• Food Science and Biotechnology
• /
• v.15 no.3
• /
• pp.482-484
• /
• 2006

The antioxidant activity of Bacillus polyfermenticus SCD was studied by partially purified culture extracts using various methods: ammonium sulfate precipitation, adsorption to Diaion HP-20 columns using polar solvents, and extraction using non-polar solvents. The 1,1-diphenyl-2-picyrylhydrazyl (DPPH) radical scavenging activity of these partially purified fractions was then investigated. The precipitate isolated using 75%-saturated ammonium sulfate was shown to contain about 77.2% DPPH radical scavenging activity. Using the Diaion HP-20 resin adsorption method, the fraction obtained using 60% ethanol and 60% methanol possessed 76.7 and 89.5% DPPH radical scavenging activity, respectively. Fractions obtained by extracting with the non-polar solvents 80 mg/mL chloroform, 80 mg/mL n-hexane, 80 mg/mL ethyl acetate, and 80 mg/mL butanol contained 68.4, 75.0, 70.7, and 87.5% DPPH radical scavenging activity, respectively. Further study is needed to characterize the antioxidant substance(s) released by B. polyfermenticus SCD cultures.

• KSBB Journal
• /
• v.23 no.4
• /
• pp.329-334
• /
• 2008

The antioxidant activity and the qualitative analysis of Acanthopanax sessiliflorum Seeman were studied by partially purified extract using various methods: extraction by using ethanol solutions and temperatures, and absorption to Diaion HP20 column chromatography using 10%, 20%, 40%, 60% ethanol solutions. Major constituents, chlorogenic acid, caffeic acid, eleutheroside E, was determinated by HPLC method in Acanthopanax sessiliflorum S. 10% and 20% ethanol solutions contain chlorogenic acid (3.020$\pm$0.080%, 20.500$\pm$1.150%, respectively). 40% ethanol solution contains caffeic acid and eleutheroside E (12.270$\pm$0.360%, 1.670$\pm$0.140%, respectively). Diaion HP20 fractions (10%, 20%, 40%, 60% ethanol solutions) showed the scavenging activity of radicals and reactive oxygen species with the $IC_{50}$ values of $81.534{\pm}0.992{\mu}g/ml$, $1.748{\pm}0.098{\mu}g/ml$, $11.487{\pm}1.768{\mu}g/ml$, $21.960{\pm}0.547{\mu}g/ml$ against 1,1-diphenyl-2-picrylhydrazly radical and $1713.548{\pm}34.565{\mu}g/ml$, $131.419{\pm}2.235{\mu}g/ml$, $200.681{\pm}2.444{\mu}g/ml$, $757.897{\pm}6.868{\mu}g/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Especially, 20% and 40% ethanol fractions showed more antioxidant activity than dl-$\alpha$-tocopherol. These results suggest that Acanthopanax sessiliflorum S. extract and Diaion HP20 fractions may be useful as a potential source of nutraceutical and cosmetic products.

• Journal of Life Science
• /
• v.7 no.3
• /
• pp.186-191
• /
• 1997

In order to select new anticandiosis agent-producing candidates, we used a modified selection method. Two active fractions designated as S-6279A) and (B) were isolated from the fermentation broth of Streptomycetes sp. S627 which was screened by methanol extraction, diaion HP-20 and silica gel column chromatography (chloroform-me-thanol, and benzene-ethyl acetate), and preparative thin layer chromatography.

