Abstract

This paper describes the development of an HPLC-UV-MS method for quantitative determination of andrographolide and dehydroandrographolide in Andrographis Herba and establishment of its chromatographic fingerprint. The method was validated for linearity, limit of detection and quantification, inter- and intra-day precisions, repeatability, stability and recovery. All the validation results of quantitative determination and fingerprinting methods were satisfactory. The developed method was then applied to assay the contents of andrographolide and dehydroandrographolide and to acquire the fingerprints of all the collected Andrographis Herba samples. Furthermore, similarity analysis and principal component analysis were used to reveal the similarities and differences between the samples on the basis of the characteristic peaks. More importantly, the DPPH free radical-scavenging and ferric reducing capacities of the Andrographis Herba samples were assayed. By bivariate correlation analysis, we found that six compounds are positively correlated to DPPH free radical scavenging and ferric reducing capacities, and four compounds are negatively correlated to DPPH free radical scavenging and ferric reducing capacities.

Highlights

  • Andrographis Herba (AH), the aerial part of Andrographis paniculata

  • The mean recovery (n = 5) are 107.02% ± 1.52% and 98.86% ± 1.46%, respectively. These RSD values indicate that the proposed methodology is reproducible and suitable for the quantitative determination of andrographolide and dehydroandrographolide in AH samples

  • The developed HPLC-UV analytical method was applied for the quantitative determination of andrographolide and dehydroandrographolide in ten AH samples

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Summary

Introduction

Andrographis Herba (AH), the aerial part of Andrographis paniculata (Burm. F.) Nees, has been widely used for clearing heat and removing toxicity, cooling blood and detumescence [1]. Due to its efficacy and affordability, the raw herb material is popular and is widely used in many Chinese herbal compound prescriptions. Andrographolide and dehydroandrographolide are used as chemical markers for quality control of AH in the Chinese Pharmacopoeia (2010 edition) which stipulates that the total content of the two markers should not be less than 0.8% in qualified AH samples. The correlation between chemical compounds and therapeutic effects of an herb is usually not elaborated. Based on this idea, a chromatographic fingerprinting method using HPLC-UV-MS was developed in the present study. Antioxidant capacities of AH samples were tested and fingerprint-efficacy correlation was studied for the first time

Optimization of the Extraction Method
Optimization of Chromatographic Conditions
Validation of Quantitative Analytical Method
Validation of the Chromatographic Fingerprinting Method
Quantitative Determination of the Two Analytes in AH Samples
Assignments of the Characteristic Peaks
Fingerprinting and Chemometrics Analyses
DPPH Assay
FRAP Assay
Correlation Analysis
Experimental Section
Sample and Reference Preparation
HPLC Analysis
HPLC-ESI-MS Analysis
Similarity Analysis
Conclusions
Conflicts of Interest
Full Text
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