• Advances in Traditional Medicine
• /
• v.8 no.3
• /
• pp.228-235
• /
• 2008

This study was conducted to evaluate the estrogen activity of silkworm (Bombyx mori) pupa extracts and their fractions. Powdered samples of freeze-dried silkworm pupa were extracted at room temperature (RT), $40^{\circ}C$, $60^{\circ}C$, $80^{\circ}C$, and $100^{\circ}C$ in water (D.W), chloroform, ethyl acetate, and methanol for 6h and then filtered (0.45 um). The extracts were then freeze-dried. The estrogenic activity of these extracts was then investigated by competition binding assays using estrogen receptor ${\alpha}\;(ER{\alpha})$ and $ER{\beta}$, and by evaluating their effects on the proliferation of the human breast cancer cell line, MCF-7. Among the extracts evaluated, water extracts prepared at RT showed the highest binding affinity to $ER{\alpha}$ ($IC_{50}$, 1.76 ug/ml) and $ER{\beta}$ ($IC_{50}$, 0.07 ug/ml). In addition, MCF-7 cells that were treated with 62.5 ug/ml of the RT extract showed the greatest increase in proliferation (2-fold; 1291.79%) when compared to control cells (659.82%). Next, the water extract that was prepared at RT (sample 1) was dissolved in D.W. and further fractionated using a Dowex 50W - 8X ($H^+$) column. The flow-through and wash were then pooled together and freeze-dried (sample 2). The bound materials were then eluted with 20 mM NaCl, after which they were applied to a Dowex 1X2 - 200 ($Cl^-$) column and washed with D.W. to remove the sodium ions. The eluants were then freeze-dried (sample 3). Of these fractions, sample 2 showed the highest binding affinity to ER{\alpha} ($IC_{50}$, 1.44 ug/ml) and $ER{\beta}$ ($IC_{50}$, 1.18 ug/ml). In addition, MCF-7 cells that were treated with sample 2 (15.6 ug/ml) showed the largest increase in growth (1159.39%) when compared to control cells (525.26%). Taken together, these results suggest that the fraction of the RT water extract of silkworm pupa referred to as sample 2 may be useful as a phytoestrogen.

• YAKHAK HOEJI
• /
• v.29 no.3
• /
• pp.144-151
• /
• 1985

Cathodic stripping voltammetric determination of traces of selenium in plant samples was studied. Stripping peak of selenium (IV) from Cu-Se intermettalic deposit in acidic media containing copper (II) ion is specific, highly sensitive and well defined, is successfully used for the quantitative determination of selenuin down to the level of 1ng/ml. Sample is burnt in a calorimeter bomb under the oxygen pressure of 40atm. and the selenium is absorbed in 0.1M NaOH. After the solution is filtrated, concentrated and acidified with HCl, then passed through a column of cation exchange resin in the $H^{+}$ form(Dowex 50X-8). The column eluate is analyzed for selenium by differential pulse cathodic stripping voltammetric method. Analytical results of selenium for NBS SMR is well agreement with the certified values. Results are given for a series of plant materials.

• Bulletin of the Korean Chemical Society
• /
• v.31 no.12
• /
• pp.3755-3759
• /
• 2010

A on-line SPE (solid phase extraction)-HPLC preconcentration method was developed for the determination of phenolic compounds at trace levels in environmental water sample. XAD-4 and Dowex 1-X8 were used as sorbent in the on-line SPE-HPLC method for the selective enrichment of nine phenolic compounds, which are included in the priority pollutants list of the US EPA. Also alumina prefiltering considerably reduced the amount of interfering peaks due to humic substances that could accumulated due to the preconcentration step and prevent quantification of polar phenolic compounds in environmental water samples. This method was used to determine the phenolic compounds in tap and river water and superiority to the US EPA 625 method in its enrichment factor, pretreatment time, recoveries, and detection limit. The limits of detection were in the range of $0.3-0.9\;{\mu}g/L$ in tap water sample.

