Inhibitory Activity Against Rhizoctonia Solani and Chemical Composition of Extract from Moutan Cortex.

Rice sheath blight (RSB), caused by Rhizoctonia solani, is a significant disease of rice. The negative effects of chemical fungicides have created an urgent need for low-toxicity botanical fungicides. Our previous research revealed that the ethanol crude extract of Moutan Cortex (MC) exhibited superior antifungal activity against R. solani at 1000 µg/mL, resulting in a 100 % inhibition rate. The antifungal properties were mainly found in the petroleum ether extract. However, the active ingredients of the extract are still unclear. In this study, gas chromatography-mass spectrometry (GC-MS) was utilised for the analysis of its chemical components. The mycelium growth rate method was utilized to detect the antifungal activity. The findings indicated that paeonol constituted the primary active component, with a content of more than 96 %. Meanwhile, paeonol was the most significant antifungal active ingredient, the antifungal activity of paeonol (EC50=44.83 µg/mL) was much higher than that of ß-sitosterol and ethyl propionate against R. solani. Observation under an optical microscope revealed that paeonol resulted in abnormal mycelial morphology. This study provided theoretical support for identifying monomer antifungal compounds and developing biological fungicides for R. solani.

Ultrasound-Assisted Extraction of Paeonol from Moutan Cortex: Purification and Component Identification of Extract.

Moutan Cortex (MC) is a traditional Chinese medicine that contains abundant medicinal components, such as paeonol, paeoniflorin, etc. Paeonol is the main active component of MC. In this study, paeonol was extracted from MC through an ultrasound-assisted extraction process, which is based on single-factor experiments and response surface methodology (RSM). Subsequently, eight macroporous resins of different properties were used to purify paeonol from MC. The main components of the purified extract were identified by ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS/MS). The results indicate the optimal parameters are as follows: liquid-to-material ratio 21:1 mL/g, ethanol concentration 62%, ultrasonic time 31 min, ultrasonic temperature 36 °C, ultrasonic power 420 W. Under these extraction conditions, the actual yield of paeonol was 14.01 mg/g. Among the eight tested macroporous resins, HPD-300 macroporous resin was verified to possess the highest adsorption and desorption qualities. The content of paeonol increased from 6.93% (crude extract) to 41.40% (purified extract) after the HPD-300 macroporous resin treatment. A total of five major phenolic compounds and two principal monoterpene glycosides were characterized by comparison with reference compounds. These findings will make a contribution to the isolation and utilization of the active components from MC.

Moutan Cortex Extract Modulates Macrophage Activation via Lipopolysaccharide-Induced Calcium Signaling and ER Stress-CHOP Pathway.

Moutan Cortex, Paeonia suffruticosa root, has long been used as a medicine for the treatment of inflammatory diseases. The aim of this study was to evaluate the modulative properties of Moutan Cortex water extract (CP) on endoplasmic reticulum (ER) stress-related macrophage activation via the calcium-CHOP pathway. RAW 264.7 mouse macrophages were activated by lipopolysaccharide (LPS), and the levels of various inflammatory mediators from RAW 264.7 were evaluated. The multiplex cytokine assay was used to investigate both cytokines and growth factors, and RT-PCR was used to investigate the expressions of inflammation-related genes, such as CHOP. Data represent the levels of NO and cytosolic calcium in LPS-stimulated RAW 264.7 were significantly inhibited by CP as well as hydrogen peroxide (p < 0.05). Minutely, NO production in LPS-stimulated RAW 264.7 incubated with CP at concentrations of 25, 50, 100, and 200 µg/mL for 24 h was 97.32 ± 1.55%, 95.86 ± 2.26%, 94.64 ± 1.83%, and 92.69 ± 2.31% of the control value (LPS only), respectively (p < 0.05). Calcium release in LPS-stimulated RAW 264.7 incubated with CP at concentrations of 25, 50, 100, and 200 µg/mL for 18 h was 95.78 ± 1.64%, 95.41 ± 1.14%, 94.54 ± 2.76%, and 90.89 ± 3.34% of the control value, respectively (p < 0.05). Hydrogen peroxide production in LPS-stimulated RAW 264.7 incubated with CP at concentrations of 25, 50, 100, and 200 µg/mL for 24 h was 79.15 ± 7.16%, 63.83 ± 4.03%, 46.27 ± 4.38%, and 40.66 ± 4.03% of the control value, respectively (p < 0.05). It is interesting that the production of IL-6, TNF-α, G-CSF, MIP-1α, MIP-2, and M-CSF in LPS-stimulated RAW 264.7 were significantly inhibited by CP (p < 0.05), while the production of LIX, LIF, RANTES, and MIP-1ß showed a meaningful decrease. CP at concentrations of 25, 50, 100, and 200 µg/mL significantly reduced the transcription of Chop, Camk2α, NOS, STAT1, STAT3, Ptgs2, Jak2, c-Jun, Fas, c-Fos, TLR3, and TLR9 in LPS-stimulated RAW 264.7 (p < 0.05). CP at concentrations of 25, 50, and 100 µg/mL significantly reduced the phosphorylation of STAT3, p38 MAPK, and IκB-α in LPS-stimulated RAW 264.7 (p < 0.05). These results suggest that CP might modulate macrophage activation via LPS-induced calcium signaling and the ER stress-CHOP pathway.

