• Journal of Pharmaceutical Investigation
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• v.37 no.3
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• pp.179-186
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• 2007

A simple, sensitive and selective liquid chromatographic/tandem mass spectrometric method (LC-MS/MS) was developed and validated for doxifluridine and 5-fluorouracil (5-FU) quantification in dog heparinized plasma. Sample preparation was based on liquid-liquid extraction using a mixture of isopropanol/ethyl acetate (1/9 v/v) to extract doxifluridine, 5-FU and 5-chlorouracil (5-CU, an internal standard) from plasma. Chromatography was performed on a C-18 analytical column and the retention times were 2.7, 1.5 and 1.7 min for doxifluridine, 5-FU and 5-CU, respectively with shorter analysis time within 5 min than previously reported methods. The ionization was optimized using ESI negative mode and selectivity was achieved by tandem mass spectrometric analysis by multiple reaction monitoring (MRM) using the transformations of m/z 244.8>107.6, 129.0>42.0 and 144.9>42.1 for doxifluridine, 5-FU and 5-CU, respectively. The achieved low limit of quantification was 20.0 ng/mL and the assay exhibited linear range of 20-2000 ng/mL ($R^2>0.99957$ for doxifluridine and $R^2>0.99857$ for 5-FU), using $100{\mu}L$ of plasma. Accuracy and precision of quality control samples for both doxifluridine and 5-FU met KFDA and FDA Guidance criteria of 15% for accuracy with coefficients of variation less than 15%. This method demonstrated adequate sensitivity, specificity, accuracy, precision and stability to support the simultaneous analysis of doxifluridine and 5-FU in dog plasma samples in pharmacokinetic and bioequivalence studies.

• Analytical Science and Technology
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• v.33 no.1
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• pp.23-32
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• 2020

Methamphetamine (MA) is the most common and available drug of abuse in Korea and its primary metabolite is amphetamine (AP). Detection of AP derivatives, such as MA, AP, phentermine (PT), MDA, MDMA, and MDEA by the use of immunoassay screening is not reliable and accurate due to cross-reactivity and insufficient specificity/sensitivity. Therefore, the analytical process accepted by most urine drug-testing programs employs the two-step method with an initial screening test followed by a more specific confirmatory test if the specimen screens positive. In this study, a gas chromatography-mass spectrometric (GC-MS) method was developed and validated for confirmation of MA and AP in human urine. Urine sample (500 µL) was added with N-isopropylbenzylamine as internal standard and ethyl chloroformate as a derivatization reagent, and then extracted with 200 µL of ethyl acetate. Extracted samples were analysed with GC-MS in the SIM/ Scan mode, which were screened by Cobas c311 analyzer (Roche/Hitachi) to evaluate the efficiency as well as the compatibility of the GC-MS method. Qualitative method validation requirements for selectivity, limit of detection (LOD), precision, accuracy, and specificity/sensitivity were examined. These parameters were estimated on the basis of the most intense and characteristic ions in mass spectra of target compounds. Precision and accuracy were less than 5.2 % (RSD) and ±14.0 % (bias), respectively. The LODs were 3 ng/mL for MA and 1.5 ng/mL for AP. At the screening immunoassay had a sensitivity of 100% and a specificity of 95.1 % versus GC-MS for confirmatory testing. The applicability of the method was tested by the analysis of spiked urine and abusers' urine samples.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.3
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• pp.161-165
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• 2004

Exogenous carbon monoxide (0.2%) increases L-type calcium $(Ca^{2+})$ current in human jejunal circular smooth muscle cells. The stimulatory effect of carbon monoxide (CO) on L-type $Ca^{2+}$ current is inhibited by pre-application of L-NNA, a classical competitive inhibitor of nitric oxide synthase (NOS) with no significant isoform selectivity (Lim, 2003). In the present study, we investigated which isoform of NOS affected CO induced stimulation of L-type $Ca^{2+}$ current in human jejunal circular smooth muscle cells. Cells were voltage clamped by whole-cell mode patch clamp technique, and membrane currents were recorded with 10 mM barium as the charge carrier. Before the addition of CO, cells were pretreated with each inhibitor of three NOS isoforms for 15 minutes. CO-stimulating effect on L-type $Ca^{2+}$ current was partially blocked by N-(3-(Amino-methyl) benzyl) acetamidine 2HCl (1400W, an iNOS inhibitor). On the other hand, 3-bromo-7-nitroindazole (BNI, a nNOS inhibitor) or $N^5-(1-Iminoethyl)-L-ornithine$ dihydrochloride (L-NIO, an eNOS inhibitor) completely blocked the CO effect. These data suggest that low dose of exogenous CO may stimulate all NOS isoforms to increase L-type $Ca^{2+}$ channel through nitric oxide (NO) pathway in human jejunal circular smooth muscle cells.