• Analytical Science and Technology
• /
• v.13 no.5
• /
• pp.608-615
• /
• 2000

A determination method of trace nickel and cobalt in water samples was studied and developed by adsorbing their complexes on ion exchange resin suspension. The analytical ions were formed as complexes with a ligand of APDC (ammonium pyrrolidinedithiocarbamate) and adsorbed on anion exchange resin of Dowex 2-X8. After the suspension was filtered out with membrane filter, the complexes were dissolved in HCl solution by an ultrasonic vibrator for ET-AAS determination. Several conditions were optimized as followings. pH of sample solution: 5.0, amount of ligand APDC: more than 430 times in mole ratio, the type and concentration of acid: 0.1 M HCl, and vibration time: 7 minutes. The addition of palladium in the HCl solution could improve the reproducibility and sensitivity by a matrix modification in the absorbance measurement. This procedure was applied for the analysis of three kinds of real water samples. The detection limits equivalent to 3 times standard deviation of blank were Co 0.36 ng/mL and Ni 0.27 ng/mL and recoveries in spiked samples were 99-102% for cobalt and 100-105% for nickel.

• Korean Journal of Food Science and Technology
• /
• v.28 no.4
• /
• pp.684-688
• /
• 1996

In order to search for antithrombotic peptides from soybean paste, the inhibitory activity of water extract of soybean paste and its peptide fractions on ADP-induced aggregation of washed platelets was assayed. Soybean paste extract treated with ultrafiltration (M.W. cut off, 3,000 daltons) was found to have inhibitory activity of 90% at the dose of $96\;{\mu}g/ml$ by the method of turbidometric aggregometer. Soybean paste extract was fractionated to 19 fractions (No. B-18) by Dowex 50W X-2 ion exchange column chromatography and activity test was performed by the microplate method. All of the fractions had antithrombotic activity $(IC_{50},\;101,000\;{\mu}g/ml)$, and most fractions had higher activity than positive control, RGDS $(IC_{50}.\;205\;{\mu}g/ml)$. Especially, basic fractions No.16-18 showed higher activity than soybean paste extract $(IC_{50},\;30\;{\mu}g/ml)$. The fraction No.16 with the highest activity $(IC_{50},\;10\;{\mu}g/ml)$ was purified and analysed for amino acid composition. The results showed that histidine, arginine, and alanine were major residues in the peptide part of the fraction.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.7 no.3
• /
• pp.163-168
• /
• 1974

The present paper deals with the degradation of nucleotides in the muscle of sea mussel, Mytilus edulis, during drying. Three kinds of samples, raw, hot-air dried, and steamed-and-hot air dried were prepared and the contents of nucleotides were determined by ion exchange chromatography on columns of Dowex 1, X8. ATP and ADP were dominant in the raw muscle showed about $8{\mu}moles/g$, dry basis, respectively. The rate of degadation of ATP was very slow during drying compared with those of fish. The accumulation of ADP and AMP were observed during drying and the amount of total nucleotides (ATP+ADP+AMP) were not decreased remarkably by drying process. IMP was not detected in the all of the samples examined, however, the contents of inosine and hypoxanthine were increased during drying. In case of inosine contents, the hot-air dried sample marked an exceedingly high value equivalent to 8 times of the raw sample whereas steamed-and-hot air dried sample showed 2 times of raw samples.

• Applied Biological Chemistry
• /
• v.15 no.2
• /
• pp.93-109
• /
• 1972

Three lots of Chung-Kook-Jang were prepared by the use of 2 strains of Bacillus subtilis and Bacillus natto. For four samples taken from each lot in 12 hrs interval changes of nitrogenous compounds, insoluble protein, water soluble protein, peptides, free amino acids, amino and ammonia nitrogens during Chung-Kook-Jang fermentation, were studied together with the changes of moisture, pH, proteolytic enzyme activity. In addition the average peptide length of the peptides of a Bacillus subtilis lot was determined by the method of molecular sieving using ion exchange resin. The results were as follows: 1. The contents of moisture and total-nitrogen changed little in all samples throughout the fermentation as it would be expected. 2. In all three experimental lots the pH became higher gradually from the initial value of 6.65 to the final $7.5{\sim}7.85$ during the fermentation. Proteolytic enzyme activities, in accordance with this pH change, steadily increased up to $48{\sim}60$ hrs. of fermentation and then slightly decreased, probably affected by the high pH. The most strong proteolytic activity was observed in the experimental Chung-Kook-Jang fermentation lot using the Bacillus subtilis K-27 isolated by the author. 3. The contents of insoluble protein nitrogen in soybeans increased markedly (5%) by the cooking, after steeping 12 hrs in water. During the Chung-Kook-Jang fermentation, however, it decreased from 1/2 to 1/10 of that of the cooked soybeans. 4. The contents of water soluble protein nitrogen (5%) whereas, greatly decreased to the value of 1.0% by the cooking; but little changed further during the fermentation, 5. The total contents (0.25%) of peptides, amino, and ammonia-nitrogens, PAA-N., increased almost double by the cooking and steadily became higher as the fermentation proceeded, reaching finally up to$4{\sim}7%$ in 72 hrs fermentation. 6. The amounts of free amino acids of soybean generally decreased during the processing of cooking, even some of them like glutamic acid were destroyed completely, However in the subsequent Chung-Kook-Jang fermentation for 72 hrs., they showed from several to a few hundreds folds increases depending upon the kinds of amino acids. Valine which was contained in HCl-hydrolyzed steeped or cooked soybeans in amounts $220{\sim}267mg%$ was not detected at all as the free amino acid in all fermented samples. 7. Average peptide length (APL) of all fractions, eluted and fractionated by using the Dowex-50 ion exchange resin column, and fraction collector showed the highest value for the cooked soybean and then decreased as the fermentation proceeded. The APL value of effluent showed the highest in 12 hrs fermented sample, The value decreased thereafter by fermentation.

