北豆根提取物中有效成分分析方法的建立

Establishment of Analytic Method of Extracting Effective Ingredients from Rhizoma Menispermi(RM)

目的采用酸性染料比色法测定北豆根提取物中北豆根总碱（TAMD）的含量，采用高效液相色谱（HPLC）法测定蝙蝠葛碱（Dauricine）的含量。方法溴甲酚绿在无水甲醇（pH=6）中，可与TAMD直接络合生成绿色化合物，用于比色测定，最大吸收波长为620nm。HPLC法，色谱柱：Discovery C18柱；流动相：乙腈：水：磷酸=16：84：0．15；流速：0．5ml·min^-1；检测波长：208nm。结果酸性染料比色法：线性范围为6～40μg·ml^-1，r=0．9998（n=7），平均加样回收率为99．18％，RSD=0．74％；HPLC法：线性范围为0．01～0．1mg·ml^-1，r=0．9998（n=7），平均加样回收率为98．32％，RSD=0．94％。结论建立的方法灵敏、准确、重现性好，可作为北豆根提取物中TAMD和蝙蝠葛碱的含量分析方法。

Objective To establish an acid dye eolorimetric method for determination of total alkaloid in Menispermum dauricum （TAMD）, an HPLC method for determination of Dauricine . Methods TAMD produced green combination with bromocresol green in the condition of dehydrated ethanol , according to this characteristic, acid dye colorimetry was established. HPLC method：The separation was carried out on a Discovery -C18 column（250 mm ×4.6 mm,5 μm） , the mobile phase was acetonitrile -water - phosphoric acid（ 16： 84：0.15 ） , the flow rate was 0.5 ml · min^ -1 , and detection wavelength was 208 nm. Results Acid dye colorimetric method：The linear range was 6 -40 μg · ml^-1 , r =0.999 8（n =7） , the average recovery was 99.18% with the RSD of 0.74%. HPLC method：The linear range was 0.01 - 0.1 mg · ml^-1 , r = 0. 999 8 （ n = 7） ,the average recovery was 98.32% with the RSD of 0.94%. Conclusion This method is sensitive, accurate and reproducible, and can be used in analyzing the content of TAMD and Dauricine in RM extract.