Studies On Chemical Components Of Paeonia Veitchii And Method Of Quality Assessment Of Radix Paeoniae

Radix Paeoniae Alba (white peony root) and Radix Paeoniae Rubra (red peony root) are commonly used traditional Chinese herbal drug for analgesia, anti-inflammation and activating blood circulation to dissipate blood stasis. According to the Pharmacopoeia of the PRC (edition 2005), white peony root is the dried root of Paeonia lactiflora Pall.(peeled and boiled before drying) and red peony root is the dried root of Paeonia lactiflora Pall.or P. veitchii Lynch (directly dried). Furthermore, the dried root of Paeonia hybrida Pall is used as a substitute for red peony root in Xinjiang locally, and called as Xinjiang red peony root or red peony root.In this dissertation, the chemical components of Paeonia veitchii and method of quality assessment of Radix Paeoniae were studied systematically.From the roots of P. veitchii, 28 compounds were isolated and structurally elucidated by chemical and spectroscopic methods (¹H NMR, ¹³C NMR, HMBC, HMQC, NOESY, EI-MS, ESI-MS, HRESI-MS, IR, UV). One of them was a new compound, 2-O-[α-L-arabinopyranosyl-(1→6)-β-D-glucopyranoside]-Benzaldehyde (BH), and eighteen of them were isolated from this plant for the first time:β-sitosterol linoleate, daucosterol linoleate, linoleic acid, palmitic acid, stigmasterol,α-spinaterol, stigmast-7-en-3β-ol, friedelin, epi-friedelanol, galloylpaeoniflorin (GPF), albiflorin (AF), salicyl alcohol, isosalicin, salicin, methyl gallate (MG), pentagalloylglucose (PG), paeonol (PN), arbutin. 9 Known ones,β-sitosterol, daucosterin, paeoniflorin (PF), benzoylpaeoniflorin (BPF), oxypaeoniflorin, gallic acid (GA), benzoic acid (BA), d-catechin and saccharose, were also isolated.A reversed-phase HPLC method was developed for simultaneous determination of GA, MG, BH, AF, PF, GPF, BA, PG, BPF and PN in white peony root and red peony root, collected from different habitats and different batches. The determination were accomplished by a linear gradient elution system on ODS columns. The linear calibration curves were plotted with satisfied relative coefficients (0.9995～0.9999). The average recoveries were in the range of 96.9～104.0% with RSD less than 3.3%. The established assay method is simple, rapid, reproducible and provide a quantitative basis for the quality assessment for Radix Paeoniae.The TLC and HPLC chromatographic fingerprints of methanol extracts of white peony root and red peony root were evaluated. Totally samples derived from Paeonia lactiflora, P. veitchii and P. hybrida, respectively, were firstly analyzed by TLC documentation. The results indicated the three species of peony roots could be distinctly authenticated from each other based on their TLC characters, detected by colorimetric or fluorescent spotting, and the samples from the same species displayed higher homogenicity.The methanol extracts of the above mentioned samples were simultaneously analyzed by a newly developed HPLC system and characteristic chromatographic fmgerprints and the chemical data were acquired. The HPLC chromatographic profiles of Radix Paeoniae showing 27 diagnostic peaks from both white and red peony root and studied by chemical pattern recognition methods. The profiling data were randomly treated with hierarchical clustering analysis and divided into three classes, P. lactiflora (peeled), P. lactiflora (unpeeled), and P. veitchii plus P. hybrida. Stepwise discriminant analysis (STEPDA) method was also utilized for differentiation of Radix Paeoniae samples and four discriminant functions were produced from the result of classification, in which P. veitchii plus P. hybrida were divided into two classes in stead. When an unknown sample is analyzed by this method, the observed values are imported into the discriminant functions. The analyzed sample is identified to a specified class which give the biggest function value to the discriminant function of the specified class. Based on the results of classification, a similarity analysis method for quality assessment of Radix Paeoniae samples was developed. All the three different methods resulted in the consistent trend and can be used for identifying white peony root and red peony root.High-performance liquid chromatography with on-line photodiode array detection and electrospray ionization tandem mass spectrometry was developed for simultaneous analysis of compounds in the Radix Paeoniae. Combining the separation by HPLC, the characteristic of UV spectra, and the information of molecular structure provided by ESI-MSⁿ, eight compounds in the white peony root and red peony root were identified. Furthermore, the fragmentation pattern of them were proposed. Two inflammatory models of ear edema induced by xylene and elevation of capillary permeability by acetic acid were used for testing the anti-inflammatory effect of extracts of red and white peony roots. The effects on the activating blood and eliminating stasis with prothrombin time (PT), kaolin partial thromboplastin time (KPTT) and platelet aggregation induced by ADP were studied. The results showed that red peony root and white peony root indicated the similar biological activities in those tests.Conclusively, by using the theory and methodologies of traditional Chinese medicine (TCM), phytochemistry, analytical chemistry, chemical statistics and pharmacology, red peony root and white peony root were studied in this paper. The chemical components of Paeonia veitchii was elucidated and a new methodology for the quality control of Radix Paeoniae was established. Furthermore, the MSⁿ fragmentation patterns of compounds were proposed and a comparative study on pharmacological effects was carried out, which provided valuable academic evidences for the future research and development of Radix Paeoniae.