Multi-elements characteristic and potential risk of heavy metals in MOUTAN CORTEX from Anhui Province, China.

To ensure the quality and safety of herbs, the content of 54 elements in MOUTAN CORTEX (MC) was determined by the ICP-AES and ICP-MS, and the health risks of Cu, As, Cd, Pb, Hg and rare earth elements (REEs) were assessed. These herbs were collected from 5 producing areas in Anhui Province, China, namely Wuhu, Tongling, Bozhou, Xuancheng and Chizhou. The multi-elements fingerprint identification of MC in Anhui Province was established. The total amount of macro-elements from Wuhu and Tongling is significantly lower than Bozhou. Among all MC from 5 producing areas, the highest content is Ca. Except for Bozhou, the content of macro-elements and REES in the other 4 origins of MC is from highest to lowest: Ca > K > Mg > Al > Fe > Na and Ce > La > Nd > Y > Pr > Er > Yb > Eu > Ho > Tb > Tm > Lu. The chemical forms of Cd in MC from Bozhou with the highest percentage were PH2O of high toxicity and migration, while the other 4 regions were PNaCl of low activity and mobility. There was a great difference in the content of inorganic elements and chemical forms of Cd between the MC produced from the plain (Bozhou) and the hilly areas (Wuhu, Tongling, Chizhou and Xuancheng). Except for Cd, the content of Cu, As, Pb and Hg in MC did not exceed the limit. The results of PTWIFact and ADI for Cd and REEs showed that MC herbs did not pose a risk to human health. Supplementary Information: The online version contains supplementary material available at 10.1007/s13762-022-04402-6.

[Chemical and nutrient differences between medicinal material, residues, and residue compost of Moutan Cortex].

Moutan Cortex(MC) residues produced after the extraction of MC can be re-extracted for active components and used to produce organic fertilizer and animal feed. However, they are currently disposed as domestic waste, which pollutes the environment. This study analyzed the chemical composition of the medicinal material, residues, and residue compost of MC by UPLC-UV-Q-TOF-MS. Furthermore, the nutrient composition of MC residues and the residue compost was analyzed. The results showed that:(1)MC residues had lower content of chemicals than the medicinal material, and content of paeonol, gallic acid, and galloylglucose in MC residues were about 1/3 of that in the medicinal material. The content of chemicals were further reduced after residue composting, and the quantitative compounds were all below the limits of detection.(2)Compared with MC residues, the residue compost showed the total nitrogen, total phosphorus, total potassium, and organic matter content increasing by 122.67%, 31.32%, 120.39%, and 32.06%, respectively. Therefore, we concluded that the MC residues can be used to re-extract active compounds such as paeonol, gallic acid, and galloylglucose. The MC residue compost is a high-quality organic fertilizer containing minimal content of chemicals and can be widely used in the cultivation of Chinese medicinal herbs.

Combining network pharmacology with chromatographic fingerprinting and multicomponent quantitative analysis for the quality evaluation of Moutan Cortex.

