• Applied Biological Chemistry
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• v.46 no.4
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• pp.380-384
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• 2003

Optimal extraction, separation and capillary rinsing conditions for capillary electrophoresis (CE) were established to discriminate the geographical origin of ligusticum root (Ligusticum wallichii) using 113 samples (domestic sample n = 62, foreign sample n = 51). Ligusticum root was extracted with 30% ethanol and separated on a uncoated fused-silica $(50\;{\mu}m{\times}27\;cm)$ capillary. Conditions for optimal analysis include: temperature, $40^{\circ}C$; voltage, 10 kV; and pressure injection time, domestic and foreign samples were 5 sec and 2 sec, respectively. The optimal separation buffer was 0.1 M phosphate buffer (pH 2.5) containing 15 mM iminodiacetic acid with 40% methanol. Under the optimal conditions established for CE, the ratio of specific peak area (peak LW-1) to other peak area (peak LW-5) was effective in discrimination geographical origin of ligusticum root. The mean accuracy for correct discrimination of geographical origin of domestic and foreign ligusticum roots were 65% and 63%, respectively.

• Analytical Science and Technology
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• v.20 no.5
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• pp.424-433
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• 2007

An analytical method for the determination of thyroid hormones in plasma samples has been studied by solid-phase extraction and high-performance liquid chromatography/diode array detector (DAD)/electrospray ionization (ESI)-mass spectrometer. Seven thyroid hormones were successfully separated by gradient elution on the reverse phase Hypersil ODS column (4.6 mm I.D., 250 mm length, particle size $5{\mu}m$) with ammonium formate buffer and acetonitrile. In addition, these compounds were confirmed by UV spectra and ESI-mass Spectra. The extraction recoveries of thyroid hormones in the plasma sample (at pH 3) were in the range of 74.5-115.7 % with solid-phase extraction by C18, followed by elution with 4 mL of methanol. The calibration curves showed good linearity with the correlation coefficients ($r^2$) varying from 0.9939 to 0.9978 and the detection limits of all analytes were obtained in the range of 20-50 ng/mL (38.1-162.8 pmol/mL). As a result, thyroxine was found in the range of 50.98-112.97 ng/mL in normal plasma samples.

• Preventive Nutrition and Food Science
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• v.7 no.4
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• pp.437-441
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• 2002

Rice proteins were extracted from brown and milled rice of five varieties: Kwanganbyeo, Daeanbyeo, Daejinbyeo, Surabyeo, Hwaseongbyeo; and their electrophoretic patterns were analyzed by SDS-PAGE. Albumin was extracted with water, globulin with 5% NaCl, prolamin with 70% ethanol, and glutelin with 0.2 M sodium borate buffer (pH 10.0) containing 0.5% SDS, 0.6% $\beta$-mercaptoethanol. The ratios of albumin : globulin : prolamin : glutelin in the brown rice were 10.8~14.1 : 12.4~16.4 : 3.6~5.3 : 68.6~72.8, and in milled rice were 4.4~5.6 : 10.6~12.0 : 3.9~5.4 : 75.7~79.8. In albumin seven major bands were observed with molecular weights ranging from 14.g~96.8 kDa, in globulin four bands with molecular weights in the range of 14.4~56.9 kDa, prolamin had only one band with a molecular weight of 14.4 kDa, and glutelin had four bands with molecular weights of 14.4 ~ 57.4 kDa. There were no differences in electrophoretic patterns between rice varieties or between brown and milled rice.

• Microbiology and Biotechnology Letters
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• v.51 no.1
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• pp.99-108
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• 2023

Several challenges arise in DNA extraction and gene amplification for airborne fungal metagenome analysis from a particulate matter (PM) samples. In this study, various conditions were tested to optimize the DNA extraction method from PM samples and polymerase chain reaction (PCR) conditions with primer set and annealing temperature. As a result of comparative evaluation of DNA extraction under various conditions, chemical cell lysis using buffer and proteinase K for 20 minutes and bead beating treatment were followed by using a commercial DNA extraction kit to efficiently extract DNA from the PM filter samples. To optimize the PCR conditions, PCR was performed using 10 primer sets for amplifying the ITS2 gene region. The concentration of the PCR amplicon was relatively high when the annealing temperature was 58℃ with the ITS3tagmix3/ITS4 primer set. Even under these conditions, when the concentration of the PCR product was low, nested PCR was performed using the primary PCR amplicon as the template DNA to amplify the ITS2 gene at a satisfactory concentration. Using the methods optimized in this study, DNA extraction and PCR were performed on 15 filter samples that collected PM2.5 in Seoul, and the ITS2 gene was successfully amplified in all samples. The optimized methods can be used for research on analyzing and interpreting the fungal metagenome of atmospheric PM samples.

