• Archives of Pharmacal Research
• /
• v.27 no.9
• /
• pp.901-905
• /
• 2004

A rapid, sensitive and selective hydrophilic interaction liquid chromatography-tandem mass spectrometric(HILIC-MS/MS) method for the determination of tiapride in human plasma was developed. Tiapride and internal standard, metoclopramide were extracted from human plasma with dichloromethane at basic pH and analyzed on an Atlantis HILIC silica column with the mobile phase of acetonitrile-ammonium formate (190 mM, pH 3.0) (94：6, v/v). The ana-Iytes were detected using an electrospray ionization tandem mass spectrometry in the multi-ple-reaction-monitoring mode. The standard curve was linear (r＝0.999) over the concentration range of 1.00-200 ng/mL. The coefficient of variation and relative error for intra- and inter-assay at three QC levels were 6.4∼8.8％ and -2.0∼3.6％, respectively. The recoveries of tiapride ranged from 96.3 to 97.4％, with that of metoclopramide (internal standard) being 94.2％. The lower limit of quantification for tiapride was 1.00 ng/mL using 1 00 $\mu$L of plasma sample.

• YAKHAK HOEJI
• /
• v.54 no.4
• /
• 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.

• Proceedings of the KWS Conference
• /
• 2002.10a
• /
• pp.53-58
• /
• 2002

Electrical insulation and mechanical properties of the plasma sprayed oxide ceramic coatings were studied before and after the sealing treatment of the ceramic coatings. Plasma sprayed A1$_2$O$_3$-TiO$_2$ coating as the reference coating was sealed using three commercial sealants based on polymer. Penetration depth of the sealants to the ceramic coating was evaluated directly from the optical microscope using a fluorescent dye. It is estimated that the penetration depth of the sealants to the ceramic coating is from 0.2 to 0.5 mm depending on the sealants used. The preliminary test results with a DC puncture tester imply that the dielectric breakdown voltage mechanism of plasma sprayed ceramic coatings has been determined to be a corona mechanism. Dielectric breakdown voltage of the as-sprayed and as-ground samples have shown a linear trend with regard to the thickness showing an average dielectric strength of 20 kV/mm for the thickness scale studied. It is also shown that grinding the coating before sealing and adding fluorescent dye do not agent the penetration depth of sealants. All of the microhardness, two-body abrasive wear resistance, bond strength, and surface roughness of the ceramic coating after the sealing treatment are improved. The extent of improvement is different from the sealants used. However, three-point bending stress of the ceramic coating after the sealing treatment is decreased. This is attributed to the reduced micro-crack toughening effect since the cracks propagate easily through the lamellar of the coating without crack deflection and/or branching after the sealing treatment.

• Biomolecules & Therapeutics
• /
• v.15 no.4
• /
• pp.266-272
• /
• 2007

A rapid analytical method was developed to quantify very long-chain fatty acids (VLCFAs, C22:0, C24:0, C26:0) in human plasma with good sensitivity and specificity using tert-butyldimethylsilyl (TBDMS) derivatization and stable isotope GC-MS selective ion monitoring (GC-MS/SIM). Two-hundred and fifty ${\mu}L$ of plasma was fortified with deuterated stable isotope internal standards (d3-C22:0, d3-C24:0, d3-C26:0) and standard mixtures of chloroform and methanol, and then extracted with hexane and acetonitrile. To upper layer of liquid-liquid-extraction, N-(t-Butyldimethylsilyl)-N-methyltrifluoroacetamide was added and then heated to $60^{\circ}C$ for 30 min to produce the TBDMS derivatives. Derivatives of VLCFAs were analyzed by GC-MS/SIM. Calibration curves showed a linear relationship for the target compounds in the concentration range of $10^{-4}{\sim}2{\times}10^3\;{\mu}g/mL$ with the correlation coefficient ranging from 0.996 to 0.999. The limit of quantification for the plasma was $10^{-4}{\sim}2{\times}10^{-4}\;{\mu}g/mL$ (S/N=3). When applied to the plasma specimens of patients with peroxisomal disorder, X-linked adrenoleucodystropy (ALD, Mckusick 202370), the method clearly differentiated normal subjects from ALD patients. The C24:0/C22:0 and C26:0/C22:0 ratios were significantly elevated in the plasma of patients with X-linked ALD compared to normal subjects. The new developed method might be useful for a rapid and sensitive diagnosis of X-linked ALD and other peroxisomal disorders.