• Food Science and Biotechnology
• /
• v.16 no.1
• /
• pp.163-166
• /
• 2007

The cytotoxic effects of partially purified substances from Bacillus polylfermenticus SCD toward a variety tumor cell lines were studied. Cytotoxic activity was determined with regard to the A549 (human lung carcinoma), AGS (human stomach adenocarcinoma), DLD-1 (human colon adenocarcinoma), HEC-1-B (human uterus adenocarcinoma), SW-156 (human kidney carcinoma), and NIH/3T3 (murine normal fibroblast) cell lines using the MTT assay. Cytotoxic substances were partially purified through Diaion HP-20 columns and extracted with methanol or other organic solvents (n-hexane, chloroform, ethylacetate, and butanol). B. polyfermenticus SCD supernatant showed up to 60% inhibition of cell viability fer all five human cancer cell lines tested. When treated with 10 mg/mL of n-hexane, chloroform, ethylacetate, and butanol extract, HEC-1-B cells showed a 25,62,35, and 63% rate of inhibition respectively, and AGS cells showed a 72, 61, 44, and 67% rate of inhibition, respectively. At a concentration of 10 mg/mL, 100% methanol Diaion HP-20 extracts showed inhibition rates of 97.0% toward A-549 cells, 98.1% toward AGS cells, 81.6% toward DLD-1 cells, 83.5% toward HEC-1-B cells, and 92.7% toward SW-156 cells. These results indicate that partially purified fractions from B. polyfermenticus SCD have the potential to inhibit not only colon cancer cells, but also lung, stomach uterus, and kidney cancer cells. Further studies are needed to characterize the cytotoxic substances released in B. polyfermenticus SCD cultures.

• Korean Journal of Pharmacognosy
• /
• v.50 no.3
• /
• pp.198-204
• /
• 2019

A facile and convenient method was developed for the mass production of epigallocatechin gallate (EGCG) rich green tea extract (Er-GTE). The Er-GTE was successfully obtained from the crude water extract of green tea by the combination of two step purification, i.e., a simple adsorption process on the cation exchange resins (Trilite SCR-B) followed by the chromatography with Diaion HP-20 resins. The green tea extract produced by water extraction under $45^{\circ}C$ was subjected to adsorb on the strongly acidic cation exchange resin, Trilite SCR-B. The eluate passed through the resin was reabsorbed on Diaion HP-20 resin, which was subjected to elute with a mixture of water and alcohol by conventional chromatographical manner. The EGCG content in Er-GTE was estimated above 97% by HP-LC analysis and the newly developed method was regarded as the most suitable and appropriate process for the mass production of epigallocatechin gallate rich green tea extract (Er-GTE).

• Korean Journal of Food Science and Technology
• /
• v.33 no.3
• /
• pp.282-287
• /
• 2001

This study was carried out to investigate the difference of ginsenoside compositions in crude ginseng saponins prepared by five different methods including three new methods. Two known methods are hot methanol(MeOH) extraction/n-butanol(n-BuOH) fractionation and hot MeOH extraction/Diaion HP-20 adsorption/MeOH elution. Three new methods are hot MeOH extraction/cation AG 50W $absorption/H_2O$ elution/n-BuOH extraction, cool MeOH extraction/Diaion HP-20 adsorption/MeOH elution and direct extraction with ethyl acetate(EtOAc)/n-BuOH. Analysis of ginsenoside composition in the crude saponins by conventional HPLC/RI(Refractive Index) did not show great difference between methods except EtOAc/n-BuOH method. However, HPLC/ELSD (evaporative light scattering detector) employing gradient mobile phase afforded fine resolution of ginsenoside Rf, $Rg_1$ and $Rh_1$, and great difference of ginsenoside compositions between methods. LC/MS revealed that large amount of prosapogenins were produced during the pass through the cation exchange (AG 50W) column being strongly acidic. Six major ginsenosides such as $Rb_1,w;Rb_2,$ Rc, Rd, Re and $Rg_1$, 5 prosapogenins and one chikusetsusaponin were identified by LC/MS. A newly established HPLC method employing ODS column and gradient mobile phase of $KH_2PO_4/CH_3CN$ revealed that malonyl ginsenosides were detected only in the crude saponin obtained from cool MeOH extraction.

• Korean Journal of Food Science and Technology
• /
• v.33 no.2
• /
• pp.166-172
• /
• 2001

