• Applied Biological Chemistry
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• v.15 no.2
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• pp.111-142
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• 1972

An experimental Chung-Kook-Jang was prepared using the strain Bacillus subtilis sp. isolated by the author Samples were taken in 12 hrs interval during the fermentation and the oligopeptides were separated by the method of molecular sieving using the ion exchange resin column of Dowex-50. Only the X-16 fraction of oligopeptides was taken and the components of oligopeptides were developed in two dimensional thin layer chromatograms. The each peptide spot was eluted and each peptide was isolated. The pattern and kinds of amino acids, and N and C-terminal amino acids were studied. Fourteen different oligopeptides could be detected by the two dimensional thin layer chromatography, all of which were consisted of $4{\sim}9$ kinds of amino acids. No dipeptides and no tripeptides could be found. The N and C-terminal amino acids and the residual component amino acids of all these 14 peptides could be summarized as the follows. [P]-I. Pro (Cys Ala Asp Trp Ile Val) Glu [P]-II. Val (His Arg Glu Thr Ala Met) Asp [P]-III. Glu (Cys Lys Asp Thr Met) Ala [P]-IV. Glu(His Ser Ala) Met) [P]-V. Ile (Cys Asp Arg Gly Pro T.p Phe) His [P]-VI. Gly(Asp ser) Lys [P]-VII. Thr(Pro Tyr Phe) Asp [P]-VIII. Phe(Tyr Leu Ile) Val [P]-IX. Trp (Phelle) Thr [P]-X. Ile (Arg Leu) Phe [P]-XI. Asp (Lys His Ser Gly Glu Pro) Ala [P]-XII. Glu (Cys Asp Gly) Ser [P]-XIII. Ala (Arg Tyr) Glu [P]-XIV. Met (Glu Ala) His It appears that the protease of the Bacillus subtilis K-27 syrain has rather wider range of specificity than proteases of Aspergoillus soya, pepsin, chymotrypsin, and trypsin.

• Korean Journal of Environmental Agriculture
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• v.31 no.2
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• pp.157-163
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• 2012

BACKGROUND: For the safety of imported agricultural products, the study was conducted to develop the analytical method of unregistered pesticides in domestic. The analytical method of 6 pesticides, chlorthal-dimethyl, clomeprop, diflufenican, hexachlorobenzene, picolinafen, and propyzamide, for a fast multi-residue analysis were established for two different type crops, orange and brown rice by GC-ECD and confirmed by mass spectrometry. METHODS AND RESULTS: The analytical method was evaluated to limit of quantification, linearity and recoveries. The crop samples were extracted with acetonitrile and performed cleanup by liquid-liquid partition and Florisil SPE to remove co-extracted matrix. The extracted samples were analyzed by GC-ECD with good sensitivity and selectivity of the method. The limits of quantification (LOQ) range of the method with S/N ratio of 10 was 0.02~0.05 mg/kg for orange and brown rice. The linearity for targeted pesticides were $R^2$ >0.999 at the levels ranged from 0.05 to 10.0 mg/kg. The average recoveries ranged from 74.4% to 110.3% with the percentage of coefficient variation in the range 0.2~8.8% at two different spiking levels (0.02 mg/kg and 0.2 mg/kg, 0.05 mg/kg and 0.5 mg/kg) in brown rice. And the average recoveries ranged from 77.8% to 118.4% with the percentage of coefficient variation in the range 0.2~6.6% at two different spiking levels (0.02 mg/kg and 0.2 mg/kg, 0.05 mg/kg and 0.5 mg/kg) in orange. Final determination was by gas chromatography/mass spectrometry/selected ion monitoring (GC/MS/SIM) to identify the targeted pesticides. CONCLUSION: As a result, this developed analytical method can be used as an official method for imported agricultural products.