In this study, we combined network pharmacology, chromatographic fingerprinting and multicomponent quantitative analysis to evaluate the quality of Moutan Cortex (MC). Specifically, through network pharmacology, we obtained a comprehensive understanding of the active components and pharmacological activities of MC. In addition, the method of establishing fingerprint and multicomponent quantification by ultra high-performance liquid chromatography is convenient and comprehensive, and can more fully reflect the overall distribution of various chemical components. Principal component analysis and discriminant least square analysis were used. The results show that MC plays a synergistic role through multiple targets and pathways. The contents of chemical components in MC from different sources were significantly different. Combining network pharmacology and multicomponent quantitative results, gallic acid, benzoyl paeonol, paeonol, methyl gallate, benzoic acid and paeonol can be used as quality markers for MC quality control. This study provides a comprehensive and reliable strategy for the quality evaluation of MC and determines its quality indicators to ensure the quality of Chinese herbal medicine.

[Protective effects of Moutan Cortex polysaccharides components on renal injury in diabetic nephropathy rats].

This study investigated the protective effects of Moutan Cortex polysaccharides components(MCPC) on the renal tissues of diabetic nephropathy(DN) rats and explored their regulation effect on inflammatory response and oxidative stress. The DN rat model was induced by high-glucose and high-fat diet combined with streptozotocin(STZ), and then the rats were randomly divided into control group, model group, positive group and MCPC high(120 mg·kg~(-1)·d~(-1)), low(60 mg·kg~(-1)·d~(-1)) dose groups. After 12 weeks treatment, blood was taken from the orbit of the rats, and then they were sacrificed before the kidney tissues were collected. The serum and tissues were detected for related biochemical indicators and pathological changes of the kidney. Immunohistochemical methods were used to determine the expression of FN and ColⅣ in the kidney tissue of DN rats. Compared with the model group, blood glucose, serum creatinine, blood urea nitrogen and 24 h urine protein in the MCPC high-dose group were significantly reduced(P<0.01). The results of HE, PAS, Masson staining showed that glomerular basement membrane thickening, Bowman's capsule narrowing and inflammatory cell infiltration in DN rats were improved in the MCPC high-dose group; the activity of T-SOD and GSH-Px in serum significantly increased(P<0.001), and the expression level of FN significantly decreased(P<0.001). The high-dose MCPC treatment could effectively inhibit the abnormal expression of Col Ⅳ(P<0.001) and significantly reduce the levels of AGEs and RAGE in serum(P<0.001), the content of VCAM-1 and IL-1ß in serum(P<0.001), and the levels of IL-1ß mRNA in kidney tissue(P<0.001), but failed to effectively reduce VCAM-1 mRNA levels in kidney tissues. The high-dose MCPC could significantly improve pathological injury of renal tissue and related renal indicators in DN rats, and achieve renal protection in DN rats mainly by regulating oxidative stress and inflammatory factors.

Unveiling spatial metabolome of Paeonia suffruticosa and Paeonia lactiflora roots using MALDI MS imaging.

Paeonia suffruticosa (PS) and Paeonia lactiflora (PL) belong to the only genus in the family Paeoniaceae. Comparative analysis of the spatial metabolomes of PS and PL has rarely been performed. In this work, combined with multiple matrixes and dual-polarity detection, high mass resolution matrix-assisted laser desorption/ionization MS imaging (MALDI MSI) and MALDI tandem MSI were performed on the root sections of the two Paeonia species. The spatial distributions of many metabolites including monoterpene and paeonol glycosides, tannins, flavonoids, saccharides and lipids were systematically characterized. The ambiguous tissue distribution of the two isomers paeoniflorin and albiflorin were distinguished by tandem MSI using lithium salt doped 2,5-dihydroxybenzoate matrix. In addition, the major intermediates involved in the biosynthetic pathway of gallotannins were successfully localized and visualized in the root sections. High-mass resolution MALDI full-scan MSI provides comprehensive and accurate spatial distribution of metabolites. The analytical power of the technique was further tested in the tandem MSI of two isomers. The ion images of individual metabolites provide chemical and microscopic characteristics beyond morphological identification, and the detailed spatiochemical information could not only improve our understanding of the biosynthetic pathway of hydrolyzable tannins, but also ensure the safety and effectiveness of their medicinal use.

Comparative pharmacokinetics of seven bioactive components in normal, sham-operated, and myocardial ischemia-reperfusion injury rats after oral administration of the Salvia Miltiorrhiza-Moutan Cortex herb pair.