• Parasites, Hosts and Diseases
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• v.56 no.1
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• pp.25-32
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• 2018

Molecular techniques have been introduced for malaria diagnosis because they offer greater sensitivity and specificity than microscopic examinations. Therefore, DNA isolation methods have been developed for easy preparation and cost effectiveness. The present study described a simple protocol for Plasmodium DNA isolation from EDTA-whole blood. This study demonstrated that after heating infected blood samples with Tris-EDTA buffer and proteinase K solution, without isolation and purification steps, the supernatant can be used as a DNA template for amplification by PCR. The sensitivity of the extracted DNA of Plasmodium falciparum and Plasmodium vivax was separately analyzed by both PCR and semi-nested PCR (Sn-PCR). The results revealed that for PCR the limit of detection was $40parasites/{\mu}l$ for P. falciparum and $35.2parasites/{\mu}l$ for P. vivax, whereas for Sn-PCR the limit of detection was $1.6parasites/{\mu}l$ for P. falciparum and $1.4parasites/{\mu}l$ for P. vivax. This new method was then verified by DNA extraction of whole blood from 11 asymptomatic Myanmar migrant workers and analyzed by Sn-PCR. The results revealed that DNA can be extracted from all samples, and there were 2 positive samples for Plasmodium (P. falciparum and P. vivax). Therefore, the protocol can be an alternative method for DNA extraction in laboratories with limited resources and a lack of trained technicians for malaria diagnosis. In addition, this protocol can be applied for subclinical cases, and this will be helpful for epidemiology and control.

• YAKHAK HOEJI
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• v.31 no.2
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• pp.45-51
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• 1987

Methyl orange(MO) and cetyltrimethylammonium bromide(CTA) were mixed and added to 10ml of the buffer solution and then the solution was shaken for 5 minutes. The maximum absorption wavelength of the reaction product was 419nm. Dichloromethane was the best extracting solvent among the several organic solvents and the most suitable pH range was 2~8. When the CTA-MO calibration curve was made in the best experimental condition, the Lambert-Beer's law was obeyed in the range of CTA concentration of $1.5\times$$10^{-5}$~1.0$\times$$10^{-4}M$ by UV spectrophotometer. This method was possible to determine quaternary ammonium salts in the pharmaceutical preparations.

• Bulletin of the Korean Chemical Society
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• v.23 no.5
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• pp.693-698
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• 2002

A method has been described for the determination of Cu(Ⅱ), Pb(Ⅱ), Ni(Ⅱ), Cd(Ⅱ), Mn(Ⅱ) and Fe(Ⅲ) by flame atomic absorption spectrometry (FAAS) after preconcentration on Amberlite XAD-16 resin, using hexamethyleneammonium-hexamethylenedithiocarbamate (HMA-HMDTC) as a chelating agent, and NH3/NH4Cl buffer solution (pH 9). Influences of various analytical parameters such as pH, concentration of nitric acid, amount of analytes, diverse ions and sample volume were investigated. The relative standard deviation (RSD) and the detection limit (LOD) were found in the range of 0.8-2.9% and 0.006-0.277 ㎍/mL,respectively. Recoveries obtained by the column method were quantitative ( >95%) at optimum conditions.The method was applied to the determination of some metal ions in seawater and wastewater samples. A high preconcentration factor (about 150 for seawater and 75 for wastewater samples) and simplicity are the main advantages of this suggested method.

• YAKHAK HOEJI
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• v.30 no.6
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• pp.343-347
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• 1986

Korean ginseng was purified to obtain radioprotective protein fractions by buffer extraction, ammonium sulfate fractionation, CM-cellulose column chromatography, heat inactivation and Sephadex G-75 column chromatography. The final three fractions, GI, GII and GIII were subjected to Disc-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE. The molecular weights(M.W.) of native and denatured proteins were estimated by using regression line equations obtained from the mobilities of standard proteins. As the results, in Disc-PAGE, the GI fraction showed two protein bands with M.W. of above 213, 000 and 55, 000, GII showed one band with M.W. of 44, 000 and GIII, also one band with M.W. of 19, 000. In SDS-PAGE, GI fraction gave four subunit bands with M.W. of above 114, 000, 27, 000, 24, 000 and 19, 000, GII gave two bands with M.W. of 46, 000 and 22, 000, and GIII, one band of 19, 000.

• Archives of Pharmacal Research
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• v.21 no.2
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• pp.217-222
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• 1998

A stereospecific HPLC method has been developed for the resolution of the enantiomers of salbutamol in human urine. After solid-phase extraction and derivatization with 2,3,4,6-tetra-O-acetyl-$\beta$-D-glucopyranosyl isothiocyanate, the diastereomeric derivatives were resolved (Rs=1.83) on $5{\mu}m$ octadecylsilan column using 35% acetonitrile in 0.05M ammonium acetate buffer (pH=6) as a mobile phase with electrochemical detection. The diastereomeric derivatives were formed within 30 min. The detection limit of each enantiomer was 20 ng/ml (S/N=3).

• The Journal of the Korean Society for Microbiology
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• v.21 no.2
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• pp.227-231
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• 1986

The hemagglutination activities of Hantaan virus and Seoul virus were demonstrated. The hemagglutinins were prepared by sucrose-acetone extraction method from suckling mouse brains infectecd with Hantaan and Seoul viruses. Hemagglutination of goose erythrocytes by these viral hemagglutinins was pH dependent in phosphate buffer system. Hantaan and Seoul viruses were distinguished by pH range of hemagglutination. 76/118 and 79/90 strains of Hantaan virus showed hemagglutination at the range of pH 5.75-6.4 and the optimal pH was 5.75 with the titer of 1:512 in 76/118 and 1:256 in 79/90. In contrast, KSNUSD 84/34 strain of Seoul virus revealed hemagglutination at the range of pH 6.2-6.4 and the optimal pH was 6.4 with the titer of 1 : 64.