阔叶十大功劳总生物碱提取及小檗碱含量测定

Study of Extraction for Total Alkaloids and the Content Determination of Berberine in Mahonia bealei

目的建立阔叶十大功劳中总生物碱的提取工艺和小檗碱的含量测定方法。方法通过正交试验确立提取总生物碱的最佳工艺条件,紫外分光光度法测定总生物碱含量;采用HPLC-UVD法,C18色谱柱（4.6 mm×250 mm,5μm）,以甲醇-水（各含0.1%甲酸）为流动相,流速1 m L/min,柱温35℃,检测波长345 nm,测定小檗碱的含量。结果最佳提取工艺为药材质量6倍量的80%乙醇、提取4次、每次2 h;小檗碱在36.00~486.00μg/m L范围内呈良好线性关系,平均回收率为98.0%,RSD为0.53%。结论总生物碱的提取工艺回收率较高、稳定且重复性好;所建立的测定小檗碱含量的HPLC法简便、快速,结果准确可靠。

Objective To establish the extraction technology of total alkaloids and the method for determination of berberine in Mahonia bealei. Methods The optimum extraction technology of total alkaloids from Mahonia bealei was determined by orthogonal experiment. The content of total alkaloids was determined by ultraviolet spectrophotometry; The content of berberine was determined by HPLC-UVD method. The method was developed on a C18column（4.6 mm×250 mm,5 μm） by gradient elution with methanol-water（both containing 0.1‰（V/V） formic acid） as mobile phase at a flow rate of1 m L·min-1with the column temperature at 35 ℃ and the detection wavelength was at 345 nm. Results The optimal extraction technique was to extract pueraria in 80% alcohol with 6 times the weight of herbal medicine for 4 times, with2 hours once; There was good linearity in the range of 36~486 μg·m L-1for berberine; The average recovery of berberine was98.03% and RSD was 0.53%（n=6）. Conclusion High content of total alkaloids are obtained with the present technology. The results with better repeatability are stable; The methods for determination of the content of berberine by HPLC was proved to be simple and the results obtained were reliable and suitable. The study can provide the reference for extraction of total alkaloids in M. bealei and determination of the content of berberine.