• Title/Summary/Keyword: TLC analysis

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Effectiveness of Bioremediation on Oil-Contaminated Sand in Intertidal Zone

  • Oh, Young-Sook;Sim, Doo-Suep;Kim, Sang-Jin
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.437-443
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    • 2003
  • Bioremediation technologies were applied to experimental microcosms, simulating an oil spill in a lower intertidal area. Three treatments (oil only, oil plus nutrients, and oil plus nutrients and microbial inocula) were applied, and each microcosm was repeatedly filled and eluted with seawater every 12 h to simulate tidal cycles. To minimize washing-out of the inoculum by the tidal cycles, microbial cells were primarily immobilized on diatomaceous earth before they were applied to the oiled sand. Oil degradation was monitored by gravimetric measurements, thin layer chromatography/flame ionization detector (TLC/FID) analysis, and gas chromatography (GC) analysis, and the loss of oil content was normalized to sand mass or nor-hopane. When the data were normalized to sand mass, no consistent differences were detected between nutrient-amended and nutrient/inoculum-amended microcosms, although both differed from the oil-only microcosm in respect of oil removal rate by a factor of 4 to 14. However, the data relative to nor-hopane showed a significant treatment difference between the nutrient-amended and nutrient/inoculum-treated microcosms, especially in the early phase of the treatment. The accelerating effect of inoculum treatment has hardly been reported in studies of oil bioremediation in the Tower intertidal area. The inoculum immobilized on diatomaceous earth seemed to be a very effective formulation for retaining microbial cells in association with the sand. Results of this study also suggest that interpretation of the effectiveness of bioremediation could be dependent on the selection of monitoring methods, and consequently the application of various analytical methods in combination could be a solution to overcome the limitations of oil bioremediation monitoring.

Comparison of Analytical Methods for Saikosaponins in Bupleurum falcatum L. (자호(紫胡) 사이코사포닌 정량분석방법(定量分析方法) 비교(比較))

  • Kim, Kwan-Su;Lee, Seoung-Tack;Seong, Nak-Sui;Lee, Jung-Il;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.3
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    • pp.226-232
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    • 1995
  • Extraction methods and instrumental analytical conditions were compared to establish a fast and appropriate analytical method to determine saikosaponins in Bupleurum falcatum. Using HPLC, analysis of diene-saikosaponin treated with 2% acid was faster than that of saikosaponin itself. Among various extraction methods, extraction by standing in methanol at room temperature showed highest efficieny, and extraction with boiling methanol was shorter in analytical time and showed good chromatogram. And we could analyze many samples faster using HPTLC but the analytical accuracy was low. In extraction and analysis of saikosaponins, extraction with boiling methanol and acidic treatment was fast and easy analytical method. And for selecting useful lines in component breeding, we think TLC method was better.

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An Isolation of Crude Saponin from Red-Ginseng Efflux by Diaion HP-20 Resin Adsorption Method (홍삼유출액으로부터 Diaion HP-20 수지 흡착법에 의한 조사포닌의 분리)

  • 곽이성;경종수;김시관;위재준
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.1
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    • pp.1-5
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    • 2001
  • This study was carried out to isolate saponin compounds from red-ginseng efflux, which was produced during the industrial processing of red-ginseng from fresh ginseng. We isolated crude saponin from the efflux extract (moisture content 35.0%) by using Diaion HP-20 adsorption method. Non-saponin fraction, which was adsorbed on Diaion HP-20 resin, was removed by eluating with $H_{2}O$ and 25% spirit. Then crude saponin was eluated with 95% spirit, continuously. Saponin in the eluated fractions was confirmed by TLC analysis. Crude saponin isolated from red ginseng efflux extract contained 12.10% of saponin. whereas those of white ginseng and red-ginseng were 3.30 and 3.39%, respectively. Ginsenoside contents showed the highest contents kin crude saponin from red ginseng efflux extract. Expacilly, the ginsenoside-$Rb_{1}$ and Re showed the highest contents in red-ginseng efflux extract when compared with those of white ginseng and red ginseng crude saponins. And the other ginsenosides except ginsenoside-$Rb_{1}$ and -Re also showed the highest contents in red ginseng efflux extract. However, the ratio of PD saponin (Panaxadiol saponin: $Rb_{1}+Rb_{2}$+Rc+Rd) to PT saponin (panaxatriol: $Re+Rg_{1}$) showed almost the same level when compared with those of ginseng saponin fractions. Ratio of PD/PT from red ginseng efflux extract was 1.99. Ratios of PD/PT from white ginseng and red ginseng were 1.85 and 1.84, respectively. Saponin purity, which was calculated by ratio percent of total ginsenoside to curde saponin content, was 45.90%. In case of white ginseng and red ginseng, the purities were 35.50 and 36.00%, respectively. However, by PHLC analysis, we confirmed that crude saponin isolated from red ginsengs. It suggested that crude saponin isolated from red ginseng ellux also would be useful component as ginseng saponins.

