• Journal of Pharmaceutical Investigation
• /
• v.35 no.5
• /
• pp.383-388
• /
• 2005

A simple and specific method for determination of methyltestosterone (MT) has been established by a gas chromatography/mass selective detector and applied in plasma of healthy male volunteers received a single oral dose of 50 mg MT $(Testo^{TM}\;tablets,\;25\;mg)$ for bioavailability test. This method involves using liquid-liquid extraction of the sample with diethyl ether and derivatization with MSTFA. MT showed good resolution in this condition. The detection limit of quantitation was 5 ng/ml. A good linearity (r>0.996) was obtained at the range of 5-250 ng/ml of MT. Intra-day precision and accuracy were 2.76-12.56% and 0.39-8.01 %, and inter-day precision and accuracy were 2.29-17.69% and 0.42-7.99%, respectively. The established method was applied on bioavailability test of MT in human volunteers. The value of $AUC_{0\;to\;last}$ to last was $264.5{\pm}123.9\;ng{\cdot}hr/ml$ and that of $AUC_{0\;to\;inf}$ was determined to be $275.2{\pm}126.5\;ng{\cdot}hr/ml$. The values of $C_{max}$ and $T_{max}$ were $95.9{\pm}67.1\;ng/ml$ and $1.13{\pm}0.9\;hr$, respectively. The mean elimination half-life $(t_{1/2})$ was $4.4{\pm}0.9\;hr$. This analytical method is suitable and useful for the pharmacokinetics and bioequivalence studies of MT.

• Applied Microscopy
• /
• v.3 no.1
• /
• pp.45-54
• /
• 1973

The present study is to determine the toxicity of the fungi isolated from foodstuffs by observing the ultrastructural changes in the mouse liver cells. The results as follows: 1. The toxin-producing fungi were screened by the methods of toxin-screening test(cyto-toxicity test against to HeLa cells and thin layer chromatography). 2. All of the experimental animals treated with isolated fungi were observed the focal necrosis and inflammatory infiltration of liver parenchymal cells. 3. It showed the cytoplasmic changes, such as dilatation of rough endoplasmic reticulum (RER), swelling of mitochondria (mi). increased number of lipid droplet (li) and glycogen (gl), detachment of ribosomes (ri) by observing the electron microscopy. 4. Nuclear and nucleolar alteration were also noted the segregation of nucleolar element and irregularity of nuclear envelopes. 5. As a mass screening, the cytotoxicity test using HeLa cells and thin layer chromatography are feasible methods to detection of the mycotoxin producing fungi from various sources.

• Journal of Pharmacopuncture
• /
• v.18 no.1
• /
• pp.44-55
• /
• 2015

Objectives: Scolopendra subspinipes mutilans (S. subspinipes mutilans) is known as a traditional medicine and includes various amino acids, peptides and proteins. The amino acids in the pharmacopuncture extracted from S. subspinipes mutilans by using derivatization methods were analyzed quantitatively and qualitatively by using high performance liquid chromatography (HPLC) over a 12 month period to confirm its stability. Methods: Amino acids of pharmacopuncture extracted from S. subspinipes mutilans were derived by using O-phthaldialdehyde (OPA) & 9-fluorenyl methoxy carbonyl chloride (FMOC) reagent and were analyzed using HPLC. The amino acids were detected by using a diode array detector (DAD) and a fluorescence detector (FLD) to compare a mixed amino acid standard (STD) to the pharmacopuncture from centipedes. The stability tests on the pharmacopuncture from centipedes were done using HPLC for three conditions: a room temperature test chamber, an acceleration test chamber, and a cold test chamber. Results: The pharmacopuncture from centipedes was prepared by using the method of the Korean Pharmacopuncture Institute (KPI) and through quantitative analyses was shown to contain 9 amino acids of the 16 amino acids in the mixed amino acid STD. The amounts of the amino acids in the pharmacopuncture from centipedes were 34.37 ppm of aspartate, 123.72 ppm of arginine, 170.63 ppm of alanine, 59.55 ppm of leucine and 57 ppm of lysine. The relative standard deviation (RSD %) results for the pharmacopuncture from centipedes had a maximum value of 14.95% and minimum value of 1.795% on the room temperature test chamber, the acceleration test chamber and the cold test chamber stability tests. Conclusion: Stability tests on and quantitative and qualitative analyses of the amino acids in the pharmacopuncture extracted from centipedes by using derivatization methods were performed by using HPLC. Through research, we hope to determine the relationship between time and the concentrations of the amino acids in the pharmacopuncture extracted from centipedes.