Recently the Salvia Miltiorrhiza-Moutan Cortex (SM-MC) herb pair is considered as a promising Chinese medicinal mixture exhibiting a range of pharmacological activities, including treating cardiovascular disease due to its unique composition. In this study, we conducted the comparative pharmacokinetic analysis of seven main bioactive components of SM-MC in a different model rat. A straightforward ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) strategy that could simultaneously evaluate the levels of seven compounds was used to ensure the reliability of these pharmacokinetic analyses in rat plasma. The rat plasma samples were collected from normal, sham-operated, and myocardial ischemia-reperfusion injury (MIRI) groups at predetermined time points after the administration of SM-MC. The main pharmacokinetic parameters were detected and calculated. We successfully assessed the maximum concentration (Cmax ), time to Cmax (Tmax ), the elimination rate constant (λz ), total half-life (t1/2 ), total body clearance (CL), and the area under the concentration-time curve from 0 to last sampling time (AUC0-t ) and extrapolated to infinity (AUC0-∞ ). To sum up, an optimized UPLC-MS/MS approach that could be used to rapidly, simultaneously, and sensitively detect seven bioactive compounds derived from SM-MC extract preparations was successfully developed, which may offer a pharmacokinetic basis for preclinical and clinical studies of SM-MC herb pair for treating MIRI.

[Protective effect of serum containing ginseng and Moutan Cortex on HUVEC injured by H_2O_2].

This study aimed to investigate the effect of serum containing ginseng and Moutan Cortex on human umbilical vein endothelial cells(HUVEC) injured with hydrogen peroxide(H_2O_2). HUVEC injured with H_2O_2 were divided into 6 groups, namely blank group, model group, ginsenoside(TGG) group, total glucosides of Moutan Cortex(TGM) group, paeonol(P) group and TGG+TGM+P group. After 24 hours of co-culture with H_2O_2, the activities of succinate dehydrogenase(SDH) and Ca~(2+)-Mg~(2+)-ATP were detected by microenzyme labeling. The apoptosis rate, intracellular Ca~(2+) concentration, reactive oxygen species(ROS) and mitochondrial membrane potential(JC-1) were detected by flow cytometry. The expressions of mitochondrial apoptosis pathway-related proteins Bax, Bcl-2, cytochrome C, caspase-3 and caspase-9 were detected by Western blot. The results showed that H_2O_2 could significantly damage HUVEC, decrease the activity of SDH and Ca~(2+)-Mg~(2+)-ATP(P<0.01), while could increase the apoptosis+necrosis rate, JC-1 decline rate, ROS increase rate and Ca~(2+) concentration increase rate(P<0.01). Serum containing ginseng and Moutan Cortex could increase the activities of SDH and Ca~(2+)-Mg~(2+)-ATP to different degrees, decrease the apoptosis+necrosis rate, JC-1 decline rate, ROS increase rate and Ca~(2+) concentration increase rate(P<0.05 or P<0.01), and down-regulate the protein expressions of Bax, caspase-3, caspase-9, cytochrome C, and up-regulate the protein expression of Bcl-2. The results showed that serum containing ginseng and Moutan Cortex has a protective effect on vascular endothelial cell injury induced by ROS, and its mechanism may be related to the improvement of mitochondrial function and the inhibition of the activation of mitochondrial apoptosis pathway.

Nine absorbed components pharmacokinetic of raw and processed Moutan Cortex in normal and blood-heat and hemorrhage syndrome model rats.

Raw Moutan Cortex (RMC) and Processed Moutan Cortex (PMC) have a long history of use in China and other Asian countries. In this study, a rapid and accurate ultra-high-pressure liquid chromatography coupled with diode array detector (UHPLC-DAD) method was developed and validated for the simultaneous determination of nine absorbed compounds of RMC/PMC. After extraction by protein precipitation with methanol from plasma, the analytes were separated on an Acquity UPLC® BEH Shield RP18 column (2.1 × 100 mm, 1.7 µm, Waters, USA). Acetonitrile (A) and 0.1% (v/v) formic acid in water (B) were selected as the mobile phase to perform gradient elution. The linearity of nine analytes was >0.9915. The intra- and inter-assay precision (RSD) values were within 11.18%, and accuracy ranged from 91.32 to 101.29%. Suitable stability, matrix effect and extraction recoveries were also obtained. The validated method was applied to compare the pharmacokinetics of RMC and PMC in Blood-Heat and Hemorrhage Syndrome Model and normal rats. The results revealed that processing and the pathological state could influence the pharmacokinetic characteristics of compounds in RMC/PMC. The study willbe useful for further studies on pharmacokinetics and clinical application of raw and processed Moutan Cortex.

