Chemical Antioxidant Quality Markers of Chrysanthemum morifolium Using a Spectrum-Effect Approach

Abstract

Traditionally, the quality evaluation of Chrysanthemum morifolium (CM) cv. (Juhua) attributes its habitats and processing methods, however, this strategy of neglecting bioactive ingredients usually results in deviation of quality evaluation. This study aims to explore the quality marker (Q-marker) based on spectrum-effect relationship and quality control strategy of CMs. The chromatographic fingerprint of 30 flower head samples of CMs from five different habitats including Hang-baiju, Gongju, Huaiju, Taiju and Boju were constructed by high performance liquid chromatography and analyzed through chemometrics methods such as similarity analysis (SA), cluster analysis (CA) and principal component analysis (PCA). The common peaks were quantified by external standard method and relative correction factor method. The in-vitro radical scavenging capacity assays of DPPH·, ·OH and ABTS were carried out. The Q-marker was explored by the correlation analysis between the contents of common peaks and in-vitro radical scavenging capacity, and then used to evaluate the quality of 30 flower head samples of CMs. A total of eight common peaks were appointed in 30 flower head samples of CMs, and their similarities ranged from 0.640 to 0.956. CA results showed that 30 flower head samples of CMs could be divided into five categories with reference to the Euclidean distance of 5. PCA results showed that common peaks played a major role in differential contribution of CMs. The quantification of common peaks hinted that their contents possessed significant variation whether for different accessions or the same accessions of CMs. The correlation analysis showed that chlorogenic acid, 3,5-O-dicaffeoylquinic acid, unknown peak 1, 4,5-O-dicaffeoylquinic acid and kaempferol-3-O-rutinoside could be used as the Q-markers for the quality evaluation of 30 flower head samples of commercially available CMs. The analysis strategy that combines chromatographic fingerprint analysis, multiple ingredients quantification, in-vitro chemical anti-oxidant activity evaluation and spectrum-effect relationship analysis clarified the therapeutic material basis and discovered the Q-markers, which possibly offers a more comprehensive quality assessment of CMs.

Keywords: Chrysanthemum morifolium cv.; Q-marker; antioxidant activity; chromatographic fingerprint; spectrum-effect relationship.

FIGURE 1

Representative HPLC chromatograms of 30 flower head samples of CMs. CM represents Chrysanthemum morifolium; R means the reference average fingerprint; JH-01∼JH30 mean 30 flower head samples of different accessions of CMs and their detailed information were listed in Table 1.

FIGURE 2

Phylogenetic cluster analysis of 30 flower head samples of CMs.

FIGURE 3

Two-dimensional score plots (A) and loading plots (B) of PCA of different accessions of CMs.

FIGURE 4

Representative chromatogram of the Sample JH-07 (A) and mixed standard solution (B). 1: ChA; Peak 2, 3,5-DCQA; Peak 3, Unknown peak 1; Peak 4, L-7G; Peak 5, 4.5-DCQA; Peak 6, A-7G; Peak 7, K-3R; Peak 8, Unknown peak 2.

FIGURE 5

The box diagram of the contents of eight common peaks in 30 flower head samples of CMs.