• Analytical Science and Technology
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• v.23 no.2
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• pp.187-195
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• 2010

To assess health risk for the intake among residents after the Hebei Spirit oil spill, 16 Polycyclic Aromatic Hydrocarbons (PAHs) in seafood samples from oil contaminated bay were determined by Gas Chromatography-Mass Spectrometry (GC-MSD) and samples were personally collected and purchased by residents. Samples were hydrolyzed with KOH and extracted with methylene chloride. The extracted solution were cleaned up using silica/florisil column and 16 PAHs were eluted by methylene chloride : n-hexane (1:9) mixture and determined by GC-MSD in Selected Ion Monitoring (SIM) mode. The mean recoveries for 16 PAHs ranged from 79% to 85%. The 16 PAHs levels in 126 samples ranged from 0.17 to $6.04\;{\mu}g$/kg and the TEQBaP (Toxic EQuivalents) levels in 126 samples were calculated using benzo(a)pyrene toxic equivalency factor for individual 16 PAHs and ranged from 0 to $0.91\;{\mu}gTEQ$/kg. The average Benzo(a)pyrene dietary exposure of residents was $5.5{\times}10^{-8}\;mg/kg$ bw/day and the average PAHs chronic dietary exposure was $1.3{\times}10^{-5}\;mg$ TEQ/kg bw/day. The margin of exposure (MOE) and the excess cancer risk and were $1.8{\times}10^6$ and $9.8{\times}10^{-8}$, respectively. Therefore, the assessment result was considered as low concern for health risk.

• Journal of Food Hygiene and Safety
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• v.25 no.4
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• pp.388-394
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• 2010

Metabolomics aims at the comprehensive, qualitative and quantitative analysis of wide arrays of endogenous metabolites in biological samples. It has shown particular promise in the area of toxicology and drug development, functional genomics, system biology and clinical diagnosis. In this study, analytical technique of MS instrument with high resolution mass measurement, such as time-of-flight (TOF) was validated for the purpose of investigation of amino acids, sugars and fatty acids. Rat urine and serum samples were extracted by selected each solvent (50% acetonitrile, 100% acetonitrile, acetone, methanol, water, ether) extraction method. We determined the optimized liquid chromatography/time-of-flight mass spectrometry (LC/TOFMS) system and selected appropriated columns, mobile phases, fragment energy and collision energy, which could search 17 metabolites. The spectral data collected from LC/TOFMS were tested by ANOVA. Obtained with the use of LC/TOFMS technique, our results indicated that (1) MS and MS/MS parameters were optimized and most abundant product ion of each metabolite were selected to be monitorized; (2) with design of experiment analysis, methanol yielded the optimal extraction efficiency. Therefore, the results of this study are expected to be useful in the endogenous metabolite fields according to validated SOP for endogenous amino acids, sugars and fatty acids.

• Applied Biological Chemistry
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• v.39 no.6
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• pp.494-500
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• 1996

Uptake of hen metal ions by water dropwort (Oenanthe stolonifera DC.) and its cadmium-binding protein were studied to probe for good method to remove heavy metal contaminants from environments. The plant was cultured in the culture medium (pH 7.0) containing the various concentrations of $Cd^{2+}$, $Cr^{3+}$ or $Pb^{2+}$, for 3 and 7 days. The residual heavy metals deposited in roots linearly increased as the metal ions concentration increased up to 17 ppm for $Cd^{2+}$, 20 ppm for $Cr^{3+}$ and 50 ppm for $Pb^{2+}$. Above these concentrations, the plant growth was inhibited and the uptake rates of the metal ions decreased. The heavy metals absorbed by the plant were mostly deposited in roots. In particular, the residual concentration of lead in roots was about four times higher than those of cadmium and chromium. When cultured in the medium containing 20 ppm of each metal ion, 80% of cadmium, 90% of cromium and 96% of lead were deposited in roots out of the total residual metal ions in the plant. These values correspond to 6.1 mg of cadmium, 5.2 mg of chromium and 23.6 mg of lead per one gram of roots tissue on a dry weight basis. A cadmium-binding protein was partially purified by extraction, gel filtration and DEAE-Cellulose chromatography from water dropworts that was grown in the medium containing 20 ppm $Cd^{2+}$. The purified protein was a single band on SDS- and non-denaturing- polyacrylamide gel electrophoresis. Its molecular mass was estimated to be ca. 5,000 dalton by gel filteration. Analysis of amino acid composition of the protein indicated that it had a typical amino acid composition of heavy metal-binding protein in that it contained 27% of acidic amino acids and 9.9% of cysteine. However, it is likely that the protein is a new plant metal-binding protein, since its amino acid composition is somewhat different from those of phytochelatins that have been known so far.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.9
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• pp.1256-1264
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• 2011

