Wang Zhiying, Guo Haitao, Chen Meng, Zhang Guangbin, Chang Ruimiao, Chen Anjia
College of Pharmacy, Shanxi Medical University, Taiyuan, P. R. China.
Electrophoresis. 2018 Sep;39(17):2195-2201. doi: 10.1002/elps.201800129. Epub 2018 Jul 8.
A sensitive, fast, and effective method, field-amplified sample stacking (FASS) in capillary electrophoresis, has been established for the separation and determination of corynoxine and corynoxine B. Hydroxypropyl-β-CD (HP-β-CD) and tetrabutylammonium-L-glutamic acid (TBA-L-Glu) were used as additives in the separation system. Electrokinetic injection was chosen to introduce sample from inlet at 10 kV for 50 s after a water plug (0.5 psi, 4 s) was injected to permit FASS. The running buffer (pH 6.1) was composed of 40 mM sodium dihydrogen phosphate solution, 130 mM HP-β-CD, and 10 mM TBA-L-Glu and the separation voltage was 20 kV. Under the optimum conditions, corynoxine and corynoxine B were successfully enriched and separated within 12 min and the sensitivity was improved approximately by 700-900 folds. Calibration curves were in a good linear relationship within the range of 62.5-5.00 × 10 ng/mL for both corynoxine and corynoxine B. The limits of detection (S/N = 3) and quantitation (S/N = 10) were 14.9, 45.2 ng/mL for corynoxine and 11.2, 34.5 ng/mL for corynoxine B, respectively. Finally, this method was successfully applied for the determination of corynoxine and corynoxine B in the stems with hooks of Uncaria rhynchophylla and its formulations.
已建立一种灵敏、快速且有效的方法——毛细管电泳中的场放大进样堆积(FASS),用于分离和测定钩藤碱和异钩藤碱。羟丙基-β-环糊精(HP-β-CD)和四丁基铵-L-谷氨酸(TBA-L-Glu)用作分离体系中的添加剂。选择电动进样,在注入水塞(0.5 psi,4 s)以实现FASS后,于10 kV下从进样口引入样品50 s。运行缓冲液(pH 6.1)由40 mM磷酸二氢钠溶液、130 mM HP-β-CD和10 mM TBA-L-Glu组成,分离电压为20 kV。在最佳条件下,钩藤碱和异钩藤碱在12分钟内成功富集并分离,灵敏度提高了约700 - 900倍。钩藤碱和异钩藤碱在62.5 - 5.00×10 ng/mL范围内校准曲线呈良好线性关系。钩藤碱的检测限(S/N = 3)和定量限(S/N = 10)分别为14.9、45.2 ng/mL,异钩藤碱的检测限和定量限分别为11.2、34.5 ng/mL。最后,该方法成功应用于钩藤及其制剂带钩茎中钩藤碱和异钩藤碱的测定。