佛波双酯对照品的制备研究

Study on preparation of reference products of phorbol diesters

目的研究佛波双酯(TPA)对照品的制备方法及质量标准,为佛波双酯原料和相关制剂的质量控制提供依据。方法利用硅胶柱制备色谱和反相制备色谱对巴豆油的甲醇-水提取物进行分离纯化,分别以高效液相色谱法(HPLC)和气相色谱对佛波双酯对照品进行纯度检查和含量测定,用紫外光谱(UV)、红外图谱(IR)、质谱(MS)、核磁共振氢谱(1H-NMR)、碳-13核磁共振(13C-NMR)等方法进行图谱分析及结构确证,确认分子式为C36H56O8。结果纯化制备得到的佛波双酯对照品,紫外最大吸收波长为232 nm,MS结果为高分辨电喷雾质谱(HRESI-MS)m/z 615.4120(M-H)-,HPLC检查纯度>99.5%。结论本制备方法科学有效,制备的对照品纯度符合中药化学对照品的相关要求,可作为佛波双酯原料及相关制剂质量控制的化学对照品。

Objective To study the preparation method and quality standard of the reference product of phorbol diesters(TPA),and to provide the basis for the quality control of raw materials and related preparations of phorbol diesters.Methods The methanol-water extract of croton oil was separated and purified by silica gel column preparation chromatography and reversed-phase preparation chromatography.The purity and content of the reference product of phorbol diesters were checked by high performance liquid chromatography(HPLC)and gas chromatography,and the chromatogram and structure were analyzed by ultraviolet spectroscopy(UV),infrared spectroscopy(IR),mass spectrometry(MS),Hydrogenium-1 nuclear magnetic resonance spectroscopy(1H-NMR),Carbon-13 nuclear magnetic resonance(13C-NMR)and other methods.The molecular formula was confirmed as C36H56O8.Results The maximum UV absorption wavelength of the purified reference product was 232 nm,the MS result was high-resolution electrospray ionization mass spectrometry(HESI-MS)m/z 615.4120(M-H),and the purity was>99.5%by HPLC.Conclusion The preparation method is scientific and effective,and the purity of the prepared reference product meets the relevant requirements of traditional Chinese medicine chemical reference products,and can be used as chemical reference products for the quality control of phorbol diesters raw materials and related preparations.