雷公藤组培产物中雷公藤甲素和总生物碱含量的测定

建立了同一提取物中高效液相色谱法测定雷公藤甲素和紫外分光光度法测定雷公藤总生物碱含量的方法,样品用乙酸乙酯超声提取、过中性氧化铝柱,净化后用45%甲醇溶解,测定雷公藤的根皮、叶、雷公藤组培产物及培养液中雷公藤甲素和雷公藤总生物碱的含量。结果表明,雷公藤甲素的检测范围为1~100 μg/mL(R2= 0.999 9);雷公藤总生物碱在5~100 μg/mL范围内线性关系良好(R2=0.999 8);在同一提取液中,雷公藤甲素和雷公藤总生物碱的添加回收率分别为100.5%(RSD=0.63%)和99.8%(RSD=1.22%)。研究结果表明,根愈伤组织中雷公藤甲素含量为根皮粉中的1.25倍,培养液中雷公藤甲素产量占56%,总生物碱产量占65%。每生产1 g不定根培养物,雷公藤甲素产量为根皮粉中的10倍,雷公藤总生物碱产量为根皮粉中的2倍。

Study on the Determination of Triptolide and Total Alkaloids inTripterygium wilfordii Hook. f.Cultures

The content of triptolide and the total alkaloids in the root bark,leaves,the cultures and the nutrient solution were determined based on the determination of triptolide by HPLC and UV-spectroscopy method for total alkaloids in the same extract.Samples were extracted with ethyl acetate and cleaned up on an aluminum column,followed by dissolution in 45% methanol solution.The results showed that the linear relationship of the triptolide concentrations and peak areas was in the range of 1~100 μg/mL (R2=0.999 9).The linear relationship of the total alkaloids concentrations and absorbance was good in range of 5~100 μg/mL (R2=0.999 8).In the same extract the average recovery of triptolide was 100.5% (RSD=0.63%)and the average recovery of total alkaloids was 99.8% (RSD=1.22%).It's found that the content of triptolide in the root callus was about 1.25 times as that in the root bark.The yield of triptolide was 56% and that of the total alkaloids was 65% after filtering adventitious roots in the culture medium.The triptolide yield of 1 g per adventitious root was about 10 times that of root bark.The total alkaloids yield of 1 g per adventitious root was about 2 times that of root bark.