0.60%;炮制后草酸钙含量的降幅为25.6%~78.3%。各批次天南星炮制后均未检测到凝集素蛋白条带。
      结论  天南星经过复制法炮制, 可显著降低所含内源性毒性物质草酸钙及凝集素蛋白的含量。建立的草酸钙及凝集素蛋白检测方法可用于天南星毒性成分检测, 以凝集素蛋白限度作为天南星毒性控制指标更为稳定可靠, 可为天南星毒性控制指标的建立提供参考。

     

    Abstract:
      OBJECTIVE  The present study was aimed to investigate the effect of Arisaematis Rhizoma processed on the contents of endogenic toxic components and to establish the detection method.
      METHODS  HPLC was applied to determinate the content of calcium oxalate. The separation was developed on Megres C18 column (250 mm×4.6 mm, 5 μm) by gradient elution with water containing 0.5% phosphoric acid as the moble phase at the flow rate of 0.8 mL·min-1, the column temperature of 25 ℃, and the detection wavelength of 220 nm. Western blot was applied to detect the content of lectin protein.
      RESULTS  The contents of calcium oxalate of all batches of raw Arisaematis Rhizoma was 0.78%-2.33%, and average content was 1.43%;The contents of calcium oxalate of all batches of processed Arisaematis Rhizoma were 0.29%-0.98%, and average content was 0.60%; After processing, the decreasing amplitude of calcium oxalate were 25.6%-78.3%. No lectin protein was detected in all batches of processed Arisaematis Rhizoma.
      CONCLUSION  The contents of toxic components, lectin protein and calcium oxalate, were decreased in Arisaematis Rhizoma after processed. The detection method of lectin protein and calcium oxalate can be used to detect toxic components of multi-batch Arisaematis Rhizoma, and can provide reference for the establishment of toxicity control index of Arisaematis Rhizoma. It is stable and reliable that consider the limit of lectin protein as toxicity control index of Arisaematis Rhizoma.

     

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