• Natural Product Sciences
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• 제21권2호
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• pp.104-110
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• 2015

The activities on the inhibition of NO on LPS-induced RAW 264.7 macrophages were investigated in this work. A simple and sensitive method has been developed and validated for fingerprinting analysis of leaves of Acanthopanax gracilistylus W.W. Smith (AGS). The cytotoxicity and inhibition of NO on LPS-induced RAW 264.7 cells of the extract and triterpenoids were determined. Optimal conditions of HPLC analysis were established as follows. The separation was performed with an ODS-C₁₈ column at $30^{\circ}C$, the detected wavelength was 210 nm, the flow rate was 1 mL/min, and the mobile phase consisted of acetonitrile (0.05% phosphoric acid)-0.05% phosphoric acid solution with gradient elution. Our results showed that impressic acid and acankoreaogenin was more effective on the inhibition of NO than the methanol extract and other compounds. There were seventeen peaks coexisted with similarities above 0.95 and nine lupane-triterpenoids including acankoreaogenin and impressic acid detected and identified. The result of anti-inflammatory activities provides a potential explanation for the use of AGS leaves as a herbal medicine in the treatment of inflammatory diseases. Our results also show that acankoreanogenin and impressic acid may be potentially useful in developing new anti-inflammatory agents. In addition, the fingerprint chromatography clearly illustrated and confirmed the material basis for the anti-inflammatory activities of this plant.

• 대한한의학방제학회지
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• 제28권4호
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• pp.327-337
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• 2020

Objective : Citri Unshius Pericarpium is the dried peel of mature fruit of Citrus unshiu Markovich and has been used to treat indigestion, vomiting, and removal of phlegm. This study investigated the hepatoprotective and antioxidant effect of CEE (Ethanol extract of Citri Unshius Pericarpium) in cadmium (CdCl₂)-treated HepG2 cells. Methods : Component analysis of Citri Unshius Pericarpium was analyzed by UPLC with C₁₈ column. Cell viability was determined by MTT assay. The enzyme activity of superoxide dismutase (SOD) and the level of reactive oxygen species (ROS) and reduced glutathione (GSH) were analyzed using commercially available kits. Results : Cadmium caused severe HepG2 cell death. Cadmium also increased ROS production, consistent with depletion of GSH and inhibition of the SOD enzyme. However, CEE treatment reduced cell death and relieved oxidative stress caused by cadmium toxicity. CEE lowered ROS levels and improved depletion of GSH levels. CEE also enhanced the enzymatic activity of SOD. In component analysis, hesperidin was the most abundant of the five marker compounds (Narigenin, Narigin, Narirutin, Hesperidin and Hesperidin), which assumes that hesperidin partially contributed to the antioxidant activity of CEE. Conclusion : These results suggested that CEE could be a potential substance to solve heavy metal-related health problems. In particular, inhibition of oxidative stress by CEE can be a way to treat liver damage caused by cadmium.

• 생약학회지
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• 제40권4호
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• pp.298-302
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• 2009

Samultang is one of traditional medicine composed of Paeonia lactiflora, Angelica gigas, Rehmannia glutinosa and Cnidium officinale. To develop simultaneous determination of paeoniflorin, decursin and 5-HMF in Samultang, a high performance liquid chromatography with diode array detector was used. To separate three marker components, Dionex $C_{18}$ column (5 ${\mu}m$, 120 ${\AA}$, 4.6 mm${\times}$150 mm) was used with a gradient elution system of water and methanol. UV wavelength of detector set at 230 nm and 280 nm. This method was validated by linearity, precision test and recovery test. Calibration curves of three standard components were showed good linear regression ($R^2$>0.9973). LOD and LOQ ranged from 0.08 ${\mu}g$/ml to 0.38 ${\mu}g$/ml and 0.25 ${\mu}g$/ml to 1.16 ${\mu}g$/ml, respectively. The relative standard deviations (RSDs) of data of the inter-day and intra-day experiments were less than 0.54% and 0.89%, respectively. The measured results of recovery test were varied from 93.36 to 107.79 with RSD values 0.01~1.45%. The established method was applied for separation of bio-conversion Samultang sample and compared with control sample.