Infant formula is one of the main nutritional sources for infants. In this study, the fatty acid compositions of Korean (domestic, n=8) and foreign infant formulas (n=3) were analyzed. Crude fats were extracted using the Folch method, and fatty acid compositions (total and positional) were analyzed by gas chromatography. In the fatty acid compositions of infant formulas, oleic (19.88~44.64%), palmitic (7.59~21.65%) and linoleic acids (16.72~25.87%) were the major fatty acids. Domestic infant formula products contained 35.94~56.55% total saturated fatty acid content, whereas that of foreign infant formula ranged from 34.40~42.88%. The content of monounsaturated fatty acids ranged from 20.23~44.99% in domestic products and 34.11~45.07% in foreign products. In addition, 0.17~2.57% arachidonic acid/docosahexaenoic acid and 10~13% linoleic acid/linolenic acid were detected in the analyzed products (domestic and foreign products). A small amount of trans fatty acids (0.25~1.69%) were found. In sn-2 position analysis, palmitic acids (1.84~38.74%) were detected in the analyzed formulas. Further, typical triacylglycerols in human milk, including 1,3-di-monounsaturated-2-saturated triacylglycerol, were not detected in the analyzed formulas.

• The Korean Journal of Pesticide Science
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• v.19 no.1
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• pp.5-13
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• 2015

A chromatographic method for the determination of imazapyr, a non-selective herbicide, in agricultural commodities was developed to use safety control of pesticide residue on crops, and was fully validated as an official method for residue analysis. Agricultural commodities, mandarin (fruit), hulled rice (cereal grains), pepper (vegetables), potato (potatoes) and soybean (beans) were extracted with methanol and partitioned with dichloromethane to remove the interference obtained from sample extracts, adjusting pH to 2.5 by 4N hydrochloric acid. Finally, they were analyzed by high performance liquid chromatography coupled to UV detector (HPLC-UVD). The developed method had the linearity in the range of test concentrations with coefficients of determination ($r^2$) more than 0.99. Recovery studies were carried out at three concentration levels (LOQ, 10LOQ, and 50LOQ) performing five replicates at each level. Recoveries were ranged between 72.1 to 108.0%, with relative standard deviations less than 10%. A consistent recovery was determined according to the CODEX guidelines (CAC/GL40, 2003). Finally, LC/MS with selected ion monitoring was also applied to confirm the suspected residues of imazapyr in agricultural samples. This developed method for determination of imazapyr residues in agricultural commodities. can be used as an official method.

• Applied Biological Chemistry
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• v.5
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• pp.23-31
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• 1964

