• Mass Spectrometry Letters
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• 제9권4호
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• pp.95-99
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• 2018

An innovative, simple, and rapid assay method based on liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was developed and validated for the simultaneous determination of eight statin drugs in human urine. A simple sample clean-up procedure using the "dilute and shoot" (DAS) approach enabled a fast and reliable analysis. The influence of the dilution factor was investigated to ensure detectability and reduce the matrix effect. Chromatographic separation was performed on a Phenomenex Kinetex C18 column ($50{\times}3.0mm$ i.d., $2.6{\mu}m$) using an elution gradient of mobile phase A composed of 0.1% acetic acid, and mobile phase B composed of acetonitrile, at a flow rate of 0.35 mL/min. Quantitation was performed on a triple quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) mode using electrospray ionization in positive ion mode. The total chromatographic run time was 15 min. The method was validated for selectivity, sensitivity, recovery, linearity, accuracy, precision, and stability. The present method was successfully applied to the analysis of Rosuvastatin in urine samples after oral administration to healthy human subjects.

• 생약학회지
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• 제50권3호
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• pp.198-204
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• 2019

A facile and convenient method was developed for the mass production of epigallocatechin gallate (EGCG) rich green tea extract (Er-GTE). The Er-GTE was successfully obtained from the crude water extract of green tea by the combination of two step purification, i.e., a simple adsorption process on the cation exchange resins (Trilite SCR-B) followed by the chromatography with Diaion HP-20 resins. The green tea extract produced by water extraction under $45^{\circ}C$ was subjected to adsorb on the strongly acidic cation exchange resin, Trilite SCR-B. The eluate passed through the resin was reabsorbed on Diaion HP-20 resin, which was subjected to elute with a mixture of water and alcohol by conventional chromatographical manner. The EGCG content in Er-GTE was estimated above 97% by HP-LC analysis and the newly developed method was regarded as the most suitable and appropriate process for the mass production of epigallocatechin gallate rich green tea extract (Er-GTE).

• 한국해양바이오학회지
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• 제14권2호
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• pp.143-154
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• 2022

We isolated marine actinomycetes, strain D-5 which produces anti-methicillin resistant Staphylococcus aureus (anti-MRSA) compound. Streptomyces sp. D-5 relatively grew well in the 20~25℃, pH 8.0, and NaCl 3.0%. The ethyl acetate extract of D-5 culture was separated by C₁₈ ODS open column and reverse phase HPLC to yield anti-MRSA compound. The molecular weight of this compound was determined to be 898 by a Liquid chromatograph-mass spectrometer (LC-MS). Compared with penicillin G, this compound showed significant anti-MRSA activity. It also exhibited an inhibition zone of 26 mm at a concentration of 64 ㎍/disk and an inhibition zone of 16 mm at a concentration of 16 ㎍/disk against the MRSA KCCM 40511. Furthermore, the co-treatment of HPLC peak 5 compound and vancomycin caused a more rapid decrease in MRSA cells than each compound alone. It showed 86.8% growth inhibition activity within 12 hours at a low concentration of 50 ㎍/mL during co-treatment, and 97.1% growth in-hibition activity within 48 hours against MRSA KCCM 40511. Taken together, our results suggest that Streptomyces sp. D-5 and its anti-MRSA compound could be employed as a potent agent in MRSA infection.

• 분석과학
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• 제36권6호
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• pp.281-290
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• 2023

Zidovudine is an antiretroviral agent prescribed for the prevention and treatment of human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS). It is typically recommended to be used in combination with other antiretroviral drugs. Zidovudine has the potential to generate N-nitrosodimethylamine (NDMA) in the presence of dimethylamine and nitrite salt under acidic reaction conditions during the drug manufacturing process. NDMA is a potent human carcinogen that may be detected in drug substances or drug products. An analytical method was developed to determine NDMA in pharmaceuticals including zidovudine using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The analysis involved reversed-phase chromatography on a Kinetex F5 column with a mobile phase comprising water-acetonitrile mixtures. The detection of positively charged ions was conducted using atmospheric pressure chemical ionization (APCI). The calibration curve demonstrated excellent linearity (r = 0.9997) across the range of 1-50 ng/mL with a highly sensitive limit of detection (LOD) at 0.3 ng/mL. The developed method underwent thorough validation for specificity, linearity, accuracy, precision, robustness, and system suitability. This sensitive and specific analytical method was applied for detecting NDMA in zidovudine drug substance and its formulation currently available in the market, indicating its suitability for drug quality management purposes.