In order to increase ginsenoside content, to reduce chemical change, to shorten extracting procedure, new methods of extraction and fractionation of crude ginseng saponin were established and compared for their chemical composition. Those are hot MeOH extraction/n-BuOH fractionation (BuOH method) and hot MeOH extraction/Diaion HP-20 adsorption/MeOH elution (HP-20 method), which are already known methods, and additional three new methods: hot MeOH extraction/cation AG 50W $adsorption/H_2O$ elution/n-BuOH extraction (AG 50W method), cool MeOH extraction/Diaion HP-20 adsorption/MeOH elution (cool extraction method) and direct extraction with EtOAc/n-BuOH (direct extraction method). AG 50W method provided a crude saponin showing the highest content of ginsenosides of 61.5% and the lowest contents of protein and free amino acids of 0.93% and 0.19%, respectively. The protein content was the highest as 14.18% in the crude saponin by HP-20 method, while free sugar content was the highest as 13.5% by BuOH method, indicating that these are factors that lower the rate of ginsenoside in crude saponins by those methods. On the other hand, it was revealed that AG 50W method produced large amount of prosapogenins during the pass through the cation exchange resin (AG 50W) column being strongly acidic. Crude saponin from direct extraction method showed relatively higher composition of ginsenoside $Rg_1$ and Re. The results suggest that contents and composition of ginsenosides and other chemical components in crude ginseng saponin greatly depend on the condition of the extraction and fractionation.

• Journal of Environmental Health Sciences
• /
• v.31 no.3
• /
• pp.221-226
• /
• 2005

Isoflavone was extracted with various concentration of aqueous methanol using whole hypocotyls as the starting material. Whole hypocotyls were preferred as the raw material because the residue could be easily removed from the solvent after the extraction process. Extraction yield was almost constant at the methanol concentration of $20-80\%$. Most of the isoflavone was extracted within 1 hr, and the extraction yield remained almost constant thereafter. When the concentration of methanol was $80\%$, the content of total solid was reduced due to the reduced extraction of contaminating protein as the result of protein insolubilization. Among resins tested, Diaion HP-20, Amberlite XAD-16, and Amberlite IRC-50 showed the highest capacity to absorb the compound. Open column chromatography with Diaion HP-20 showed that $80\%$ aqueous ethanol was most efficient as the eluting solvent with final recovery of the phytochemical being more than $95\%$. Maximum adsorption of the phytochemical occurred at the acidic pH 2-4. When the spatial velocity was increased to 15 and more, the degree of adsorption was decreased, whereas below spatial velocity of 15, the adsorption capacity of isoflavone to the resin was almost constant. The purity of the isoflavone purified by column chromatography was $78\%$.

• Microbiology and Biotechnology Letters
• /
• v.26 no.6
• /
• pp.558-561
• /
• 1998

In the screening for lipid peroxidation inhibitors from edible mushroom, Agrocybe cylindracea, a bioactive compound AG 8 was isolated. The AG 8 was purified from methanol extract of its fruit body by Diaion HP-20 column chromatography, ethyl acetate extraction, and silica gel column chromatography, consecutively. Based on various NMR studies including $^1$H irradiation and HMBC experiments, the AG 8 was identified as MTA, 5'-deoxy-5'-methylthioadenosine. This compound inhibited lipid peroxidation with an $IC_{50}$/ value of 3.2 $\mu\textrm{g}$/$m\ell$. The MTA was isolated for the first time from basidiomycetes.

• Food Science and Preservation
• /
• v.16 no.5
• /
• pp.754-758
• /
• 2009

To develop soybean cake as a functional food material, the anti-oxidative and cytotoxic activities against human colon cancer cells of crude saponins isolated from 70% (v/v) ethanol extracts of cake were investigated. The Diaion HP-20 adsorption method was used for isolation of crude saponins, which were then eluted with 100% ethanol. The non-saponin fraction was removed by elution with $H_2O$ and 20% (v/v) ethanol. The results of thin layer chromatography (TLC) analysis confirmed that crude saponins were present in the 100% ethanol extract of soybean cake. The hydrogen-donating properties of saponins were more than 60% at a concentration of $1,000\;{\mu}g/mL$. malondialdehyde(MDA) production was $1,200\;{\mu}mol\;MDA/g$ in mouse liver homogenate treated with crude saponins at the concentration of $1,000\;{\mu}g/mL$. This value was lower than that of the control, which was $3,700\;{\mu}mol\;MDA/g$. Saponins inhibited the growth of colon cancer cells in a dose- and time-dependent manner. Saponins also resulted in a decrease in the proportion of cells in the G1 phase of the cell cycle, whereas the cell proportion in G2/M phase was increased with $1,000\;{\mu}g/mL$ saponins. Thus, we conclude that saponins may induce G2/M cell cycle arrest.