HPCE和HPLC测量导赤丸中盐酸小檗碱含量

Determination of Berberine from Daochi Pills by HPCE and HPLC

目的建立毛细管电泳法(HPCE)与高效液相色谱法(HPLC)测定导赤丸中盐酸小檗碱含量的方法,并比较二者在测定本品中盐酸小檗碱含量方面的差异。方法采用盐酸-甲醇(1∶100)溶液提取导赤丸样品。HPCE分离条件为:熔融石英毛细管柱(67 cm×75μm,有效长度60 cm),缓冲液体系为60 mmol.L-1磷酸二氢钾缓冲溶液-甲醇(65∶35),pH 3.5,分离电压30 kV,毛细管柱温25℃,检测波长254 nm,进样时间5 s;HPLC分离条件为:C18柱(250 mm×4.6 mm,5μm),柱温25℃,流动相为60 mmol.L-1磷酸二氢钾-甲醇(60∶40,用磷酸调pH3.5),流速1.0 mL.min-1,检测波长254 nm。结果盐酸小檗碱在HPLC和HPCE各自的分离条件下进样浓度分别在5.03～30.18μg.mL-1、30.18～80.48μg.mL-1内表现出良好线性关系(r=0.999 8;r=0.999 8),平均加样加收率分别为99.0%,100.8%,RSD分别为1.4%,0.6%。结论这两种方法均可用于导赤丸中盐酸小檗碱含量的测定,二者的测定结果稍有差异,HPCE测定盐酸小檗碱在柱效、分离时间、多组分生物碱分离上更有优势。

OBJECTIVE To develop a high performance capillary electrophoresis（HPCE） method and high performance liquid chromatography（HPLC） method for the quantitative determination of berberine in Daochi pills and evaluate two methods to find out the differences between them.METHODS Daochi pills were extracted with hydrochloric acid-methanol（1：100） solution and measured by HPCE and HPLC.An uncoated fused silica capillary（67 cm×75 μm,valid length of 60 cm） was used with a running buffer contained 60 mmol.L-1 sodium dihydrogen phosphate and methanol（65：35） under the voltage of 30 kV at 25 ℃,moderating pH 3.5 through phosphoric acid,and the detection wavelength was 254 nm.The injection time was 5 s.In HPLC method,the separation was performed on a C18 column（250 mm×4.6 mm,5 μm） under the temperature of 25 ℃ with potassium dihydrogen phosphate-methanol（60：40） as mobile phase at flow rate of 1.0 mL.min-1.The detection wavelength was set at 254 nm.RESULTS The linear calibration range of HPLC and HPCE was 5.03-30.18 μg.mL-1,30.18-80.48 μg.mL-1,relatively.Their average recoveries were 99.0% and 100.8% with RSD 1.4% and 0.6%,respectively.CONCLUSION Both methods can be applied to determine the berberine in Daochi pills,but there is a slight difference between their determination results,HPCE earned more benefits on column efficiency,shorter separation time and resolution among seperation of multicomponent.