[Practice and development of structural characteristics and quality control of multi-dimensional structure of basic components of genuine material of Moutan Cortex].

Traditional Chinese medicine is the product of clinical medication practice of the Chinese nation for thousands of years. Its material basis is the key to reveal the essence of the roles of traditional Chinese medicine, and the fundamental guarantee to solve the difficulties in the quality control of traditional Chinese medicine. However, the material basis of traditional Chinese medicine is to exert the overall pharmacodynamic effect through multi-targets, multi-approaches and mutual cooperation, resulting in unclear quality control index. In recent years, the quality control standards of traditional Chinese medicine have experienced great changes by shifting the focus from the appearance characteristics to the internal material basis, which however is limited to the control of a single com-ponent or multiple components. In other words, the intrinsic effectiveness and safety could not be guaranteed without the characteristics of the integrity of traditional Chinese medicine. With Moutan Cortex as an example, this paper analyzed the evolution of Moutan Cortex quality standards based on Chinese Pharmacopoeia, and comprehensively summarized the material basis of Moutan Cortex. Based on the theory of "component structure", this study analyzed current quality control of the material basis of Moutan Cortex and its preparations, and expounded the development trend of multi-dimensional quality control, so as to lay a foundation for establishing a more rational quality control system for traditional Chinese medicine in the future.

[Comparative analysis and study on component characteristics of inorganic elements in genuine Moutan Cortex and their differences from other areas].

Based on the theory of "component structure", the component structure characteristics of 17 inorganic elements in 20 batches of genuine Moutan Cortex and 12 batches of non-genuine Moutan Cortex were analyzed. The analytical method of inductively coupled plasma mass spectrometry(ICP-MS) for inorganic elements in Moutan Cortex was established, and the fingerprint of inorganic elements was drawn. Through correlation analysis and principal component analysis, the constituent elements of inorganic elements in Moutan Cortex were excavated, namely characteristic elements As, Cr, Pb, Se, K, Cu and Cd. The amount and the quantity relative ratio between the components of genuine and non-genuine Moutan Cortex samples were analyzed. It was found that the amount of characteristic elements in the bulk genuine Moutan Cortex samples was 0.33, 1.42, 1.33, 0.11, 4 385.78, 4.87, 0.49 mg·kg~(-1), with the quantity relative ratio of 1.00∶4.30∶4.03∶0.33∶13 290.24∶14.76∶1.48. The amount of characteristic elements in sub-packaged genuine Moutan Cortex was 0.24, 1.96, 0.36, 0.05, 5 122.01, 4.81, 0.05 mg·kg~(-1), with the quantity relative ratio of 1.00∶8.17∶1.50∶0.21∶21 341.71∶20.04∶0.21. The unique structural characteristics of inorganic elements in genuine Moutan Cortex are studied to provide a basis for the quality control and safety evaluation of Moutan Cortex.

[Structural characteristics of pharmacodynamical components in genuine Moutan Cortex].

Guided by the theory of "component structure", we analyzed the structural characteristics of pharmacodynamical components in genuine Moutan Cortex. The compositions of organic small molecules were determined by high performance liquid chromatography(HPLC) for 20 batches of genuine Moutan Cortex and 12 batches of non-genuine Moutan Cortex. By means of similarity analysis, clustering analysis(CA), principal component analysis(PCA) and orthogonal partial least-squares discriminant analysis(OPLS-DA), the elements in structural characteristics of the pharmacodynamical components were extracted as follows: terpene glycosides components(oxidized paeoniflorin, paeoniflorin,galloyl paeoniflorin, benzoyloxy paeoniflorinand benzoyl paeoniflorin), tannin components(1,3,6-tri-O-galloside acyl glucose, pentagalloyl glucose), and phenolic acid components(methyl gallate, paeonol). The contents and quantity ratios of terpene glycoside component, tannin component and phenolic acid components in genuine Moutan Cortex were determined as 14.1, 12.5, 21.7 mg·g~(-1), 1.00∶0.89∶1.54. The contents and quantity ratios of the oxidized paeoniflorin, paeoniflorin and benzoylpaeoniflorin in the terpene glycoside components were characteristic and determined as 2.05, 7.05, 3.30 mg·g~(-1), 1.00∶3.44∶1.61. The unique structural characteristics of genuine Moutan Cortex provide scientific basis for the formulation of quality standards.