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JAK-2 V617F Mutational Analysis in Primary Idiopathic Myelofibrosis: Experience from Southern Pakistan

  • Sultan, Sadia;Irfan, Syed Mohammed
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7889-7892
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    • 2015
  • Background: Primary idiopathic myelofibrosis (PMF) is a clonal Ph-chromosome negative myeloproliferative neoplasm characterized by dysregulated kinase signaling and release of abnormal cytokines. In the recent past, following JAK2 V617F mutation invention, important revolution has been made in the molecular diagnostic biology of this disease. The rational of this study was to determine the mutational status of JAK2 V617F in Pakistan patients with PMF. Materials and Methods: In this cross sectional study, 20 patients with PMF were enrolled from January 2011 to December 2014. Diagnosis was based on WHO criteria for PMF. All patients were screened for G-T point mutation (V617F) in the JAK2 gene on chromosome 9 by allele specific PCR. Results: The mean age was $57.9{\pm}16.5years$. The male to female ratio was 3:1. The frequency of JAK2 V617F positivity in our PMF patients was found to be 55%. Positive correlations of JAK2 V617F mutation were established with high TLC count, raised LDH and marked splenomegaly (P<0.05). No correlation of JAK2 V617F could be established with age and gender (P>0.05). Conclusions: The JAK2 V617F mutation frequency in our PMF patients was similar to those reported previously. In our hands JAK2 V617F mutated patients expressed an aggressive disease phenotype. Screening for the mutation in all suspected PMF cases could be beneficial in differentiating patients with reactive and clonal marrow fibrosis.

Plant growth promotion effect of Klebsiella michiganensis Jopap-1 isolated from button mushroom bed (양송이배지로부터 분리한 Klebsiella michiganensis Jopap-1의 식물생장촉진효과)

  • Kim, Ye-Seul;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.218-224
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    • 2018
  • An auxin-producing bacterium, Klebsiella michiganensis Jopap-1, was isolated from a button mushroom bed in Buyeo-Gun, Chungcheongnam-Do. The strain Jopap-1 was classified as a novel strain of K. michiganensis based on a chemotaxonomic and phylogenetic analysis. The isolated K. michiganensis Jopap-1 was confirmed to produce indole-3-acetic acid (IAA), which is one of auxin hormones by TLC and HPLC analyses. The maximum concentration of IAA ($96.05mg\;L^{-1}$) was detected in the culture broth incubated in R2A medium containing 0.1% L-tryptophan for 48 h at $35^{\circ}C$ by HPLC quantity analysis. A negative relationship between IAA production and pH variation was estimated to show that the increase of IAA caused acidic pH in the culture. The effect of the supplement on L-tryptophan (precursor of IAA) production was observed to be highest at 0.1% concentration, but was significantly lowered above a concentration of 0.2%. To investigate the growth-promoting effects on the crops, the culture broth of E. michiganensis Jopap-1 was infected to water cultures and seed pots of mung bean and lettuce. Consequently, the adventitious root induction and root growth of mung bean and lettuce were observed to be 2.1 and 1.8 times higher than those of the control.