• 농약과학회지
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• 제6권1호
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• pp.31-35
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• 2002

본 논문의 목적은 benfuracarb 원제(90.2%)에 함유된 불순물의 glutathione-S-transferase와 amidase에 대한 저해 특성과 해당 불순물의 구조를 밝히는데 있다. Benfuracarb 원제, 유효성분 및 불순물은 glutathione-S-transferase(GST)를 효과적으로 저해하였으나, 그 저해력은 GST 효소 저해제인 ethacrynic acid의 저해력보다는 낮았다. 즉, GST에 대한 benfuracarb 원제, 유효성분 및 불순물의 $I_{50}$은 각각 $9.7{\times}10^{-4}M,\;>1.0{\times}10^{-3}M,\;1.8{\times}10^{-4}M$이었으나, ethacrynic acid의 $I_{50}$은 $1.7{\times}10^{-5}M$이었다. Benfuracarb 원제, 유효성분 및 불순물은 amidase를 저해하였는데, 이들의 효소저해력은 iprobenfos의 저해력($I_{50},\;8.2{\times}10^{-7}M$)보다는 낮은 $6.0{\times}10^{-5}M,\;4.3{\times}10^{-4}M,\;7.6{\times}10^{-5}M$이었다. Benfuracarb 원제에는 4종의 불순물(IM $1{\sim}4$)이 검출되었는데, 이들 중 IM 2와 3은 GST와 amidase의 활성을 저해하였던 반면, IM 4는 효소활성을 저해하지 않았다. 이들 불순물 중 효소활성 저해특성을 갖는 IM 2와 3을 IR, $^1H$-NMR, $^{13}C$-NMR, LC-MS를 이용하여 구조를 분석한 결과, IM 2는 ethyl-N-isopropylamino propionate로, 그리고 IM 3은 ethyl-N-isopropyl-N-(chlorosulfenyl) aminopropionate로 확인되었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.39-39
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• 2019

섬초롱꽃은 울릉군에 한정적으로 분포하는 여러해살이풀이다. 산림청에서는 약관심종(LC) 등급의 희귀식물이자 특산식물로 분류하고 있으며, 꽃의 관상가치가 특히 높아 화단용 소재, 절화용, 화분재배 등으로 흔히 이용되고 있다. 따라서 본 연구는 섬초롱꽃의 대량증식 시, 중금속이 미칠 수 있는 영향에 대해 파악하고자 수행하였다. 실험에 사용한 중금속은 카드뮴(Cd), 철(Fe) 두 종류이며, 두 종류 모두 1% 한천(agar)배지에 10, 25, 50, 100, 200 mg/L 농도로 녹여 사용하였다. 발아율 및 유근 생장을 위해 종자 및 유묘를 치상 후, $25^{\circ}C$ 항온 생장상에서 발아율 및 유근 발달을 관찰하였다. 발아율 측정결과, 아무처리도하지 않은 1% 한천(agar) 배지에서 $62.7{\pm}7.4%$로 가장 높았고 중금속 농도가 높아질수록 감소하였다. 카드뮴(Cd) 처리구에서는 10, 25, 50, 100, 200 mg/L 농도에서 각각 $17.3{\pm}1.3$, $2.7{\pm}1.3$, $1.3{\pm}1.3$, 0, 0%로 나타났다. 한편, 철(Fe) 처리구에서는 10, 25, 50, 100, 200 mg/L 농도에서 각각 $46.7{\pm}4.8$, $24.0{\pm}2.3$, $36.0{\pm}6.9$, $20.0{\pm}2.3$, 0%로 나타났다. 유근의 평균 생장 길이(mm)는 아무처리도 하지 않은 1% 한천(agar) 배지에서 19.6 mm로 가장 높았고, 중금속 농도가 높아질수록 낮게 나타났다. 카드뮴(Cd) 처리구에서는 10, 25, 50, 100, 200 mg/L 농도에서 각각 4.6 mm, 1.9 mm, 0.7 mm, 0.9 mm, 0 mm 생장한 것으로 나타났다. 한편, 철(Fe) 처리구에서는 10, 25, 50, 100, 200 mg/L 농도에서 각각 15.9 mm, 8.6 mm, 8.1 mm, 0.9 mm, 0 mm 생장한 것으로 나타났다. 실험결과를 종합해 볼 때, 섬초롱꽃 종자 발아 및 유근 생장의 억제는 카드뮴(Cd) 처리구에서 높게 나타났고, 철(Fe) 처리구에서는 비교적 영향을 덜 미치는 것으로 나타났다. 특히 카드뮴은 10 mg/L 농도만 처리하더라도 약 3.6~4.2배의 발아율 생장율 감소가 나타나, 섬초롱꽃 대량증식 시, 치명적일 것으로 판단된다. 본 실험의 결과는 섬초롱꽃 종자를 이용한 대량증식 및 유묘재배에 있어, 토양을 선택하는데 중요한 기초자료로서 활용될 것으로 기대된다.