[Quality analysis of Moutan Cortex Formula Granules based on structural characteristics of genuine components].

To evaluate the quality of Moutan Cortex Formula Granules from different manufacturers based on the structural characteristics of the genuine components of Moutan Cortex. High performance liquid chromatography(HPLC) fingerprint analysis method of Moutan Cortex Formula Granules was established to characterize the elements of the structural characteristics for genuine Moutan Cortex. Nineteen common peaks were determined and the similarity between the Moutan Cortex Formula Granules of each batch and the control fingerprint was 0.856-0.981. The results showed that there were differences in the internal quality of Moutan Cortex Formula Granules from three manufacturers. The contents of components as well as inter-component and intra-component quantity ratio for Moutan Cortex Formula Granules from different manufacturers were analyzed. It was found that the inter-component quantity ratio of G1 and G2 produced by manufacturer A was close to that of G4 produced by manufacturer B; the inter-component quantity ratio of G3 from manufacturer B was close to that of G7 from manufacturer C; and the inter-component ratios of samples G5, G6, G8-G11 produced by manufacturer C were closer to each other. The results of the study guided by the theory of "component structure" provide a new analytical method and basis for the quality evaluation of Moutan Cortex Formula Granules.

A Network Pharmacology Approach Used to Estimate the Active Ingredients of Moutan Cortex Charcoal and the Potential Targets in Hemorrhagic Diseases.

Moutan Cortex charcoal has been used to ameliorate blood heat symptoms and treat pathologic hemorrhage down the ages. Although well known as an agent with the effect of astringency and hemostasis, its active ingredients and action mechanism remain unclear. In the present study, molecular docking technology was employed to screen the potential hemostatic compounds in Moutan Cortex charcoal and their target proteins. Protein-protein-interaction (PPI) analysis was performed to explain the functions and enrichment pathways of the target proteins. The results showed that a total of 25 compounds were estimated as active constituents targeting multiple proteins related to hemostatic diseases, including 5 proteins (SERPINC1, FVIII, FX, FII and FXII) that were considered as the key targets. Then the drug-target (D-T) network was constructed to analyze the underlying hemostatic mechanism of Moutan Cortex charcoal, followed by a hierarchical cluster analysis (HCA) for compounds clustering, and a coagulation screening test for compound verification on their coagulation activities, with the results indicating that M15 (5-Tetradecenoic acid) and M31 (1-Monolinolein) might be the key compounds contributing to the hemostasis effect of Moutan Cortex charcoal by involving in the pathways related to complement, coagulation cascades and the platelet activation, particularly by activating FVIII, FX, FII and FXII and inhibiting SERPINC1. This study has demonstrated that Moutan Cortex charcoal may work as a hemostatic through the interaction between multiple-compounds and multiple-proteins, which provides the basis for further researches on the hemostasis mechanism of Moutan Cortex charcoal.

Searching for calcium antagonists for hypertension disease therapy from Moutan Cortex, using bioactivity integrated UHPLC-QTOF-MS.

INTRODUCTION: Calcium channel blockers (CCBs) are currently the most commonly used drugs for the treatment of hypertension. Moutan Cortex (MC), a traditional Chinese herb, has been found to have an anti-hypertensive effect. However, its potential mechanisms in the regulation of intracellular calcium concentration ([Ca2+ ]i ) remain poorly understood. OBJECTIVE: The main objective of this work was to identify the potential calcium antagonists from MC and study their molecular mechanisms. METHODS: Ultra-high performance liquid chromatography-quadrupole-time-of-fight-mass spectrometry (UHPLC-QTOF-MS) analysis combined with a dual-luciferase reporter assay was utilised to systematically screen the calcium antagonistic active ingredients in the methanol extract of MC. Additionally, the molecular mechanism of these compounds was further studied using live-cell imaging analysis with the calcium ion (Ca2+ ) probe dye fluo-4/AM to monitor changes in [Ca2+ ]i . RESULTS: Three monoterpenoids (paeoniflorin, benzoylpaeoniflorin and mudanpioside C), one phenolic acid (paeonol) and one gallotannin (1,2,3,4,6-O-pentagalloylglucose) were screened out as potential calcium antagonists in MC. Among them, the calcium antagonistic activity of benzoylpaeoniflorin, mudanpioside C and 1,2,3,4,6-O-pentagalloylglucose is first reported. Additionally, paeoniflorin, benzoylpaeoniflorin, mudanpioside C and paeonol can effectively block voltage-operated Ca2+ channels (VOCCs) to exert calcium antagonism, while 1,2,3,4,6-O-pentagalloylglucose plays a role in blocking inositol 1,4,5-trisphosphate receptors (IP3Rs). CONCLUSION: This work indicated that the anti-hypertensive efficacy of MC acted through multiple components selectively antagonising multiple cell signalling pathways to regulate [Ca2+ ]i . Furthermore, they could be considered as a reference standard for controlling the quality of Chinese medicinal materials.

