Establishment of a methodology for identifying Paeoniae Radix based on metallomic analysis.

We aimed to establish a methodology for identifying Paeonia samples based on metallomic analysis. We prepared 66 batches of samples (16 batches of crude drugs and 50 batches of cultivars, comprising 64 batches of Paeonia lactiflora and 2 batches of P. veitchii) collected from Japan and China (Inner Mongolia and elsewhere) between 1996 and 2008. P. lactiflora samples were genetically classified into white peony root (WPR) type and red peony root (RPR) type. Up to 47 elements were measured by inductively coupled plasma-mass spectrometry, and RPR type crude drug samples contained up to five times as much calcium as the others. Principal component analysis (PCA) of the multi-element fingerprints obtained suggested that P. veitchii, which grows wild, were distinguishable from the other cultivated P. lactiflora samples. This was confirmed perfectly by soft independent modeling of class analogy (SIMCA). The PCA of the fingerprints of P. lactiflora crude drug samples also suggested that it was possible to classify them by production area (Japan, Inner Mongolia, and China excluding Inner Mongolia) and genetic type (RPR and WPR types). They were also classified 100 % to the predicted class by SIMCA in both cases. These analyses were successful among the samples whose collection dates varied. This simple metallomic method is an efficient approach for verifying the complex origin of Paeoniae Radix.

Quality and safety issues related to traditional animal medicine: role of taurine.

BACKGROUND: Calculus Bovis (:C.Bovis) is one of the most precious and commonly-used medicinal materials in Japan and China. As the natural occurrence is very rare, a source of supply for C. Bovis is far behind the actual need and great efforts have been taken for some substitutes of natural C. Bovis. Unfortunately, very little information is available on the quality and/or clinical efficacy of medication based on C. Bovis. To ensure sustainable use of traditional therapeutic agents derived from C. Bovis, we felt that several issues needed to be addressed: 1) the source of the C. Bovis materials and quality control; 2) the role of taurine in the efficacy of C. Bovis. METHODS: Nine samples of natural C. Bovis and its substitutes were collected. ICP-MS was used for elemental analysis and the characterization was performed by principal component analysis (PCA) and soft independent modeling of class analogy (SIMCA) as multivariate approaches. The efficacy of C. Bovis was evaluated for morphology, viability and beating pattern on cultured cardiac myocytes and/or fibroblasts. RESULTS: PCA and multi-elemental focus was effective in discriminating C. Bovis samples derived from different habitats. A satisfactory classification using SIMCA was obtained among Australia C. Bovis, other habitats and the substitutes. Australian samples had better batch uniformity than other habitats and were composed of fewer elements. We have used Australian C. Bovis for assessment on its bioactive compounds. Rat cardiac cells incubated with C. Bovis extract (0.01-0.1 mg/ml) maintained normal morphology, viability and beating pattern. Cardiac myocytes and fibroblasts treated for 48 h with CA (0.5 mM) or DCA (0.1 mM) caused cell injury, as reflected by changes in appearance and a reduction of viability detected by the MTS assay. In cardiomyocytes, 0.5 h exposure of CA (0.5 mM) markedly decreased the velocity ratio of beating, whereas the simultaneous addition of 1 mM taurine largely prevented the decrease. CONCLUSIONS: The multi-elemental focus provided some references for the quality control and the efficacy of C. Bovis. Taurine partly attenuated the harmful actions of bile acids. It is plausible that the relationship between taurine and the bile acids contributes to therapeutic effect of C. Bovis.

The relation between clinical effects of Tokishakuyakusan and the identity of Paeonia lactiflora materials.

AIM OF THE STUDY: To investigate the relation between the clinical effects and the quality of crude drugs, we focused on Tokishakuyakusan (TS), consisted of 6 crude drugs. MATERIALS AND METHODS: We prepared two kinds of TS containing either medicinal cultivar of Paeonia lactiflora (MTS) or ornamental one (OTS). Other components were the same. First, we assessed the clinical effects of two TS formulations by cross-over study among the anemia patients. Second, we investigated the chemical differences between them by using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and Mössbauer analysis. RESULTS: The clinical effects of these formulations (3 g/day for 8 weeks) were tested in the cross-over study consisted of 12 women patients who were diagnosed as having anemia (Hb ≤ 11 g/dl) and consented to participate to this study. Both TS formulations were effective for anemia symptoms as shown by the improvement of several hematological parameters, whereas their comprehensive effects were distinguishable by Genetic Algorithm Partial Least Squares (GA-PLS) analysis. There were no significant differences in organic ingredients and Fe content measured by ultra performance liquid chromatography (UPLC) and ICP-MS, respectively. Interestingly, Mössbauer spectra of Fe ion were remarkably different between two formulations. Fe ion in MTS was only one form, but that in OTS was at least two forms. CONCLUSIONS: This study suggested that clinical effects of TS formulation reflect the quality of Paeoniae Radix.