Background : Spiraea prunifolia var. simpliciflora (Rosaceae) called “Brial wreath” is a deciduous latifoliate shrub that is widely distributed throughout in Northeast Asia. Phytochemical and biological investigation of S. prunifolia have led to the discovery of biologically active compounds. Pharmacological studies revealed that the extract of the root of S. prunifolia possess antioxidant, antipyretic and anti-inflammatory activities. Some chemical constituents such as sterols, phenolics, terpene and fatty acid, as well as ethanolic extracts from the roots of S. prunifolia, have previously been reported to modulate the deleterious effects of diabetes, to prevent high-fat diet-induced obesity, and to prevent cisplatin-induced nephrotoxicity. Our continuing research was carried out to search for other phytochemical constituents from the leavess of S. prunifolia. The chemical structures of compounds were determined by NMR and FAB/MS spectroscopic data interpretation. Methods and Results : Multiple-preparative liquid chromatography (MPLC) purifications were carried out on YMC LC-forte/R instrument (YMC Kyoto, Japan) equipped with YMC-Pack ODS-A columns (ODS gel : 5 ㎛, 10 ㎜ × 250 ㎜). High-performance liquid chromatography (HPLC) was performed on Agilent Technologies instrument (Aglient Tec., Santa clara, CA, USA) equipped with YMC–Pack Pro C18 columns (ODS gel : 5 ㎛, 4.6 ㎜ × 250 ㎜). Next, quantitative analysis was carried out on UPLC-QqQ/MS 3200 Q-TRAP instrument (AB SCIEX Toronto, Canada) using a ACQUITY UPLC (waters corp.) with an ACQUITY BEH C18 column (2.1 ㎜ × 100 ㎜, 1.7 ㎛). The metabolite samples was applied to preparative reversed-phase HPLC and UPLC using gradient method, solvent A [water + 0.1% formic acid (v/v)] and solvent B [acetonitrile + 0.1% formic acid (v/v)]. Conclusion : In this study, we isolated the major metabolites from the stem of Spiraea prunifolia var. simpliciflora by using MPLC and HPLC. UPLC-QqQ/MS was also used to quantify target compounds. Finally, we established methodology and performed the quantitative analysis on target compounds from the stem of Spiraea prunifolia var. simpliciflora.

Background : The development of an antioxidant to prevent disease by ROS-induced oxidative stress is necessary. This study investigated the changes of antioxidant capacities of two medicinal crops extracts by lactic acid fermentation. Methods and Results : The changes of free-radical scavenging activity of medicinal crops extracts by lactic acid fermentation were evaluated by using DPPH free-radical scavenging assay and ABTS free-radical scavenging assay. The DPPH free-radical scavenging activity of extracts or lactic acid fermented extracts were estimated as followed. samples were thoroughly mixed with 1 ㎖ ethanol solution of 0.1 mM DPPH. After stand for 30 min in the dark, the absorbance was measured at 570 ㎚ by using a UV Spectrophotometer. ABTS scavenging activity of extracts or lactic acid fermented extracts were estimated as followed. The working solution was prepared by mixing 1 ㎖ of ABTS solution with 88 ㎖ of 50% ethanol. A total of 25 ㎕ of samples were mixed with 225 ㎕ of ABTS working solution and allowed to stand for 10 min. The absorbance was read at 732 ㎚ in a UV spectrophotometer. The data were showed that lactic acid fermented extracts were higher antioxidant ability than the extracts. Conclusion : This study was showed that the antioxidant capacities of two medicinal crops extracts were improved by lactic acid fermentation.

Background : While the anti-inflammatory effects of 20 (S)-ginsenoside Rg3 (Rg3) have been studied, it remains unclear how Rg3 regulates lipid metabolism in inflammatory macrophages. Thus, in this study, we characterized some eicosanoids related to the anti-inflammatory effects of 20 (S)-ginsenoside Rg3 in murine macrophages. Methods and Results : UPLC-MS/MS was used to profile various eicosanoids from RAW264.7 cells treated with lipopolysaccharide (LPS) and Rg3. The profiling data were statistically analyzed by principal component analysis, hierarchical clustering analysis and analysis of variance. The anti-inflammatory effect of Rg3 was validated by assessing the levels of nitric oxide, tumor necrosis factor-α, and interleukin-6 in the activated macrophages treated with Rg3. A total of 69 eicosanoids were analyzed in RAW264.7 cells. Principal component and hierarchical cluster analyses differentiated control cells from cells treated with LPS, Rg3, or LPS + Rg3 for 12 or 24 h. Furthermore, some differentially regulated compounds were found between macrophages treated with LPS for 24 h and those treated with LPS + Rg3 for 24 h. Conclusion : Rg3 alters eicosanoid metabolism in activated macrophages treated with LPS. Furthermore, we identified several eicosanoids correlated with the anti-inflammatory activity of Rg3.