• Korean journal of applied entomology
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• v.31 no.3
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• pp.314-327
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• 1992

Insecticide resistance is a serious is a serious threat to IPM, resulting in various adverse effects not to mention the loss of yield in agriculture. One approach to counter the problem is the disruption of resistance mechanisms. This can be achieved by (1) compounds which show a negative correlation with resistance at the site of action, (2) specific metabolic inhibitors which serve as synergists, or (3) a certain combination of two insecticides producing a joint action. This approach, however, requires certain precautions for the side effects may cause an increase in toxicity to mammals. Owing to the recent advances in theoretical studies on resistance management employing computer simulation and mathematical models, a few principles to reduce the risk of development of resistance have been clarified. They are helpful in designing operational strategies with regard to, for instance, insecticide doses to be applied, mode of application, and choice and nature of the insecticide(s) to be used. For restoration of insecticide susceptibility of a resistant population, reintroduction of susceptible individuals to the resistant population is feasible when certain conditions are met. Natural enemies which developed resistance to insecticides can be an important component of IPM as has been shown in the pest management in apple orchards. After all, the implementation of a successful resistance management program depends upon cooperation between different sigments of the agricutural community. Although resistance is a preadaptive phenomenon, in some cases spontaneous loss of resistance does occur without contamination by susceptible individuals. The instability of resistance in these insects implies the possible existence of a switch machanism controlling the expression of resistance gene(s). Elucidation of such a mechanism may eventually provide us with a new technical approach with which we can combat the problem of insecticide resistance.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.5
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• pp.3191-3196
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• 2013

Leaf extracts of Cassia alata L (akapulko), traditionally used for treatment of a variety of diseases, were evaluated for their potential antitumor properties in vitro. MTT assays were used to examine the cytotoxic effects of crude extracts on five human cancer cell lines, namely MCF-7, derived from a breast carcinoma, SK-BR-3, another breast carcinoma, T24 a bladder carcinoma, Col 2, a colorectal carcinoma, and A549, a nonsmall cell lung adenocarcinoma. Hexane extracts showed remarkable cytotoxicity against MCF-7, T24, and Col 2 in a dose-dependent manner. This observation was confirmed by morphological investigation using light microscopy. Further bioassay-directed fractionation of the cytotoxic extract led to the isolation of a TLC-pure isolate labeled as f6l. Isolate f6l was further evaluated using MTT assay and morphological and biochemical investigations, which likewise showed selectivity to MCF-7, T24, and Col 2 cells with $IC_{50}$ values of 16, 17, and 17 ${\mu}g/ml$, respectively. Isolate f6l, however, showed no cytotoxicity towards the non-cancer Chinese hamster ovarian cell line (CHO-AA8). Cytochemical investigation using DAPI staining and biochemical investigation using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-a method used to detect DNA fragmentation-together with caspase assay, demonstrated apoptotic cell death. Spectral characterization of isolate f6l revealed that it contained polyunsaturated fatty acid esters. Considering the cytotoxicity profile and its mode of action, f6l might represent a new promising compound with potential for development as an anticancer drug with low or no toxicity to non-cancer cells used in this study.

• Analytical Science and Technology
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• v.27 no.6
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• pp.339-346
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• 2014

The author assessed the availability of urine reference material for proficiency test provided for laboratories in occupational health. N-methylacetamide is the biomarker of exposure to N,N-dimethyl acetamide, which was used as the substitute for hepatotoxic N,N-dimethylformamide (DMF). N-methylacetamide (NMAC) urine samples of 3 different levels covering the 0.2~2 times of the exposure limit were tested. Stability test up to 180 days (0, 7, 30, 60, 180 days) at 4 different temperatures (-60, -20, 5, $25^{\circ}C$) and homogeneity test were performed for these samples. New analytical condition by GC/MSD using SIM mode (m/z 58, 59) and DB-624 column was investigated for better selectivity, sensitivity and resolution. Urinary NMAC samples showed good homogeneity for 3 levels. These samples also showed good stability up to 180 days. The data of stability and homogeneity of urinary DMAC confirmed the basis of including this item into Korean proficiency test for occupational health laboratories since 2008.

