• 약학회지
• /
• 제54권4호
• /
• pp.295-299
• /
• 2010

The aim of this study was to develop and validate for determination of alibendol in human plasma by HPLC method. After precipitation of 500 ${\mu}l$ plasma samples by 50% methanol 50 ${\mu}l$ and 60% perchloric acid 30 ${\mu}l$ and the supernatant 50 ${\mu}l$ was injected into HPLC. The assay was performed isocratically using 10 mM potassium phosphate (pH 3.0) and acetonitrile (80 : 20, v/v) as mobile phase. The $C_{18}$ column (particle size $3.5{\mu}m$, $4.6{\times}50$ mm, Zorbax Eclipse) was used as a solid phase. The mobile phase was delivered at a flow-rate of 1.7 ml/min, detection was by ultraviolet absorption at 232 nm and concentrations were calculated on the basis of peak areas. In these conditions, alibendol can be separated from ethylparaben, the internal standard, and endogenous substances. The retention times of alibendol and ethylparaben were just about 2.6 and 3.5 minutes, respectively. This rapid HPLC method was validated by examining the precision and accuracy for inter- and intra-day analysis. The standard curve was linear ($R^2$=1.0000) over the concentration range of 0.05~20 ${\mu}g$/ml. The inter-day relative standard deviation (R.S.D.) and accuracy were 0.2~12.2% and 94.4~101.2% (82.7% at the lower limit of quatitation). The intra-day R.S.D. and accuracy were 0.1~11.8% and 98.8~102.5%, respectively. The method was successfully applied to the determination of alibendol in plasma for a pharmacokinetic study.

• Journal of Microbiology and Biotechnology
• /
• 제28권10호
• /
• pp.1749-1759
• /
• 2018

Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

• Archives of Pharmacal Research
• /
• 제26권12호
• /
• pp.1024-1028
• /
• 2003

A prolyl endopeptidase inhibitor was isolated from the ethyl acetate soluble fraction of Phyllanthus ussurensis. The active compound was identified as an ellagitannin, corilagin. It was shown to non-competitively inhibit prolyl endopeptidase (PEP) with the $IC_{50}$ value of $1.17 \times $10^{-6}\mu$M. The Ki value was $6.70 \times 10^{-7}$ M. Corilagin was less inhibitory to other serine proteases such as chymotrypsin, trypsin, and elastase, indicating that it was relatively a specific inhibitor of PEP. Corilagin also effectively inhibited reactive oxygen species such as hydroxide and superoxide anion radical, hydrogen peroxide, and DPPH. Especially, corilagin showed potent scavel1ging activity on the superoxide anion radical in the ESR method ($IC_{50} =3.79 \times 10^{-6}$M) as well as xanthine oxidase system.

• 약학회지
• /
• 제55권2호
• /
• pp.145-153
• /
• 2011

A simultaneous detection and quantification method for determining the Phenylalkylamine derivatives, such as methamphetamine (MA), amphetamine (AM), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), ketamine (KT), norketamine (NKT), phentermine (PT), fenfluramine (FFA) and phenmetrazine (PM), in oral fluid was developed and validated according to international guidelines. The validated method was applied to actual oral fluid samples collected from drug abuse suspects. The recovery of phenylalkylamines from oral fluid collection devices was also assessed. Oral fluid specimens from 20 drug abuse suspects submitted by the police were collected using Salivette$^{TM}$, Quantisal$^{TM}$ or direct expectoration. The samples were screened using a biochip array analyzer. For confirmation, the samples were analyzed by GC-MS in selected-ion monitoring (SIM) mode after extraction using automated SPE with a mixed-mode cation exchange cartridge and derivatization with trifluoroacetic anhydride (TFAA). The results from the immunoassay were consistent with those from GC-MS. All the oral fluid samples gave positive results for MA, AM, PT and/or PM. The detection of phenylalkylamines in oral fluid can provide a better indication of recent use than urine or hair. Therefore, the oral fluid specimen was useful for demonstrating phenylalkylamines abuse in the driving under the influence of drug (DUID) as an alternative specimen for urine.

• 약학회지
• /
• 제57권6호
• /
• pp.420-425
• /
• 2013

Phentermine (PT) and phenmetrazine (PM) have been widely used as anti-obesity drugs. These drugs should be used with caution due to its close relation to amphetamine in its structure and toxicity. PT and PM, amphetamine-type anorectics, have recently been considered as alternatives for methamphetamine abuse in Korea. In addition, the misuse and abuse of PT and PM obtained by illegal sources such as the internet become a serious social problem. In the present study, a simultaneous detection and quantification method for determining PT and PM in human urine was developed and validated according to the international guidelines. The urine samples were screened using a fluorescence polarization immunooassay and analyzed by gas chromatography mass spectrometry (GC-MS) after extraction using automatic solid phase extraction (SPE) with a mixed-mode cation exchange cartridge and derivatization with pentafluoropropionic anhydride (PFPA). The validation results for selectivity, linearity, limits of detection (LOD) and quantification (LOQ), intra- and inter-assay precision and accuracy and recovery were satisfactory. The validated method was successfully applied to authentic urine samples collected from 38 drug abuse suspects. PT and/or PM were identified with or without methamphetamine in urine samples. Abuse of PT and PM have increased continuously in Korea, therefore, closer supervision of the inappropriate use of anoretics is necessary.