• Korean Journal of Agricultural Science
• /
• v.9 no.2
• /
• pp.584-590
• /
• 1982

Copolymerization of the 4-vinylpyridine with vinylacetate and divinylbenzene initiated by azobis-isobutyronitrile was carried out in DMF in presence $BaCl_2$ at $98^{\circ}C$. Ion exchange res in, poly 4-vinylpyridine-vinylsulfonic acid-divinylbenzene was prepared by sulfonation of 4-vinylpyridine-vinylacetatp-divinylbenzene with concentrated sulfuric acid. The compositions of each synthetic resin were identified by means of ir adsorption spectroscopy. Anion and cation capacities of 4-vinylpyridine-vinylsulfonic acid-divinylbenzene ion exchanger were 2.5meq/g and 4.8meq/g, respectively. Adsorption of Cd(II) and Cu(II) ions have showed larger quantity in alkalie media. A study also was made of the influence of alcohol on the distribution coefficient of Cd(II) and Cu(II) ions between the synthetic ion exchanger, and solution containing hydrochloric acid, various alcohols and water. The distribution coefficients of metal ions decrease generally as the number of branches of carbon in the molecule of butyl alcohol increase. (t-BuOH

• Applied Biological Chemistry
• /
• v.8
• /
• pp.11-20
• /
• 1967

In order to study the change of nitrogeneous compounds during the "Natto" preparation, the contents of insoluble protein, water soluble protein and amino nitrogen were determined and the pattern of peptides and amino acids was investigated by paper chromatography for the fractions resulting from molecular sieving. The results are summarized as follows: 1. Insoluble protein nitrogen which was increased to 84% by autoclaving the native soybean decreased to 44%, whereas the trichloroacetic acid soluble nitrogen increased from 8% to 45% during Natto preparation, But the soluble protein nitrogen showed a slight increase. 2. Fractionation of the peptides using Dowex-50 resins showed that they consisted mostly of lower molecular weight peptides which increased in accordance with the progress of fermentation, especially after 30-hour period. 3. Sixteen known and two unknown amino acids, and three peptides with different Rf values were identified during the Natto preparation. Their appearance showed some difference in that phenylalanine appeared after 10 hours, methionine, after 20 hours and proline, after 30 hours, respectively. The three peptides appeared at the different stage of fermentation.

• KSBB Journal
• /
• v.11 no.5
• /
• pp.606-612
• /
• 1996

A specific ammino auid in a mixture can be crystallized inside an ion exchange column when displacer concentration is high enough to concentrate the amino acid in a pure band beyond its solubility limit. Glutamic acid formpd a discrete crystal layer in a cation exchanger column by operating displacement development mode and using a high concentration of displacer NaOH. The glutamic acid crystal formed was eluded from the column with the effluent stream and collected in a fraction collector. When 1.0 M of NaOH was used as a displacer, more than 60% of the loaded glutamic acid was recovered as crystal. The continuous crystallization and dissolution of crystal occurred, resulting in apparent movement of the crystal along the column without clogging or pressure increase. NaOH was proved a better displacer than NaCl because hydroxide ions neutralized hydrogen ions released from the resin and thus reduced the number of hydrogen ion competing with sodium ion for re-adsorption. The displacement development process coupled with crystallization provided higher concentration and recovery of glutamic acrid than conventional chromatography.