• Bulletin of the Korean Chemical Society
• /
• v.23 no.8
• /
• pp.1139-1143
• /
• 2002

Analysis of lysophosphatidic acids (LPAs) is of clinical importance as they can serve a potential marker for ovarian and other gynecological cancers and obesity. It is critically important to develop a highly sensitive and specific method for the early detection of gynecological cancers to improve the overall outcome of this disease. We have established a novel quantification method of LPAs in human plasma by negative ionization tandem mass spectrometry (MS-MS) using multiple reaction monitoring (MRM) mode without the conventional TLC step. Protein-bound lipids, LPAs in plasma were extracted with methanol : chloroform (2:1) containing LPA C14:0 as an internal standard under acidic condition. Following back extraction with chloroform and water, the centrifuged lower phase was evaporated and reconstituted in methanol. The reconstituted solution was directly injected into electrospray source of MS/MS. For MRM mode, Q1 ions selected were m/z 409, 433, 435, 437 and 457 which corresponds to molecular mass [M-H]- of C16:0, C18:2, C18:1, C18:0 and C20:4 LPA, respectively. Q2 ions selected for MRM were m/z 79, phosphoryl product. Using MS/MS with MRM mode, all the species of LPAs were completely separated from plasma matrix without severe interferences. This method allowed simultaneous detection and quantification of different species of LPAs in a plasma over a linear dynamic range of 0.01-25 ㎛olL-1 . The detection limit of the method was 0.3 pmol/mL, with a correlation coefficient of 0.9983 in most LPAs analyzed. When applied to the plasmas of normal and gynecological cancer patients, this new method differentiated two different groups by way of total LPA level.

• Preventive Nutrition and Food Science
• /
• v.19 no.4
• /
• pp.321-326
• /
• 2014

In this study, we developed a method to quantify esculetin (6,7-dihydroxycoumarin) in plasma and tissues using HPLC coupled with ultraviolet detection and measured the level of esculetin in rat plasma after oral administration. The calibration curve for esculetin was linear in the range of 4.8 ng/mL to 476.2 ng/mL, with a correlation coefficient ($r^2$) of 0.996, a limit of detection value of 33.2 ng/mL, and a limit of quantification value of 100.6 ng/mL. Recovery rates for the 95.2 ng/mL and 190.5 ng/mL samples were 95.2% and 100.3%, within-runs and 104.8% and 101.0% between-runs, respectively. The relative standard deviation was less than 7% for both runs. In the pharmacokinetic analysis, the peak plasma esculetin level was reached 5 min after administration ($C_{max}=173.3ng/mL$; $T_{1/2}=45min$; $AUC_{0{\sim}180min}=5,167.5ng{\cdot}min/mL$). At 180 min post-administration (i.e., after euthanasia), esculetin was only detectable in the liver ($30.87{\pm}11.33ng/g$) and the kidney ($20.29{\pm}7.02ng/g$).

• Journal of Animal Science and Technology
• /
• v.47 no.1
• /
• pp.29-38
• /
• 2005

Effect of dietary brown seaweed(Undaria pinnatifida) levels on the anti-oxidant enzyme system was evaluated in blood of broiler chicks activated innate immune response. Day-old broiler chicks were fed diets containing 0.0(basal), 1.0, 2.0 and 4.0 % of brown seaweed for 4 weeks. The innate immune response was activated by injecting Salmonella typhymurium lipopolysaccharide(LPS) i.p. at 8, 10 and 12 day of age. The activation of innate immune response enhanced(p< 0.01) and the brown seaweed 1.0 and 2.0 % diets reduced(P< 0.05) the superoxide dismutase(SOD) activity in erythrocyte cytosol significantly. The activation of innate immune response elevated significantly the levels of peroxide and the activity of peroxidase in plasma, while the levels of dietary brown seaweed resulted in a significant linear increase in peroxidase activity in plasma of normal bird. Experience of the innate immune response in 4 week-old chicks reduced linearly the plasma level of peroxide with the level of brown seaweed and enhanced the plasma peroxidase activity. Also, the plasma of normal birds fed the brown seaweed showed higher level of peroxide and lower activity of peroxidase. The results indicated that dietary brown seaweed affected SOD and peroxidase activities in blood of broiler chicks during activation of innate immune response.