Biosynthesis of New Oligosaccharides via Acceptor Reaction using Fructosyloligosaccharide as an Acceptor (Fructosyloligosaccharide를 Acceptor 반응의 기질로 사용한 새로운 올리고당의 생합성)

  • 이찬용;이충환
    • Korean Journal of Microbiology
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    • v.35 no.2
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    • pp.146-152
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    • 1999
  • Acceplor reaction of glucosyltr~ansferase of Streptococcus ,SO~~-~IZLIS with f ~ ~ t o ~ y l o l i g o ~ a ~ ~ h a r i d e ~ was studied for the biosynthesis of novel olgisaccharides. First, bacilracin resistant mutants were selected by mutagenesis of Streptococcus sobrimis ATCC27351. Among these mutants 4 strains were selected by resistance to bacitracin and increase of glucosyltransferase. Acceptor reaction of maltose was analyzed by TLC and image analysis. There were differences in the specificity of the acceptor reaclion by Ule glucosylumsferase between mother strain (Streptococcus sobrinus ATCC2735) and bacitracin resistant mutants (Streptococcus sobrinus BR24C, Strepfococcus sobrinus CH-5). Molher strain did ilot show an acceptor reaction with fructosyloligosaccharides such as 1-keqtose and nystose. Acceptor reaction products of turailose and 1-kestose with glucosyltransferase (GW-S) of Streptococcus sobrini~s BR24C were TEX>\6^{3}$-$\alpha$-D-glucopyranosyl \3^{2}$-O-$\alpha$-D-fructose (glucose-fructose-glucose) and \6^{4}$-$\alpha$-D-glucopyranosyl \1^{3}$-$\beta$-D-~-h~ctofuranos~~ sucrose (glucose-glucosefructose- fructose). respectively These are novel oligosaccharides which can be produced only by enzymatic reaction.

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A Study on Sugars in Korean Sweet Rice Drink "Sikhye"(III) -Commercial Sikhye- (식혜의 이소말토올리고당에 관한 연구(III) -시판식혜-)

  • 안용근
    • The Korean Journal of Food And Nutrition
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    • v.10 no.1
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    • pp.92-96
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    • 1997
  • A Korean commercial sweet rice drink "Sikhye" showed sucrose, fructose, glucose, maltose, limit dextrin and various size of maltooligosaccharides in HPLC and TLC analysis. Commercial Sikhye was found to contain 0.09% of limit dextrin and 0.2% of rice residue. Limit dextrin in commercial Sikhye showed both signal of $\alpha$-1,4- and $\alpha$-1,6-glucosidic linkage with its estimation ratio of 15:1 by 1H-NMR analysis. This limit dextrin was hydrolyzed to produce various size of maltooligosaccarides with more longer chain than that of traditional Sikhye by pullulanase. Limit dextrin was digested wit enzymes(30units/ml) of $\alpha$-amylase, $\alpha$-glucosidase and glucoamylase from Aspergillus awamori, sweet potato $\beta$-amylase and human salivary $\alpha$-amylase at 37$^{\circ}C$ for 1 hour, respectively. Hydrolysis rates of these amylases on it were higher than in case of traditional sikhye. $\alpha$-Glucosidase plus human salivary $\alpha$-amylase hydrolyzed it to 61.3%. Hydrolysis rates of these amylases on rice residue were lower than that of traditional Sikye. These results suggest that limit dextrin in commercial Sikhye is less effective than isomaltooligosaccharides in traditional Sikhye as a growth factor for Bifidobacterium while rice residue in commercial Sikhye is more effective than that in traditional Sikhye as dietary fiber.ary fiber.

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Quantitative Analysis of Lysophosphatidic Acid in Human Plasma by Tandem Mass Spectrometry

  • Kim, Ho-Hyun;Yoon, Hye-Ran;Pyo, Dong-Jin
    • Bulletin of the Korean Chemical Society
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    • v.23 no.8
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    • pp.1139-1143
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    • 2002
  • Analysis of lysophosphatidic acids (LPAs) is of clinical importance as they can serve a potential marker for ovarian and other gynecological cancers and obesity. It is critically important to develop a highly sensitive and specific method for the early detection of gynecological cancers to improve the overall outcome of this disease. We have established a novel quantification method of LPAs in human plasma by negative ionization tandem mass spectrometry (MS-MS) using multiple reaction monitoring (MRM) mode without the conventional TLC step. Protein-bound lipids, LPAs in plasma were extracted with methanol : chloroform (2:1) containing LPA C14:0 as an internal standard under acidic condition. Following back extraction with chloroform and water, the centrifuged lower phase was evaporated and reconstituted in methanol. The reconstituted solution was directly injected into electrospray source of MS/MS. For MRM mode, Q1 ions selected were m/z 409, 433, 435, 437 and 457 which corresponds to molecular mass [M-H]- of C16:0, C18:2, C18:1, C18:0 and C20:4 LPA, respectively. Q2 ions selected for MRM were m/z 79, phosphoryl product. Using MS/MS with MRM mode, all the species of LPAs were completely separated from plasma matrix without severe interferences. This method allowed simultaneous detection and quantification of different species of LPAs in a plasma over a linear dynamic range of 0.01-25 ㎛olL-1 . The detection limit of the method was 0.3 pmol/mL, with a correlation coefficient of 0.9983 in most LPAs analyzed. When applied to the plasmas of normal and gynecological cancer patients, this new method differentiated two different groups by way of total LPA level.

