Abstract
A novel method of on-line 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate)-Capillary Electrophoresis-Diode Array Detector (on-line ABTS+-CE-DAD) was developed to screen the major antioxidants from complex herbal medicines. ABTS+, one of well-known oxygen free radicals was firstly integrated into the capillary. For simultaneously detecting and separating ABTS+ and chemical components of herb medicines, some conditions were optimized. The on-line ABTS+-CE-DAD method has successfully been used to screen the main antioxidants from Shuxuening injection (SI), an herbal medicines injection. Under the optimum conditions, nine ingredients of SI including clitorin, rutin, isoquercitrin, Quercetin-3-O-D-glucosyl]-(1-2)-L-rhamnoside, kaempferol-3-O-rutinoside, kaempferol-7-O-β-D-glucopyranoside, apigenin-7-O-Glucoside, quercetin-3-O-[2-O-(6-O-p-hydroxyl-E-coumaroyl)-D-glucosyl]-(1-2)-L-rhamnoside, 3-O-{2-O-[6-O-(p-hydroxyl-E-coumaroyl)-glucosyl]}-(1-2) rhamnosyl kaempfero were separated and identified as the major antioxidants. There is a linear relationship between the total amount of major antioxidants and total antioxidative activity of SI with a linear correlation coefficient of 0.9456. All the Relative standard deviations of recovery, precision and stability were below 7.5%. Based on these results, these nine ingredients could be selected as combinatorial markers to evaluate quality control of SI. It was concluded that on-line ABTS+-CE-DAD method was a simple, reliable and powerful tool to screen and quantify active ingredients for evaluating quality of herbal medicines.
Highlights
Scientists increasingly believe that reactive oxygen species (ROS) including free radicals damage the cellular constitutive elements, especially lipids and DNA in recent years1,2
ABTS+ was firstly on-line integrated with Capillary electrophoresis (CE)-DAD method to develop to quantify multiple antioxidants for evaluating the quality of herbal medicines
On-line ABTS+-CE-DAD method can be applied to evaluating the quality of TCMs preparations combining their pharmacological activity based on holistic view
Summary
Scientists increasingly believe that reactive oxygen species (ROS) including free radicals damage the cellular constitutive elements, especially lipids and DNA in recent years. LC-MS was used to measure the content of various active ingredients in previous studies, but this method cannot comprehensively evaluate the pharmacological effects and quality of SI It was reported an on-line HPLC-DAD-CL method for evaluating the quality of Ginkgo biloba leaves based on their antioxidative activity. An activity-integrated on-line 2,2-diphenyl-1-picrylhydrazyl (DPPH)-CE-DAD method was developed for comprehensively assessing antioxidative property and quality of Reduning injection in our previous research. The approach integrating DPPH with CE for measuring the total antioxidative activity of Reduning injection and the content of antioxidants in it resolved the problem of measuring activity, but the conditions of separating antioxidants and measuring the total antioxidative activity were different Taking these facts mentioned above into account, it is essential to develop a simple, rapid and reliable method for comprehensively evaluating the pharmacological effects and quality control of Shuxuening injection. In order to solve the existing problems, we developed an on-line method by integrating CE and ABTS+ for rapidly screening antioxidants from herbal medicine preparation and chose SI as the example to demonstrate the feasibility of this method
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