• Biomolecules & Therapeutics
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• v.16 no.4
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• pp.336-342
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• 2008

The resveratrol analogue piceatannol (3,5,3',4'-tetrahydroxy-trans-stilbene) is a polyphenol present in grapes and wine and reported to have anti-carcinogenic activities. To investigate the mechanism of anticarcinogenic activities of piceatannol, the effects on CYP 1 enzymes were determined in Escherichia coli membranes coexpressing recombinant human CYP1A1, CYP1A2 or CYP1B1 with human NADPH-P450 reductase. Piceatannol showed a strong inhibition of CYP1A1 and CYP1B1 in a concentration-dependent manner, and $IC_{50}$ of human CYP1A1 and CYP1B1 was 5.8 ${\mu}M$ and 16.6 ${\mu}M$, respectively. However, piceatannol did not inhibit CYP1A2 activity in the concentration of up to 100 ${\mu}M$. Piceatannol exhibited 3-fold selectivity for CYP1B1 over CYP1A1. The mode of inhibition of piceatannol was non-competitive for CYP1A1 and CYP1B1. The result that piceatannol did not inhibit CYP1B1-mediated $\alpha$-naphthoflavone ($\alpha$-NF) metabolism suggests piceatannol may act as a non-competitive inhibitor as well. In human prostate carcinoma PC-3 cells, piceatannol induces apoptosis and prevents Aktmediated signal pathway. Taken together, abilities of piceatannol to induce apoptotic cell death as well as CYP1 enzyme inhibition make this compound a useful tool for cancer chemoprevention.

• Journal of the Korean Society for Nondestructive Testing
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• v.30 no.4
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• pp.373-379
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• 2010

EMAT has been applied in various fields for flaw detection and material characterization because it has noncontact property in wave generation and a good mode selectivity. Unfortunately, however, EMAT shows low signal to noise ratio relative to commercial contact transducer because of low energy conversion efficiency. If the phase matching through the control of time delay between each coil consisting of the array EMAT is accomplished, it is expected that it will be a solution for the improvement of low signal to noise ratio. In this experiment, the phased array EMATs which consists of 3 or 4 meander coils and one big magnet were fabricated for surface and vertical shear wave generation. Effect of phased delay control on signal directivity and amplitude enhancement was verified. A slit with the depth of 0.5 mm and a side-drill hole of 0.5 mm diameter were clearly detected by fabricated phased array EMATs, respectively.

• 한국해양학회지
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• v.21 no.4
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• pp.236-244
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• 1986

This study was carried out for appreciable information on the proper habitats and feeding modes of some dominant polychaetes in the continental shelf of the East Sea during April, 1985. Among 95 polychaete species, we select ed 9 dominant species in terms of their occurring frequency, abundance and population density. These are Terebellides stroemi, Chaetozone setosa, Magelona japonica, Ampharete arctica, Aglaophamus sinensis, Nothria holobranchiata, Lumbrineris japonica, Myriochele gracilis, Notoproctus pacificus. Major food item s of these species are centric diatoms and detritus. The feeding modes of the the m are mainly surtace or subsurface deposit feeding, though two specters, M.holobranchiata and L. japonica, could be assumed to be potential carnivores. To a certain degree, most deposit feeders in the coarse sediments showed selectivity in feeding and tube building. The feeding mode, gut content and the tube structure, and the distribution pattern of dominant polychaetes correspond well with the habitat tharacters, e.g.the bottom topography, the source of food and the sediment composition.

• Bulletin of the Korean Chemical Society
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• v.34 no.11
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• pp.3284-3288
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• 2013

This paper describes first development and validation of pentafluorophenylprophyl ligand-based liquid chromatography coupled to tandem mass spectrometry (PFPLC-MS/MS) method to determine metformin, a highly polar compound, in human plasma. Metformin and Phenformin (internal standard) were extracted from human plasma 50 ${\mu}L$ with a single-step protein precipitation. The chromatographic separation was performed using a linear gradient elution of mobile phase involving 5.0 mM ammonium formate solution with 0.1% formic acid (A) and acetonitrile (B) over 3.0 min of run time on a Phenomenex Luna PFP column. The detection was performed using a triple-quadrupole tandem mass spectrometer (Waters Quattro micro) with electrospray ionization in the mode of positive ionization and multiple-reaction monitoring (MRM). The developed method was validated with 5.0 ng/mL of lower limit of quantification (LLOQ). The calibration curve was linear over 5-3000 ng/mL of the concentration range ($R^2$ > 0.99). The specificity, selectivity, carry-over effect, precision, accuracy and stability of the method met the acceptance criteria. The method developed in this study had had rapidness, simplicity and ruggedness. The reliable method was successfully applied to high throughput analysis of real samples for a practical purpose of a pharmacokinetic study.