论文部分内容阅读
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 establish a method for the determination of berberine hydrochloride in Zhishu Pills by capillary electrophoresis (HPCE) and high performance liquid chromatography (HPLC), and to compare the differences between the two in determining the content of berberine hydrochloride in this product. Methods Chizhwan Pill was extracted with hydrochloric acid - methanol (1: 100) solution. HPCE separation conditions: fused silica capillary column (67 cm × 75 μm, effective length 60 cm), buffer system 60 mmol.L-1 potassium dihydrogen phosphate buffer solution-methanol (65:35), pH 3.5, separation voltage The column temperature was 25 ℃ and the mobile phase was 60 mmol·L-1. The chromatographic conditions were as follows: C18 column (250 mm × 4.6 mm, 5 μm) 1 potassium dihydrogen phosphate - methanol (60:40, pH3.5 with phosphoric acid), the flow rate of 1.0 mL.min-1, detection wavelength 254 nm. Results The berberine hydrochloride showed a good linearity in the range of 5.03 ~ 30.18μg.mL-1 and 30.18 ~ 80.48μg.mL-1 under the conditions of HPLC and HPCE respectively (r = 0.999 8; r = 0.999 8). The average addition rates were 99.0% and 100.8%, respectively, with RSDs of 1.4% and 0.6%, respectively. Conclusions Both of these methods can be used to determine the content of berberine hydrochloride in Zhidiao Pill. The results of the two methods are slightly different. The determination of berberine hydrochloride by HPCE is more effective in column efficiency, separation time and multi-component alkaloid separation Advantage.