• International Journal of Industrial Entomology and Biomaterials
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• 제3권1호
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• pp.19-23
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• 2001

Two B. thuringiensis strains, which possess mosquitocidal activities, were isolated from Korean soil samples and named K-1205-1 and K-1381-1. Serological studies indicated that K-1205-1 and K-1381-1 belonged to B. thuringiensis subsp. kurstaki (H3a3b3c) and subsp. aizawai (H7), respectively. K-1205-1 produced typical bipyramidal parasporal inclusions, but K-1381-1 produced irregular bipyramidal shape. Total plasmid DNA patterns analysis shewed that K-1205-1 and K- 1381-1 were different from their reference strains, subsp. kurstaki and subsp. aizawai, respectively, in high molecules, whereas their crystal protein patterns showed no difference. The cry gene contents of K-1205-1 and K-1381-1 were identical with those of the reference strains. Mosquitocidal activities of crystal proteins produced by K-1205-1 and K-1381-1 were significantly high by about 40-50 folds at $LC_50$ when compared to those of subsp. kurstaki and subsp. aizawai. Finally, in southern blot analysis using cry1A-type specific probe, K-1205-1 and K-1381-1 had different bands from subsp. kurstaki and subsp. aizawai, respectively. In conclusion, our results suggest that K-1205-1 and K-1381-1 appear to be new moquitocidal B. thuringiensis strains isolated from Korean soil.

• Biomolecules & Therapeutics
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• 제21권2호
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• pp.114-120
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• 2013

Most patients with mutant B-Raf melanomas respond to inhibitors of oncogenic B-Raf but resistance eventually emerges. To better understand the mechanisms that determine the long-term responses of mutant B-Raf melanoma cells to B-Raf inhibitor, we used chronic selection to establish B-Raf (V600E) melanoma clones with acquired resistance to the new oncogenic B-Raf inhibitor UI-152. Whereas the parental A375P cells were highly sensitive to UI-152 ($IC_{50}$ < $0.5{\mu}M$), the resistant sub-line (A375P/Mdr) displayed strong resistance to UI-152 ($IC_{50}$ < $20{\mu}M$). Immunofluorescence analysis indicated the absence of an increase in the levels of P-glycoprotein multidrug resistance (MDR) transporter in A375P/Mdr cells, suggesting that resistance was not attributable to P-glycoprotein overexpression. In UI-152-sensitive A375P cells, the anti-proliferative activity of UI-152 appeared to be due to cell-cycle arrest at $G_0/G_1$ with the induction of apoptosis. However, we found that A375P/Mdr cells were resistant to the apoptosis induced by UI-152. Interestingly, UI-152 preferentially induced autophagy in A375P/Mdr cells but not in A375P cells, as determined by GFP-LC3 puncta/cell counts. Further, autophagy inhibition with 3-methyladenine (3-MA) partially augmented growth inhibition of A375P/Mdr cells by UI-152, which implies that a high level of autophagy may protect UI-152-treated cells from undergoing growth inhibition. Together, our data implicate high rates of autophagy as a key mechanism of acquired resistance to the oncogenic B-Raf inhibitor, in support of clinical studies in which combination therapy with autophagy targeted drugs is being designed to overcome resistance.

• Bulletin of the Korean Chemical Society
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• 제34권9호
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• pp.2760-2764
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• 2013

Geniposide is a biologically active ingredient of gardenia fruit. A liquid chromatography-tandem mass spectrometric method was developed and validated for the determination of geniposide in rat plasma. The plasma samples were pretreated by solid-phase extraction and introduced into a BDS Hypersil $C_{18}$ column ($50{\times}2.1mm$, $5{\mu}m$) for chromatographic separation. The mobile phase consisted of 0.1% formic acid and 0.1% formic acid in acetonitrile, and gradient elution was performed at a flow rate of 0.25 mL/min. For mass spectrometric detection, multiple reaction monitoring was performed via an electrospray ionization source in positive mode. The calibration curve for geniposide was linear ($r^2=0.997$) in the concentration range of $0.005-1{\mu}g/mL$. The intra- and inter-day accuracies and precisions fulfilled the required criteria (${\pm}15%$). The developed method was subsequently used for pharmacokinetic analysis of geniposide after oral administration to rats at a dose of 50 mg/kg. The mean maximum plasma concentration of geniposide was $0.68{\pm}0.29{\mu}g/mL$ at $0.44{\pm}0.13h$, and the mean area under the plasma concentration versus time curve was $1.46{\mu}g{\cdot}h/mL$.

• 동의생리병리학회지
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• 제16권2호
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• pp.262-266
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• 2002

In order to elucidate the mechanism of cytotoxic effect of oxygen radicals on cultured mouse spinal motor neurons, the neurotoxicity induced by hydrogen peroxide(H₂O₂) was evaluated by MTT assay. The neuroprotective effect of Rhizoma Gastrodiae(RG) against H₂O₂-mediated neurotoxicity was also examined in these cultures by SRB assay. The results were as follows : The value of lethal concentration 50(LC50) of H₂O₂ was estimated at a concentration of 30 uM in these cultures. Cell viability of cultured mouse spinal motor neurons was remarkably decreased by H₂O₂-induced neurotoxicity in a dose- and time-dependent manner. RG was remarkably effective in blocking the neurotoxicity induced by H₂O₂ at a concentration of 120 μM as determined by SRB assay. From above the results, it is suggested that H₂O₂ induce neurotoxicity, and the selective herbal extracted RG was very effective in blocking H₂O₂-mediated neurotoxicity on cultured mouse spinal motor neurons.