Background : Methicillin-Resistant Staphylococcus aureus (MRSA) is a multidrug-resistant (MDR) strain. Especially, MRSA is developing resistance to available antibacterial agents and causing complications in the treatment of infections related to skin, soft tissue, respiratory, bone, joint, and endovascular disorders. Therefore, antibacterial agent combination therapy appears to be a useful option, particularly in developing countries where antibiotic availability is limited. (+)-Usnic acid (UA) is uniquely found in lichens, and is especially abundant in genera such as Usnea and Cladonia. UA has antimicrobial activity against human and plant pathogens. Therefore, UA may be a good antibacterial drug candidate for clinical development. Methods and Results : In search of a natural products capable of inhibiting this multidrug-resistant bacteria, we have investigated the antimicrobial activity of UA against MRSA. In this study, the effects of a combination of UA and permeable agents against MRSA were investigated. For the measurement of cell wall permeability, UA with concentration of Ethylenediaminetetraacetic acid (EDTA) was used. In the other hand, Sodium azide (NaN3) was used as inhibitors of ATPase. These results suggest that the antibacterial effect of UA was potentiated by membrane-binding agents and ABC transporter-inhibiting agents, implying that antibacterial activity is associated with damage of the cell wall and inhibition of ATPase function by UA. Conclusion : UA and in combination with EDTA and NaN3 could lead to the development of new combination antibiotics against MRSA infection. The results of this study appear to be promising, and they are expected to enhance the use of natural products as drugs.

Background : Acanthopanax sessiliflorus (Rupr. et Maxim) Seem, belonging to the Araliaceae family, is widely distributed in Korea, China, and Japan. The plants belonging to Acanthopanax species are traditionally used in Korea as anti-rheumatoid arthritis, anti-inflammatory and anti-diabetic drugs and are recognized to have ginseng-like activities. A simple and sensitive high-performance liquid chromatographic (HPLC) method was developed and validated for independent analysis of major compounds and chlorogenic acid in A. sessiliflorus fruits. Chlorogenic acid was reported that prevent cancer and cardiovascular disease in vivo. Also, it has antioxidant effect in vitro test. In the previous experiment, chlorogenic acid were found in A. sessiliflorus fruits. This study was performed to identification of the major compounds and investigate the method validation for the determination of chlorogenic acid in A. sessiliflorus fruits. Methods and Results : Three major compounds were recorded on a Varian Unity Inova AS-400 FT-NMR spectrometer and analyzed by the new HPLC analysis method. HPLC analysis was carried out using an Waters e2695 and PDA detector. The new analyasis method was validated by the measurement of intra-day, inter-day precision, accuracy, limit of detection (LOD, S/N=3), and limit of quantification (LOQ, S/N=10) of chlorogenic acid. The results showed that the correlation coefficient (R2) for the calibration curves of chlorogenic acid was 0.997 in terms of linearity. The limit of detection (LOD) and limit of quantification (LOQ) were 0.565 ㎍/ml and 2.88 ㎍/ml, respectively. There was no interfering peak observed each other and HPLC system was suitable for analysis showing goodness of peak and high precision. Conclusion : This method is suitable to detect and quantify major compounds in A. sessiliflorus fruits. Furthermore, the result will be applied to establish chlorogenic acid as an standard compound for A. sessiliflorus fruits.

Background : Cynanchum wilfordii and Cynanchum auriculatum belong to the Asclepiadaceae family and appear morphologically similar. In order to discriminate them, it is needed to find the presence of sap and the leaf shapes: C. auriculatum has a blade ovate leaf comparing to C. wilfordii. However, in the herbal medicine market, they have been handled as cut and dried roots. Due to their similar morphology, it is limited to distinguish the roots of C. wilfordii and C. auriculatum. Recently in Korea, it has been a critical issue to misuse these two roots in the herbal market and food industry. Thus, it is required to establish a robust tool for the discrimination and quality control of them. Methods and Results : To separation and characterization of flavor compounds, C. wilfordii and C. auriculatum samples were analyzed by head space solid phase micro extraction (HSS) fiber coupled with gas chromatography-mass spectrometry using Rtx-5MS (30 m x 0.25 mm x 0.25 μm) column. As a result, We have identified compounds of a few hundred in aliphatic aldehydes and aliphatic alcohols, alkenes and acids, aromatic compounds, aromatic compounds containing nitrogen & sulfur, etc,. In particular, The aliphatic and aromatic compounds had been clearly separated on the second dimensional direction by using two-dimensional GC. Conclusion : The volatile flavor compounds of C. wilfordii and C. auriculatum could easily analyzed without pre-treatment with improved resolution and sensitivity using HSS-GCxGC-TOFMS. We have identified compounds of a few hundred in C. wilfordii and C. auriculatum sample. And It was more accurately qualitative confirmed with separation of GCxGC and TSD. We have confirmed the PCA and PLS-DA Plot that was classified depending on C. wilfordii and C. auriculatum through multivariate statistical analysis of the identified flavor compounds.