• Environmental Analysis Health and Toxicology
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• 제29권
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• pp.18.1-18.9
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• 2014

Objectives The purpose of this study was to determine a separation method for each arsenic metabolite in urine by using a high performance liquid chromatography (HPLC)-inductively coupled plasma-mass spectrometer (ICP-MS). Methods Separation of the arsenic metabolites was conducted in urine by using a polymeric anion-exchange (Hamilton PRP X-100, $4.6mm{\times}150mm$, $5{\mu}m$) column on Agilent Technologies 1260 Infinity LC system coupled to Agilent Technologies 7700 series ICP/MS equipment using argon as the plasma gas. Results All five important arsenic metabolites in urine were separated within 16 minutes in the order of arsenobetaine, arsenite, dimethylarsinate, monomethylarsonate and arsenate with detection limits ranging from 0.15 to $0.27{\mu}g/L$ ($40{\mu}L$ injection). We used G-EQUAS No. 52, the German external quality assessment scheme and standard reference material 2669, National Institute of Standard and Technology, to validate our analyses. Conclusions The method for separation of arsenic metabolites in urine was established by using HPLC-ICP-MS. This method contributes to the evaluation of arsenic exposure, health effect assessment and other bio-monitoring studies for arsenic exposure in South Korea.

• 한국식품과학회지
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• 제48권4호
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• pp.335-340
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• 2016

본 연구에서는 다양한 식품으로부터 식중독 세균의 DNA를 추출하는 효율을 비교 하였다. 사용된 DNA 추출 방법은 TaKaRa Kit를 이용하는 방법, PrepMan reagent를 이용하는 방법, boiling method, PEG를 이용한 alkaline method가 사용되었다. 비용절감이나 시간절약 면에서 boiling method나 PrepMan method도 고려할 수 있지만, column kit를 이용하는 TaKaRa kit가 효율적이라고 판단되었다. 또한 정성 시험법에서 적은 양의 균을 검출하기 위해 증균배양을 거치게 되는데 이때 사용되는 증균배지의 성분이 DNA 추출 후에도 잔류하여 saline과 비교하였을 때 DNA 추출효율이 낮은 결과를 나타내었다. 따라서 증균배지의 성분을 제거한 뒤 DNA를 추출하는 것이 PCR의 효율을 높일 수 있을 것으로 예상된다. 정량 시험법에서는 증균과정을 거치지 않아 균을 검출할 때 DNA의 추출 효율이 중요하기 때문에 DNA 추출 효율을 높이기 위한 가장 좋은 버퍼를 선정하고자 하였다. 그 결과 E. coli O157:H7에서는 saline이, S. aureus에서는 증류수를 버퍼로 사용했을 때 DNA 추출 효율이 가장 높은 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제12권2호
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• pp.301-305
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• 2002

The gene (ppaT) encoding Thermus caldophilus GK24 pyrophosphatase (Tca pyrophosphatase) was cloned and sequenced. The gene was found to contain an open reading frame encoding 175 amino acids with a calculated mass of 19,155 Da. The ppaT gene was expressed under the control of the tac promoter in Escherichia coli. The recombinant Tca pyrophosphatase was purified 21.4-fold with $56\%$ yield and specific activity of 25.7 U $mg^-1$, following a combination of heating (to denature the E. coli proteins) and one step of DEAE-Sephacel column chromatography. The native enzyme was found to have an approximate molecular mass of 110,000 Da and consisted of six subunits. The enzyme exhibited maximal activity at pH of 8.0-8.5 and was stable at $80-90^{\circ}C$. A divalent cation was absolutely required for the enzyme activity, with $Mg^2+$. being the most effective.

• Journal of Microbiology and Biotechnology
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• 제12권6호
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• pp.972-978
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• 2002

Streptomyces sp. AD001 is a Gram-positive soil actinomycetes secreting an uncharacterized 2,4-dichlorophenol (DCP) oxidizing enzyme, whose activity is similar to the previously known Actinomycetes lignin-peroxidase (ALiP). This extracellular peroxidase was purified from Streptomyces sp. AD001 as a single protein band on an SDS-PACE by ammonium sulfate fractionation, Q-sepharose, concanavalin A, and Bio-Gel HTP column chromatographies. The molecular mass of the purified peroxidase was determined by SDS-PAGE to be 45.2 kDa, and 49.7 kDa with MALDI-TOF-MS, respectively. The highest level of peroxidase activity was observed at pH 7.5 and $30^{\circ}C$. The amino terminal sequence of the purified peroxidase (G-E-P-E-E-G-N-V-D-G-T-L) showed no significant homologies to my known proteins, suggesting that Streptomyces sp. AD001 may secrete a novel kind of bacterial peroxidase Initial rate kinetic data of the 2,4-DCP oxidation were best modeled with a random-binding bireactant system.