[Application of Chinese medicine quality constant in grades evaluation of Moutan Cortex].

There are more and more literature reports about the application of Chinese medicine quality constant in the grading evaluation of Chinese medicine decoction pieces. Chinese medicine quality constant has particularly prominent advantages in comprehensive quantification,so it has become a new method and mode for grading Chinese medicine decoction pieces,highly recognized by the academic circle. In order to determine the effect of Chinese medicine quality constant,a method of grades evaluation for Moutan cortex was established in this paper. 15 batches of samples were collected from Chinese herbal slices enterprises to determine the external morphological indexes and inner quality indexes,calculate the Chinese medicine quality constant of Moutan cortex,and divide them into different grades. The results revealed that the range of the relative quality constant of these samples was from 0. 016 to 0. 060,with percentage quality constant from 27. 4 to 100. 0. If these samples were divided into three grades: the quality constant of the first grade should be ≥0. 048 or the percentage quality constant ≥80. 0; the quality constant of the second grade should be <0. 048 but ≥0. 030 or percentage quality constant <80. 0 and ≥50. 0; the quality constant of the third grade should be <0. 030 or the percentage quality constant <50. 0. This research indicated that Chinese quality constant can objectively divide grades of Moutan cortex decoction pieces,providing reference for formulating grades standards. It was also verified from the results that traditional quality evaluation of Moutan cortex was consistent with quality constant,and the connotation of percentage quality constant was elaborated as well. At the same time,it is suggested to establish a third-party Chinese medicine slices rating agency as soon as possible,which is to unify the terminology and provide rating services for the market.

[Quality evaluation of Moutan Cortex Formula Granules].

A scientific and perfect quality evaluation system for Moutan Cortex Formula Granules was established,including content determination method,characteristic chromatogram method and mass spectrometry method. The content of paeoniflorin and paeonol in Moutan Cortex Formula Granules was determined by high performance liquid chromatography( HPLC),and the average content was 1. 72% and 1. 42%,respectively. The characteristic chromatogram was used to characterize Moutan Cortex Formula Granules,which contained 7 characteristic peaks,namely gallic acid,p-hydroxybenzoic acid,oxypaeoniflorin,paeoniflorin,tetragalloyl glucose,1,2,3,4,6-penta-O-galloyl-ß-D-glucose and paeonol. A total of 40 compounds in Moutan Cortex Formula Granules,including gallic acids,paeoniflorins,paeonols,flavonoids and benzoic acids,were identified by mass spectrometry. In this study,a variety of analytical methods were used to evaluate the quality system of Moutan Cortex Formula Granules,which could play a positive role in improving the level of quality evaluation and process quality control.

[Preparation and quality standard of standard decoction of Chinese herbal medicine containing volatile components-case study of Moutan Cortex].

To prepare the standard decoction of Moutan Cortex, and research its quality standard. According to the preparation principle of standard decoction of Chinese herbal medicines, the extraction conditions were optimized, and the 14 batches of standard decoction of Moutan Cortex were prepared. The content of paeonol was determined by HPLC; the transfer rate was calculated; and the extraction rate, pH value and the fingerprint of the decoction were determined. The results showed that the transfer rate of paeonol in the standard decoction of 14 batches of samples was at the range of 43.9%-67.3%; the range of extraction rate was 22.0%-38.7%, and the pH value range was 3.98-4.77. At the same time, the fingerprints of 14 batches of Moutan Cortex standard decoction were established; 9 common peaks were determined, and the similarities were more than 0.9. The preparation method in this study was standardized with good reproducibility, and can be used for the preparation and quality standard of standard decoction of Moutan Cortex, providing reference for the quality evaluation of Moutan Cortex dispensing granules.