Effects of pH and High Temperature Treatment on the Changes of Major Ginsenosides Composition in Korean Red Ginseng Water Extract (pH 및 고온 열처리가 홍삼물추출물의 주종 사포닌 성분변화에 미치는 영향)

  • Choi, Keum-Hee;Kwak, Yi-Seong;Rhee, Man-Hee;Hwang, Mi-Sun;Kim, Seok-Chang;Park, Chae-Kyu;Han, Gyeong-Ho;Song, Kyung-Bin
    • Journal of Ginseng Research
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    • v.32 no.2
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    • pp.127-134
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    • 2008
  • This study was carried out to investigate the changes of ginsenosides composition in Korean red ginseng water extract (RGWE) after heated with high temperatures above $100^{\circ}C$. RGWEs were adjusted with pH 3.0, pH 7.0 and pH 10.0, respectively, and then heated at 100,110 and $120^{\circ}C$ for 30 minutes by using autoclave. Total ginsenosides of RGWE treated with heating showed decreasing tendency when compared with control. By TLC analysis, decreasing effect of ginsenosides in RGWE were significantly observed in the acidic condition of pH 3.0, particulary. By HPLC analysis, total ginsenoside of control showed 1.89%, while those of RGWE treated with 100, 110 and $120^{\circ}C$ showed 1.22, 1.05 and 0.97%, respectively. The ratio of protopanaxadiol (PD) to protopanaxatriol (Pr) saponins in control was 1.89, while that of PD/PT in treated RGWEs were level of 1.33 to 1.47. By the result of decreased ratio of PD/PT in RGWE, it was considered that PD type saponin such as ginsenoside$-Rb_{1}$, $-Rb_{1}$, -Rc and -Rd was more unstable than PT type saponin such as ginsenoside-Re and Rg against high temperature heating above $100^{\circ}C$.

Biotransformation of natural polyacetylene in red ginseng by Chaetomium globosum

  • Wang, Bang-Yan;Yang, Xue-Qiong;Hu, Ming;Shi, Li-Jiao;Yin, Hai-Yue;Wu, Ya-Mei;Yang, Ya-Bin;Zhou, Hao;Ding, Zhong-Tao
    • Journal of Ginseng Research
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    • v.44 no.6
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    • pp.770-774
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    • 2020
  • Background: Fermentation has been shown to improve the biological properties of plants and herbs. Specifically, fermentation causes decomposition and/or biotransformation of active metabolites into high-value products. Polyacetylenes are a class of polyketides with a pleiotropic profile of bioactivity. Methods: Column chromatography was used to isolate compounds, and extensive NMR experiments were used to determine their structures. The transformation of polyacetylene in red ginseng (RG) and the production of cazaldehyde B induced by the extract of RG were identified by TLC and HPLC analyses. Results: A new metabolite was isolated from RG fermented by Chaetomium globosum, and this new metabolite can be obtained by the biotransformation of polyacetylene in RG. Panaxytriol was found to exhibit the highest antifungal activity against C. globosum compared with other major ingredients in RG. The fungus C. globosum cultured in RG extract can metabolize panaxytriol to Metabolite A to survive, with no antifungal activity against itself. Metabolites A and B showed obvious inhibition against NO production, with ratios of 42.75 ± 1.60 and 63.95 ± 1.45% at 50 µM, respectively. A higher inhibitory rate on NO production was observed for Metabolite B than for a positive drug. Conclusion: Metabolite A is a rare example of natural polyacetylene biotransformation by microbial fermentation. This biotransformation only occurred in fermented RG. The extract of RG also stimulated the production of a new natural product, cazaldehyde B, from C. globosum. The lactone in Metabolite A can decrease the cytotoxicity, which was deemed to be the intrinsic activity of polyacetylene in ginseng.