The Activity-Integrated Method for Quality Assessment of Reduning Injection by On-Line DPPH-CE-DAD

Abstract

A sensitive on-line DPPH-CE-DAD method was developed and validated for both screening and determining the concentration of seven antioxidants of Reduning injection. The pH and concentrations of buffer solution, SDS, β-CD and organic modifier were studied for the detection of DPPH and seven antioxidants. By on-line mixing DPPH and sample solution, a DPPH-CE method for testing the antioxidant activity of the complex matrix was successfully established and used to screen the antioxidant components of Reduning injection. Then, antioxidant components including caffeic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, chlorogenic acid, neochlorogenic acid and cryptochlorogenic acid were quantified by the newly established CE–DAD method. Finally, the total antioxidant activity and the multiple active components were selected as markers to evaluate the quality of Reduning injection. The results demonstrated that the on-line DPPH-CE-DAD method was reagent-saving, rapid and feasible for on-line simultaneous determination of total pharmacological activity and contents of multi-components samples. It was also a powerful method for evaluating the quality control and mechanism of action of TCM injection.

A Green Antioxidant Activity-Integrated Dual-Standard Method for Rapid Evaluation of the Quality of Traditional Chinese Medicine Xuebijing Injection by On-Line DPPH-CE-DAD

“…The running electrolyte for sample analysis was similar to that described previously [18], except that it was made possible to simultaneously separate and quantify DPPH and the antioxidants in one step by the newly DPPH-CE-DAD method.…”

“…Based on our previous research [18], 20 mM Na 2 HPO 4 (pH 6.0) + 50 mM SDS was the optimized condition for analyzing DPPH. However, an optimal condition comprising an electrolyte containing 20 mM NaH 2 PO 4 (pH 5.5), 100 mM SDS, 10 mM β -CD, and 5% ACN (v/v) with the voltage and temperature setting at 25 kV and 22°C was employed in all of our subsequent experiments, thereby reaching a balance between evaluating total antioxidant activity and screening antioxidants of Xuebijing injection in one step.…”

“…These pharmacological methods can lead to long cycle, false positives and can be harmful to the environment. Recently, high-performance capillary electrophoresis (HPCE), as a green separation technology, offers an alternative method that has many advantages compared to other methods, in that it provides good resolution of the sample with a shorter run time, uses less harmful solvents, and is not influenced by color pigments [17, 18]. …”

“…In a previous paper, we reported our preliminary finding that the on-line DPPH-CE-DAD conditions for determination of the total antioxidant activity and contents of antioxidants were entirely different [18]. If two conditions can be integrated into the same condition, thus the total time needed for the experiment will be significantly cut down and less intensive labor will be achieved.…”

The in‐capillary DPPH‐capillary electrophoresis‐the diode array detector combined with reversed‐electrode polarity stacking mode for screening and quantifying major antioxidants in Cuscuta chinensis Lam

“…The oxidation reaction was carried out by off‐line incubation, which was time consuming and sample consuming. As an alternative, in‐capillary DPPH‐CE was proposed to screen antioxidants from mixtures, in which the reactants (DPPH and sample solution) were successively injected into the capillary, then mixed by electrophoresis and incubated to generate sufficient product in our previous work . It was demonstrated that an in‐capillary DPPH‐CE‐DAD method could simultaneously determine total antioxidants of samples and screen active components rapidly from the complex in two separation conditions.…”