芯片非水毛细管电泳激光诱导荧光分离检测博落回种子中的白屈菜红碱和血根碱

目的: 建立微流控芯片非水毛细管电泳激光诱导荧光分离检测博落回种子中白屈菜红碱和血根碱的方法. 方法: 用简单玻璃十字芯片,以200 mmol·L^-1乙酸铵-乙酸-甲酰胺-水(2:0.5:5:2.5,pH=2.90)缓冲溶液作电泳缓冲液,检测距离4 cm,分离电压1.8 kV. 结果: 在150 s内即可完成白屈菜红碱和血根碱的进样和基线分离.白屈菜红碱质量浓度在0.15~550 μg·mL^-1范围内呈良好线性(r=0.9993),检测限为5.0 ng·mL^-1,平均回收率(n=6)为99.6%;血根碱质量浓度在0.3~600 μg·mL^-1范围内呈良好线性(r=0.9998),检测限为2.0 ng·mL^-1,平均回收率(n=6)为99.2%. 结论: 本方法快速、准确,样品用量少,可用于分离并定量药材中的白屈菜红碱和血根碱,并为分离结构相似的组分提供了方法借鉴.

Objective: To develop a new method for the determination of the content of chelerythrine and sanguinarine in seeds of Macleaya cordata by microchip-based nonaqueous capillary electrophoresis with laser induced fluorescence. Methods: In a simple cross glass chip,a 200 mmol·L^-1of ammonium acetate-acetic acid-formamide-water(2:0.5:5:2.5,pH=2.90) buffer solution was used as the running buffer solution,the detection length was 4 cm,and the separation voltage was 1.8 kV. Results: Baseline separation of chelerythrine and sanguinarine was achieved within only 150 s.Good linearity was observed in the range of 0.15-550 μg·mL^-1with a correlation coefficient (r) of 0.9993 for chelerythrine and in the range of 0.3-600 μg·mL^-1 with a correlation coefficient (r) of 0.9998 for sanguinarine.A low limit of detection (LOD) was achieved based on the high sensitivity of laser induced fluorescence detection (5.0 ng·mL^-1 and 2.0 ng·mL^-1 for chelerythrine and sanguinarine,respectively).The average recoveries of chelerythrine and sanguinarine were 99.6% and 99.2% respectively. Conclusion: The method showed advantages of fast analysis speed,simple process,and low consumption,which can be successfully applied to determine the contents of sanguinarine and chelerythrine in Macleaya cordata simultaneously.The method using formamide as the running buffer solution also has potential to separate other similar molecules.