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目的建立测定明党参原植物及离体培养材料中3个代表性呋喃香豆素成分花椒毒素、佛手酚和佛手柑内酯的高效液相色谱方法。方法采用HPLC-PDA,Agilent-C18色谱柱(4.6 mm×250 mm,5μm),流动相为甲醇-水梯度洗脱,流速1.0 m L·min~(-1),检测波长314 nm,进样量10μL。结果所建方法可同时测定明党参样品中3种香豆素含量,花椒毒素、佛手酚和佛手柑内酯的线性方程分别为Y=17 057ρ-87.689(r=1.000 0)、Y=23 828ρ-380.44(r=0.999 9)、Y=37 123ρ-441.16(r=0.999 9)。原植物和再生植株中花椒毒素与佛手柑内酯含量明显高于愈伤组织和悬浮细胞,但佛手酚在前两者中未检出而在后两者中有少量积累,源于叶和柄细胞的样品3种香豆素含量普遍较高。结论所建方法用于测定明党参中3种香豆素含量,准确、灵敏、重复性良好。该法为明党参资源有效开发与利用,尤其为利用生物技术进一步探索香豆素类有效成分的调控奠定基础。
OBJECTIVE To establish a HPLC method for the determination of 3 representative furanocoumarin components, pricklyashin, bergapten and bergamantone, from the original plants of Codonopsis pilosula and cultured in vitro. Methods The HPLC-PDA and Agilent-C18 column (4.6 mm × 250 mm, 5 μm) were used. The mobile phase consisted of a gradient of methanol and water with a flow rate of 1.0 m L · min -1. The detection wavelength was set at 314 nm. Amount of 10 μL. Results The established method could be used to determine the contents of three coumarins in the samples of Codonopsis pilosula. The linear equations of xanthotoxin, bergamol and bergamot were Y = 17 057ρ-87.689 (r = 1.000 0), Y = 23 828ρ -380.44 (r = 0.999 9), Y = 37 123ρ-441.16 (r = 0.999 9). The content of pricklyashin and bergamot lactone in the original plants and the regenerated plants was significantly higher than that in the callus and the suspension cells, but the bergamol was not detected in the first two and a small amount accumulated in the latter two, The samples of the cells showed generally high levels of 3 coumarins. Conclusion The proposed method was used to determine the content of three coumarins in Codonopsis Pilosula, which is accurate, sensitive and reproducible. The method for the effective development and utilization of Codonopsis resources, especially for the use of biotechnology to further explore the regulation of the active ingredients of coumarin laid the foundation.