Screening and Identification of Antioxidative Material Basis from Prinsepia utilis Royle Based on the Spectrum-effect Relationship

To investigate the antioxidant chemical constituents of Prinsepia utilis Royle and reveal its pharmacodynamic substance basis. HPLC was used to establish the fingerprints of different batches of Prinsepia utilis Royle extracts (PRE). The DPPH scavenging activity was used as an indicator of drug efficacy. Pearson correlation, Spearman analysis, and partial least squares regression (PLSR) analysis were used to study the spectral efficiency relationship between the common peaks of the HPLC fingerprint spectra and the antioxidant efficacy substances of PRE. This was followed by LC-MS and HPLC analyses to identify the primary antioxidant substances. We assessed the antioxidant and anti-aging effectiveness of these active substances through various methods: scavenging 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH·), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid radicals (ABTS⁺·), OH radicals (·OH), and anti-aging activity in HFF cells. Results showed that thirteen common peaks were indexed and six chemical components were identified from the fingerprints of 11 batches of PRE. Among them, rutin, narcissoside, clitorin, shaftoside and typhaneoside may be potential antioxidant substances in the 11 extracts of PRE, and the contents of the above five compounds exhibited significant differences. In which, the content of clitorin was the highest (738.123 μg/mL), followed by rutin (538.612 μg/mL), narcissoside (385.881 μg/mL), and typhaneoside (337.168 μg/mL). Rutin and narcissoside demonstrates significant efficacy in scavenging DPPH·, ABTS⁺·, and ·OH radicals, with the scavenging rates and IC₅₀ values recorded at DPPH· 92.84% (Rutin) and 37.71% (narcissoside), ABTS⁺· 92.44% (Rutin) and 82.01% (narcissoside) , ·OH 0.45 (Rutin) and 1.01 (narcissoside) mg/mL, respectively. Additionally, schaftoside, typhaneoside, and rutin significantly inhibited SA-β-gal activity, reduced the proportion of senescence-associated β-galactosidase-positive cells, and exhibits notable anti-aging effects (P<0.01). In the hydrogen peroxide (H₂O₂)-induced cellular oxidative stress model, typhaneoside, clitorin, narcissoside, and shaftoside, along with rutin significantly downregulated the expression of MMP-1, MMP-3, and MMP-9 (P<0.001), promoted skin collagen synthesis, and inhibited its degradation, demonstrating potential anti-aging capabilities. This altogether indicates that the antioxidant effect of Prinsepia utilis Royle is the result of the combined effect of multiple chemical components such as shaftoside, typhaneoside, rutin, narcissoside and clitorin.