• Archives of Pharmacal Research
• /
• v.29 no.4
• /
• pp.339-342
• /
• 2006

A simple high performance liquid chromatographic (HPLC) method was developed for the determination of tolperisone in human plasma. Tolperisone and internal standard (chlorphenesin) were isolated from 1 mL of plasma using 8 mL of dichlormethane. The organic phase was collected and evaporated under nitrogen gas. The residue was then reconstituted with 300 mL aliquot of mobile phase and a 100 mL aliquot was injected onto the $C_{18}$ reverse-phased column. The mobile phase, $45\%$ methanol containing $1\%$ glacial acetic acid and $0.05\%$ 1-hexanesulfonic acid was run at a flow rate of 1 mL/min. The column effluent was monitored using UV detector at 260 nm. The retention times for tolperisone and the internal standard were approximately 7.1 and 8.4 min, respectively. The standard curve was linear with minimal intra-day and inter-day variability. The quantification limit of tolperisone in human plasma was 10 ng/ mL. The proposed method has been applied to the determination of pharmacokinetic profile of tolperisone in Koreans. The T max of tolperisone in Koreans $(0.94{\pm}0.42\;h)$ was not significantly differ from that reported in Europeans (0.5-1 h), but the mean half-life in Koreans $(1.14{\pm}0.27\;h)$ was shorter than that in Europeans $(2.56{\pm}0.2\;h)$. The proposed HPLC method is simple, accurate, reproducible and suitable for pharmacokinetic study of tolperisone.

• Toxicological Research
• /
• v.29 no.3
• /
• pp.203-209
• /
• 2013

A simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of ${\varepsilon}$-acetamidocaproic acid (AACA), the primary metabolite of zinc acexamate (ZAC), in rat plasma by using normetanephrine as an internal standard. Sample preparation involved protein precipitation using methanol. Separation was achieved on a Gemini-NX $C_{18}$ column ($150mm{\times}2.0mm$, i.d., 3 ${\mu}m$ particle size) using a mixture of 0.1% formic acid-water : acetonitrile (80 : 20, v/v) as the mobile phase at a flow rate of 200 ${\mu}l/min$. Quantification was performed on a triple quadrupole mass spectrometer employing electrospray ionization and operating in multiple reaction monitoring (MRM) and positive ion mode. The total chromatographic run time was 4.0 min, and the calibration curves of AACA were linear over the concentration range of 20~5000 ng/ml in rat plasma. The coefficient of variation and relative error at four QC levels were ranged from 1.0% to 5.8% and from -8.4% to 6.6%, respectively. The present method was successfully applied for estimating the pharmacokinetic parameters of AACA following intravenous or oral administration of ZAC to rats.

• The Transactions of the Korean Institute of Power Electronics
• /
• v.18 no.6
• /
• pp.593-600
• /
• 2013

The asymmetric pulsed DC reactive magnetron sputtering system is widely used for the high quality plasma sputtering process such as a thin film deposition. In asymmetric pulsed DC power supply a reverse voltage is applied to the target periodically to minimize arc discharging effect. When sputtering in the mid-frequency range (20-350 kHz), the periodic target voltage reversals suppress arc formation at the target and provide long-term process stability. Thus, high quality, defect-free coatings of these materials can now be deposited at competitive rates. In this paper, a new style asymmetric pulsed DC power supply including mid-transformer is presented. In the proposed, an energy recovery circuit is adopted to reduce the mutual inductance of the transformer. As a result, the system dynamics of the voltage control loop is increased highly and the non-linear voltage boosting effect of the conventional system is removed. This work was proved through simulation and laboratory based experimental study.