• KSBB Journal
• /
• v.25 no.3
• /
• pp.205-214
• /
• 2010

Many successive liquid-liquid extractions occur enabling purification of the crude material to occur. In high performance counter-current chromatography (HPCCC), crude material is partitioned between two immiscible layers of solvent phases. The stationary phase (SP) is retained by hydrodynamic force field effect and the mobile phase (MP) is pumped through the column. Purification occurs because of the different solubility of the components in the liquid mobile and stationary phases. There are many key benefits of liquid stationary phases such as high mass and volume injection loadings, total sample recovery, and easy scale-up. Many researchers showed that predictable scale-up from simple test is feasible with knowledge of the stationary phase retention for the planned process scale run. In this review we review the recent advances in HPCCC research and also describe the key applications such as natural products and synthetics (small or large molecules).

• Korean Journal Plant Pathology
• /
• v.14 no.3
• /
• pp.191-202
• /
• 1998

About 300 actinomycetes were isolated from two forest and one sea-shore soil and tested for inhibitory effects on mycelial growth of six plant pathogenic fungi Magnaporthe grisea, Alternaria mali, Colletotrichum gloeosporioides, Phytophthora capsici, Fusarium oxysporum f. sp. cucumerinum, and Rhizoctonia solani. Among 300 actinomycetes tested, only 16 actinomycetes showed the antifungal activity against the test fungi. Isolate NH 50 was selected for production and purification of antifungal antibiotic substances. Actinomycete isolate NH 50 displayed the broad antifungal spectra against 11 plant pathogenic fungi. To identify actinomycete isolate NH 50, cultural characteristics on various agar media, diaminopimelic acid type, and morphological characteristics by scanning electron microscopy were examined. As a result, actinomycete isolate NH 50 was classified as a rare actinomycete that had LL-DAP type and did not produce spores. After incubation of isolate NH 50 in yeast extract-malt extract-dextrose broth, antifungal compound NH-B1 that inhibited mycelial growth of some plant pathogenic fungi was purified from the methanol eluates of XAD-16 resins by a series of purification procedures, i.e., silica gel flash chromatography, C18 flash chromatography, Sephadex LH-20 column chromatography, silica gel medium pressure liquid chromatography (MPLC), C18 MPLC, and high pressure liquid chromatography (HPLC). UV spectrum and 1HNMR spectrum of antifungal compound NH-B1 dissolved in methanol were examined. The antibiotic NH-B1 showed the major peaks at 230 and 271.2nm. Based on the data of 1H-NMR spectrum, NH-B1 was confirmed to be an extremely complex polymer of sugars called polysaccharides. The antibiotic NH-B1 showed strong antifungal activity against Alternaria solani and Cercospora kikuchi, but weak activity against M. grisea.

• Journal of the Korean Wood Science and Technology
• /
• v.29 no.4
• /
• pp.79-88
• /
• 2001

Lectin was isolated from shiitake mushroom (Lentinula edodes) with 0.15 M NaCl solution, and purified by the following procedures : precipitation by ammonium sulfate, anion exchange column chromatography on DEAE Sephadex A-50 and hydroxyapatite column chromatography. The fresh pileus part of the mushroom contained more than two times of lectin compared to the stipe part, and lectins and its activity were reduced by heating. The extraction yield of crude lectin was 46.03%, 28% yield after purification on on DEAE Sephadex A-50 column chromatography. Some amino acids, aspartic acid, serine, alanine and histidine, were increased by purification process. Relatively low molecular weight parts of lectin had the agglutinating activity for rabbit blood, and its molecular weight was about 23 kDa The molecular weights of purified lectins, LA-a and LB-b, by the hydroxyapatite column chromatography were 24 kDa and 23 kDa, respectively.