• 약학회지
• /
• 제55권1호
• /
• pp.64-68
• /
• 2011

In this study, a high performance liquid chromatography-diode array detector method was established, for simultaneous determination of three compounds, berberine, palmatine and geniposide in Hwangryunhaedok-tang, To develop and validate method, $C_{18}$ column (5 ${\mu}M$, 4.6 mm${\times}$250 mm) was used with gradient mobile phase, water containing 0.1% trifluoroacetic acid (TFA) and MeOH at the column temperature of $30^{\circ}C$. UV wavelength was set at 230 and 280 nm. Validation of the chromatography method was evaluated by linearity, precision and accuracy test. Calibration curve of standard components showed good linearity ($R^2$ > 0.9999). The limits of detection (LOD) and limits of quantification (LOQ) varied from 0.05 to 0.17 ${\mu}g/ml$ and 0.15 to 0.53 ${\mu}g/ml$, respectively. The relative standard deviations (RSDs) data of intra-day and inter-day test were in less than 2.99% and 1.90%, respectively. The results of the accuracy test were in the range of 98.36 to 102.52% with RSDs values 0.32 to 1.98%. The results of validation indicated that this method was a very accurate and sensitive assay.

• Advances in Traditional Medicine
• /
• 제10권1호
• /
• pp.21-28
• /
• 2010

The objective of the present investigation is to evaluate the antimicrobial potency of the terpenoid fractions isolated from Luffa cylindrica seeds against various pathogenic microbes. The seeds were powdered and extracted with methanol in soxhlet appratus based on phytochemical screening. Three terpenoid components were isolated by column chromatography and identified by thin layer chromatography and chemical analysis which were designated as ${LCSF_4}^*$, ${LCSF_6}^*$ & ${LCSF_8}^*$ respectively. Disc diffusion method was employed to determine the antimicrobial effectiveness of test compounds I, II and III $({LCSF_4}^*,\;{LCSF_6}^*\;&\;{LCSF_8}^*)$ against 6 microbial species viz., Staphylococcus (S.) aureus, Bacillus (B.) subtilis, Escherichia (E.) coli, Pseudomonas (P.) aeruginosa, Candida (C.) albicans and Aspergillus niger. The disc was saturated with $100{\mu}l$ of each compound, allowed to dry and introduced on the upper layer of seeded agar plate. The plates were incubated overnight at $37^{\circ}C$. Microbial growth was determined by measuring the zonal inhibition diameters. Compound I showed maximum potency against gram positive S. aureus (21 mm) in comparison with standard ciprofloxacin (38 mm), whereas the same compound was completely devoid of activity against both the fungi tested. Compound II was found to be highly sensitive against both the gram negative E. coli (20 mm) and P. aeruginosa (22 mm). Compound II was found to exhibit maximum potency against the fungi C. albicans (15 mm) and A. niger (20 mm). Compound III was found to be very effective against both the gram positive S. aureus (20 mm) and B. subtilis (15 mm) respectively.

• Archives of Pharmacal Research
• /
• 제20권1호
• /
• pp.13-16
• /
• 1997

The cytotoxicity of animal venoms (snakes, insects and marine animals) was measured against SNU-1 (stomach cancer cells) by dye uptake assay (MTT method). And also L-amino acid oxidase (AAO) activity of the venoms was compared. Among them, the venom from Ophiophagus hannah (king cobra) showed a strong AAO activity as well as a high potent cytotoxicity. Cytotoxic protein having a AAO was then partially purified by HPLC-GPC and two fractions (Fr. I and Fr. II) were collected. The $IC_{50}$ values of Fr. I and Fr. II were 0.19 ${\mu}g/ml$ and 1.36 ${\mu}g/ml$, respectively. The results suggested that the cytotoxicity of king cobra venom may be due to its AAO activity.

• 약학회지
• /
• 제55권4호
• /
• pp.289-294
• /
• 2011

Illegal addition of steroids into cosmetics, ointments or drugs have been increased and their careless usage induced detrimental effect on health. We developed simultaneous analytical method using TLC, HPLC and LC/MS for the identification of 40 corticosteroids. 34 corticosteroids were well separated in HPLC with isocratic mode and remaining 6 drugs were also separated with gradient mode. All of the 40 corticosteroids were detected in negative mode in LC/MS. Halcinonide, prednisolone, triamcinolone acetonide and methylprednisolone hemisuccinate were detected in real samples.

• 한국산학기술학회논문지
• /
• 제12권3호
• /
• pp.1270-1277
• /
• 2011

제약산업은 연구개발 비중이 높은 지식기반산업으로 우수한 연구개발 인력에 기초한 연구 역량이 국가경쟁력의 핵심요소로 작용한다. 연구개발 역량이 우수한 국가가 세계시장을 선도하고, 높은 기술력만 있으면 세계 시장지배를 통해 고도성장이 가능한 분야다. 특히 제약분야의 연구개발 역량 강화를 위해서는 적정 규모의 연구인력 확보가 선결되어야할 과제다. 본 연구는 전문가 델파이 기법을 이용하여 제약분야의 연구개발 인력 수급 현황(2007년)과 함께 미래(2017년)의 인력수급 현황에 대해서 조사 분석하였다. 본 연구의 결과를 정리하면 다음과 같다. 첫째, 2007년 현재 제약 산업 분야의 연구개발 인력은 적정규모에 비해 약 5,600명 부족한 것으로 평가된다(부족률: 약 18.1%). 둘째, 향후 2017년에는 필요 연구개발 인력 대비 공급이 약 13,500명 부족할 것으로 전망되어 연구인력 수급 불균형은 향후 더 심화될 것으로 전망된다. 본 연구결과는 정부의 제약 분야에 대한 R&D 인력수급 정책의 변화가 없을 경우 향후 인력 수급에 심각한 차질(부족 현상)이 예상됨을 보여주는 만큼 합리적이고 체계적인 연구인력 수급 정책마련이 시급하게 필요함을 시사해 주고 있다.