Exploring potential quality markers of Paeoniae Radix Alba by UPLC-Q-TOF/MS, HPLC, and cell experimentation

Abstract

Paeoniae Radix Alba (PRA), the dried root of Paeonia lactiflora Pall., has been used in traditional Chinese medicine for thousands of years. Nevertheless, the potential quality markers (Q-markers) of PRA remain unclear and establishing a reliable quality evaluation method for PRA based on the serum pharmacochemistry theory is challenging. In this study, ultra-high-performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to identify the chemical components in PRA extract and in rat serum after oral administration of PRA. High-performance liquid chromatography (HPLC) was used to develop the chromatographic fingerprint and quantitative analysis of PRA. In addition, a lipopolysaccharide (LPS)-induced RAW 264.7 cell inflammatory model was used to determine the anti-inflammatory effects of the potential Q-markers of PRA. A total of 53 chemical components were identified in PRA extract, and 14 prototype ingredients and 3 metabolites were absorbed into the blood, which may be the potential active components of PRA. Thirteen common peaks were detected in the fingerprints of the 12 batches of PRA samples from different origins, 5 of which were blood-entering ingredients and were quantitated. Cell experiments confirmed that these five compounds, including oxypaeoniflorin, albiflorin, paeoniflorin, lactiflorin, and palbinone, could effectively alleviated the generation of NO, IL-1β, IL-6, and TNF-α in LPS-induced RAW 264.7 cells. These results indicated that oxypaeoniflorin, albiflorin, paeoniflorin, lactiflorin, and palbinone may serve as the potential Q-markers for PRA, which could be helpful for the quality control of the production, processing, and preparation of decoction pieces and Chinese patent medicine of PRA.