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目的:建立UV、HPLC测定广东钩藤属4种植物的总碱以及钩藤碱、异钩藤碱含量的方法,比较其不同品种、不同部位间有效成分的含量变化。方法:以UV测定总碱含量,检测波长为242 nm;以HPLC测定钩藤碱、异钩藤碱含量,采用Hypersil ODS C18(4.6 mm×200 mm,5μm)色谱柱,甲醇-水(含0.50%三乙胺,冰乙酸调pH 6.33)=63∶37为流动相,流速为0.80 mL/min,进样量5.00μL,柱温为室温,245 nm波长下检测分析。结果:钩藤碱、异钩藤碱在0.02～5.00μg范围内线性关系良好,钩藤碱的r1=0.9999,异钩藤碱的r2=1.0000,平均回收率分别为98.52%、99.12%(RSD1=2.05%,RSD2=2.17%);总碱含量在不同品种、不同部位里存在差异性,其线性范围为6.00～14.00μg/mL,r=0.9989。结论:该法准确,操作简便、快速,重复性较好,精密度较高,可用于对广东钩藤属植物不同品种、不同部位的总碱以及钩藤碱、异钩藤碱含量测定。
OBJECTIVE: To establish a method for the determination of total alkaloids, rhynchophylline and isorhamnetin in four species of Uncaria in Guangdong by UV and HPLC, and to compare the contents of active constituents in different varieties and parts. Methods: The total alkaloid content was determined by UV at a wavelength of 242 nm. The contents of rhynchophylline and isorhamine were determined by HPLC. The content of rhynchophylline and isorhamnetin were determined by HPLC on a Hypersil ODS C18 column (4.6 mm × 200 mm, 5 μm) % Triethylamine, glacial acetic acid to adjust the pH to 6.33) = 63:37 as mobile phase, the flow rate was 0.80 mL / min, the injection volume was 5.00 μL, the column temperature was room temperature, 245 nm wavelength detection analysis. Results: The linear relationship between rhynchophylline and isorhynchophyll in the range of 0.02 ~ 5.00μg was good, with the r1 = 0.9999 for rhynchophylline and r2 = 1.0000 for isorhamnetine. The average recoveries were 98.52% and 99.12% (RSD1 = 2.05%, RSD2 = 2.17%). The total alkaloid content varied in different varieties and different parts with the linear range of 6.00-14.00μg / mL, r = 0.9989. Conclusion: The method is accurate, simple, rapid, reproducible and accurate. It can be used to determine the content of total alkaloids, rhynchophylline and isorhamnetin in different varieties and parts of Uncaria.