Background : Ginseng has been commonly used as a traditional oriental medicine for its wide spectrum of medicinal properties, including anti-inflammatory, antitumorigenic, adaptogenic and anti-aging properties. 20(S)-Protopanaxadiol (PPD), a intestinal metabolite of ginsenosides, is one of the active ingredients in ginseng. In this study, we have found inflammation-related genes regulated by 20(S)-PPD in mouse bone marrow-derived macrophage (BMDM) to elucidate the role of 20(S)-PPD in inflammatory signaling pathways. Methods and Results : We examined cell viability of BMDM cells after treatment of 20(S)-PPD and found that 20(S)-PPD has no cytotoxicity up to 10 uM. BMDM cells treated with none or 10uM 20(S)-PPD were used for RNA extraction and microarray analysis. It was found that 2 inflammation-related genes are upregulated and 4 genes are downregulated by 20(S)-PPD. Conclusion : These results can give clues to elucidate the role of 20(S)-PPD in inflammatory signaling pathways.

Background : Recently, ginseng (Panax ginseng C.A. Meyer.) berry has been used as a health-promoting supplements. Also, Mulberries (Morus alba L.) fruit have been used in traditional herbal medicine to treat and prevent diabetes. In this study, we measured the cytotoxicity after fermentation of the extracts in Panax Ginseng Berry and Mulberry Fruit. Methods and Results : The extracts were prepared by decoction for 3 hours in distilled water (100 g/L). The dried extract was then dissolved in phosphate-buffered saline (PBS) in preparation for use. Cell viability was examined by an MTT assay. RAW 264.7 cells were seeded at 1 × 104/mL densities in 96-well plates. Each grouping had a non-treated group as the control. The extracts were added to each well and incubated for 24 hours at 37°C and 5% CO2. The MTT solutions (5 mg/mL) were added to each well, and the cells were cultured for another 2 hours. The supernatant was then discarded, and 100 μL of dimethyl sulfoxide was added to each well. The optical density was read at 540 nm. Conclusion : Probiotics and prebiotics modulate the composition of human and domestic animal gut microbiota. The beneficial effects may result from suppression of harmful microorganisms or stimulation of organisms which contribute in a positive way to the nutrition and health of human and domestic animal. Recently, fermentation using microorganisms for the production of more effective compounds has been extensively studied. In particular, the novel pharmacological effects of a new compound generated by fermentation have been reported. Some previous studies have demonstrated that Fermented herbal medicine extract showed better bioactivity than normal herbal Plants extract when used at the same concentration.

Background: The public has increasing concerns about herbal crops owing to insufficient information on biological hazards such as foodborne pathogens. Therefore, the objective of this study is the development of a herbal crop quality control system through monitoring with biological hazard analysis. Today, it is estimated that millions of people become ill every year from food contamination. The public demands agricultural products of stable and consistent quality. Governments have the responsibility of establishing the standards, legislation and enforcement programs necessary to control food quality and safety. However, research on the biosafety of herbal crop products is still insufficient. Therefore, the implementation of monitoring systems with high standards is critical for public safety. Methods and Results: In this study, we collected 52 samples of herbal crop products, and conducted both quantitative and qualitative biological hazard analysis. With biological hazard analysis, aerobic bacteria, Staphylococcus aureus, Salmonella spp., Escherichia coli, Coliforms, and Listeria spp. could be detected. Conclusions: Herbal crops were found to be contaminated with aerobic bacteria at 3.69 ± 0.32 log CFU/g. Staphylococcus aureus, Salmonella spp., Escherichia coli, Coliforms, and Listeria spp. were not detected in any of the samples. This research suggests that continuous monitoring of biological hazards is required to improve the quality of herbal crops.