Six species of marine shell-fishes were subjects in this study. The content of RNA and DNA in mid-intestinal glands (liver) was determined and RNA was also extracted from above materials by phenol method and their nucleotide compositions were analysed by ion exchange column chromatography. C; citidylic acid, A; adenylic acid, G; guanylic acid, U; uridylic acid, Pu; purine nucleotide, Py; pyrimidine nucleotide 1) Their RNA and DNA content was summarized in the next table.(mg/100mg of lipid free powder) Materials RNA DNA RNA/DNA Meretrix meretrix Susoria (Gmelin) 6.82 0.82 6.5 Anadara(scapharea) inflata(Reeve) 4.62 0.70 6.6 Ostrea(crassostrea) gigas Thunberg 8.74 1.03 8.5 Turbo cornutus Solander 2.16 0.60 3.6 Haliotis gigantea Gmelin 7.02 0.22 36.0 2) Their RNA nucleatide compositions was summaries in following tabe. Materials/RNA nucleotide G+C/A+U G+U/A+C Pu/Py Meretrix meretrix Susoria(Gmelin) 1.12 1.36 1.08 Venerupis philippinarum (Adoms et Reeve) 1.15 1.31 1.13 Anadara(scapharea) inflata(Reeve) 0.97 1.26 1.02 Ostrea (crassostrea) gigas Thunberg 1.51 1.34 1.31 Turbo cornutus Solander 1.37 1.29 1.03 Haliotis gigantea Gmelin 1.33 1.46 1.22 In six species of marine fishes examined, Pu/Py and G+C/A+U ratios of RNA vary in respective wide ranges of 1.02-1.31, 0.97-1.51, while G+U/A+C ratio in the range of 1.26-1.46, not far from 1.3. This G+U/A+C ratio seems to be specific in this species.

• Journal of Food Hygiene and Safety
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• v.29 no.2
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• pp.131-140
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• 2014

The objective of this study was to develop a simultaneous method of 8 penicillin antibiotics including amoxicillin, ampicillin, cloxacillin, dicloxacillin, nafcillin, oxacillin, penicillin G and penicillin V in meat using LC-MS/MS. The procedure involves solid phase extraction with HLB cartridge and subsequent analysis by LC-MS/MS. To optimize MS analytical condition of 8 compounds, each parameter was established by multiple reaction monitoring in positive ion mode. The chromatographic separation was achieved on a $C_{18}$ column with a mobile phase of 0.05% formic acid and 0.05% formic acid in acetonitrile at a flow rate of 0.2 mL/min for 20 min with a gradient elution. The developed method was validated for specificity, linearity, accuracy and precision in beef, pork and chicken. The recoveries were 71.0~106%, and relative standard deviations (RSD) were 4.0~11.2%. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.003~0.008 mg/kg and 0.01~0.03 mg/kg, respectively, that are below maximum residue limit (MRL) of the penicillins. This study also performed survey of residual penicillin antibiotics for 193 samples of beef, pork and chicken collected from 9 cities in Korea. Penicillins were not found in all the samples except a sample of pork which contained cloxacillin (concentration of 0.08 mg/kg) below the MRL (0.3 mg/kg).

• Korean Journal of Food Science and Technology
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• v.32 no.5
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• pp.1191-1197
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• 2000

In the preliminary study, we investigated the anti-complementary activities of 62 extracts from Korean edible seaweeds. Of those, Pachymeniopsis elliptica showed the highest anti-complementary activity. Therefore, it was purified as follows; i) PE-1 by ethanol precipitation, ii) PE-1-C by ultrafiltration, iii) PE-1-CIV by DEAE-Toyopearl 650C, and iv) PE-1-CIV-ii by Sepharose CL-6B. The purified compound, PE-1-CIV-ii, was the complexed homogeneous polysaccharide (molecular mass: 780 kDa) with 82.9% of anti-complementary activity. Also, it contained a significant amount of sulfate group (30.5%), which indicated it as a sulfated algal polysaccharide. Its structural monosaccharides were galactose (44.3%), 3,6-anhydrogalactose (34.0%), glucose (8.2%), fucose (5.4%), xylose (5.2%) and rhamnose (2.9%). After the treatment of periodate on a sample, a significant decrease in anti-complementary activity was found, which was a characteristic of bioactive polysaccharides. And-tumor activity of PE-1-A, B and C was tested in the sarcoma-180 solid tumor model. The PE-1-C with the largest molecular mass (more than 300 kDa) showed 81% of inhibition on the solid tumors, suggesting that the anti-complementary activity was, at least in part, related to anti-tumor activity. Based upon these results, the purified polysacchardes could be an immunopotentiator in vivo.