• Ocean and Polar Research
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• 제30권3호
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• pp.225-238
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• 2008

In order to investigate the effects of intertidal sediments on the nutrient cycle in coastal environments, the benthic fluxes of ammonium, nitrate, nitrite, phosphate, and silicate at two stations on the intertidal flat of Keunso Bay were determined during each season. The efflux of ammonium was observed at S1 and resulted from the diffusion of remineralized ammonium and acceleration caused by the bioirrigation of macrofauna. The influx of ammonium at S2 was probably due to nitrification in the water column. The influx of nitrate was observed at both stations during all seasons, indicating that the nitrate in the pore water was removed by denitrification. Vigorous bioirrigation led to the efflux of dissolved inorganic nitrogen (DIN) at S1, whereas the influx of DIN at S2 was predominantly caused by denitrification. Contrary to the diffusive and bio-irrigated release of remineralized phosphate from the sediment at S1, the influx of phosphate was observed at S2, which may be attributable to adsorption onto iron oxides in the aerobic sediment layer. Silicate, which is produced by the dissolution of siliceous material, was mostly released from the sediment by molecular diffusion and bioirrigation. However, the influx of silicate was observed at S2 during spring and winter, which was ascribed to adsorption by particulate matter or assimilation by benthic microphytes. The annual fluxes of DIN were 328 mmol $m^{-2}yr^{-1}$ at S1 and -435 mmol $m^{-2}yr^{-1}$ at S2. The annual fluxes of phosphate were negative at both sites (-2.8 mmol $m^{-2}yr^{-1}$ at S1 and -28.9 mmol $m^{-2}yr^{-1}$ at S2), whereas the annual fluxes of silicate were positive at both sites (843 mmol $m^{-2}yr^{-1}$ at S1 and 243 mmol $m^{-2}yr^{-1}$ at S2).

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.274-279
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• 2015

Receptor activator of nuclear factor-kappa B ligand (RANKL) is a critical factor in osteoclastogenesis. It makes osteoclasts differentiate and multinucleate in bone remodeling. In the present study, RANKL was expressed as a soluble maltose binding protein (MBP)-fusion protein using the Escherichia coli maltose binding domain tag system (pMAL) expression vector system. The host cell E. coli DH5α was cultured and induced by isopropyl β-_D-1-thiogalactopyranoside for rRANKL expression. Cells were disrupted by sonication to collect soluble MBP-fused rRANKL. The MBP-fusion rRANKL was purified with MBP Trap affinity chromatography and treated with Tobacco Etch Virus nuclear inclusion endopeptidase (TEV protease) to remove the MBP fusion protein. Dialysis was then carried out to remove binding maltose from the cleaved rRANKL solution. The cleaved rRANKL was purified with a second MBP Trap affinity chromatography to separate unsevered MBP-fusion rRANKL and cleaved MBP fusion protein. The purified rRANKL was shown to have biological activity by performing in vitro cell tests. In conclusion, biologically active rRANKL was successfully purified by a simple two-step chromatography purification process with one column.

• KSBB Journal
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• 제27권3호
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• pp.151-156
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• 2012

Lactic acid is an important product arising from the anaerobic fermentation by lactic acid bacteria (LAB). It is used in the pharmaceutical, cosmetic, chemical, and food industries as well as for biodegradable polymer and green solvent production. The poly lactic acid (PLA) is an important material for bio-plastic manufacturing process. For PLA production by new LAB, we screened LAB isolates from shellfish. A total of 28 LAB were isolated from various shellfishes. They were all Gram positive, oxidase and catalase negative. Based on API 50CHL kit, 7 strains among the 28 isolates were identified as Lactobacillus plantarum, 6 strains as Lactobacillus delbrueckii, 5 strains as Leuconostoc mesenteroides, 3 strains as Lactobacillus brevis, 2 strains as Lactococcus lactis, 1 strain as Lactobacillus salivarius, 1 strain as Lactobacillus paracasei, 1 strain as Lactobacillus pentosus, 1 strain as Lactobacillus fermentum and 1 strain as Pediococcus pentosaceu. Also, we examined the amount of total lactic acid produced by these new strains by HPLC analysis with Chiralpak MA column. One strain E-3 from Mytilus edulis was indentified as Lactobacillus plantarum and found to produce 20.0 g/L of D-form lactic acid from 20 g/L of dextrose. Further studies are underway to increase the D-lactic acid production by E-3.