A novel method of reverse migration micellar electrokinetic chromatography (RM-MEKC) using 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulphonate) as oxidation-free radical was developed to screen the major antioxidants from Sanyetangzhiqing (SYTZQ), which is a new patent drug for diabetes. For simultaneous detection and separation of 2,2′-bis (3-ethylbenzothiazoline-6-sulfonic acid) ammonium salt (ABTS + ) and chemical components of SYTZQ, the key factors were optimized and the method was validated under the optimal conditions. All the relative standard deviations of recovery, precision, and stability were below 6.6%. Based on these, the RM-MEKC method has been successfully used to screen the main antioxidants from SYTZQ tablets. Five compounds, including rosmarinic acid, salvianolic acid B, lithospermic acid, hyperoside, and isoquercetin, were separated and identified as the major antioxidants. There was a linear relationship with correlation coefficient of 0.923 between the total amount of major antioxidants and total antioxidative activity of SYTZQ. Consequently, these five ingredients could be selected as combinatorial markers for quality control of SYTZQ, and the established method was concluded to be a simple, reliable, and powerful tool to screen and quantify active compounds for quality evaluation of SYTZQ.

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Award Number: 21ZYJDJC00080An in-capillary 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulphonate) as oxyradicals combining field-enhanced sample injection with micellar electrokinetic chromatography was developed for screening and determination of the major antioxidants in Yangxinshi Tablet. To obtain simultaneous separation and detection of radicals and analytes, relevant factors were optimized separately. Under the optimum conditions, four compounds including salvianolic acid B, hyperoside, puerarin, and caffeic acid were identified as the major antioxidants. All validation results covering recovery, precision, and stability demonstrated good applicability of the method. On this basis, the total antioxidant activity was successfully evaluated in terms of the decreased peak area of radicals. There was a correlation coefficient of 0.8974 between the total contents of major antioxidants and the total antioxidative activity of the sample. Therefore, these four compounds were selected as combinatorial markers for the quality evaluation of Yangxinshi Tablet. It was concluded that the established method presented a powerful potential to screen and quantify active ingredients in the complex preparation of Chinese medicine.

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A simple and sensitive strategy using cyclodextrin-modified micellar electrokinetic chromatography with diode array detector was developed and applied for the simultaneous separation and determination of nine components in Sanyetangzhiqing (SYTZQ), a hypoglycemic and hypolipidemic agent. Several important parameters affecting separation performance were evaluated and optimized using single variable methods. Under the optimal conditions, baseline separation of the nine components, including four flavonoids (hyperoside, isoquercitrin, quercetin-3-O-glucuronoside, and astragalin), four phenolic acids (chlorogenic acid, rosmarinic acid, salvianolic acid B, and lithospermic acid), and a monoterpenoids (paeoniflorin), were achieved in less than 16 min. The correlation coefficients of the calibration curves were over 0.9996 for all the analytes. Intraday and interday precisions ranged from 0.4% to 4.8% and 1.7% to 5.0%, respectively. Recoveries of analytes varied from 95.3% to 105%. Validation results as well as the application to analyse SYTZQ samples demonstrated the applicability of the proposed method and thus provided an effective tool for the quality control of SYTZQ. Moreover, with the advantages of short time consuming, low energy consumption, high efficiency, and low cost, this method has laid a foundation for the determination and quality evaluation of multicomponents in Chinese herbal compounds.