• Journal of Food Hygiene and Safety
• /
• v.12 no.1
• /
• pp.71-77
• /
• 1997

Sulfonamides, a therapeutically important group of antimicrobial drugs, are widely used to treat and prevent the acute systemic and skin infections in dairy cattle. They also pose an economic hazard through inhibition of growth of dairy starter cultures. This study was carried out to compare four screening methods for detection of sulfamethazine in milk and determine the positive milk sample by HPLC method. Sulfamethazine was used to spike at five levels of sulfamethazine. The Lac-Tek test and CharmII test were also consistent better than TTCII test and Delvo SP test in sulfamethazine detection. Analysis probabilities of obtaining a positive response with TTCII test and Delvo SP test assay at 50 ppb sulfamethazine level in milk samples were only 14%, 42% each. Whereas using the Lac-Tek test and CharmII test would have resulted in 100% identification of the five levels. Determination of sufamethazine using the HPLC method in the spiked milk were 10.64, 19.30, 30.76,38.83 and 50.23 ppb, respectively.

• Applied Biological Chemistry
• /
• v.51 no.3
• /
• pp.153-158
• /
• 2008

An actinomycetes F-97 producing tyrosinase inhibitor was isolated from soil samples. Isolation and purification of tyrosinase inhibitor produced by F-97 was performed as follows: IRC-120 ($NH_4^+$ type) column chromatography, silica gel column chromatography, $C_{18}$ column chromatography and Sephadex LH-20 column chromatography were used successively after the centrifuged supernatant was adjusted to pH 4.0. To identify the purity of the inhibitor, octadecylsilyl(ODS) HPLC was carried out with 5% methanol as a mobile phase. Finally, the purification yield of a tyrosinase inhibitor was 5.24%. The inhibitor was very soluble in water, methanol and ethanol but insoluble in acetone, butanol, ethylacetate and chloroform. The ${\lambda}_{max}$ value of this inhibitor in water was 194nm under UV light. The biochemical test of the inhibitor was positive in Molish, Benedict, cone. $H_2SO_4$, and $KMnO_4$ tests but negative in iodine, ninhydrin, Million, Sakaguchi, xanthoproteic and Emerson tests. The tyrosinase inhibitor was stable against heat treatment of $100^{\circ}C$ for 50 minutes and pH $4{\sim}9$. The $IC_{50}$ value of this inhibitor was $19.2{\mu}g/ml$ for mushroom tyrosinase. In $1,000{\mu}g/ml$ inhibitor concentration, inhibition zone was 27 mm for Streptomyces bikiniensis NRRL B-1049. The inhibition of F-97 against mushroom tyrosinase was competitive with tyrosine.

• Korean Journal of Pharmacognosy
• /
• v.41 no.2
• /
• pp.136-140
• /
• 2010

This study was carried out to utilize non-toxic solvents instead of harmful solvents for the TLC(Thin Layer Chromatography) identification test of herbal medicines. It is recommended not to use harmful solvent such as chloroform at a viewpoint of clean analysis. In this study, we revised the identification test of 10 items in the Korea Pharmacopoeia(KP) and the Korean Herbal Pharmacopoeia(KHP) such as Cornus officinalis S. containing the harmful solvents on the developing solvent and established identification test that is utilized non-toxic solvents by HPTLC(High Performance Thin Layer Chromatography).

• Korean Journal of Microbiology
• /
• v.25 no.1
• /
• pp.73-79
• /
• 1987

A restriction endonuclease, Kpn I has been isolated from Klebsiella pneumonia. Cells were broken by sonication. After ultracentrifugation the supernatant containing Kpn I activity was further purified by Sepharose-6B gel filtration, DEAD-Cellulose, Heparin-Agarose, and Aminohexyl-Agarose column chromatography. Final enzyme preparation was essentially free of contamination exonuclease and phosphatase, as judged by ligation-recut test. Total activity of the enzyme recovered from 10 grams of cells was $4.7\times 10^5$ units.