• BMB Reports
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• 제30권6호
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• pp.390-396
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• 1997

A sandwich-type enzyme-linked immunosorbent assay (ELISA) for the quantification of human apolipoprotein A-I (apoA-I) was developed using monoclonal antibodies. For this assay, we used three monoclonal antibodies to trap and detect apo A-I. HDAI16 and HDA15 monoclonal antibodies were used for trapping apoA-I and HDAI8 monoclonal antibody was for detecting apoA-I. These three monoclonal antibodies were produced by immunizing mice with high density lipoprotein (HDL) isolated from human plasma. By immunoblot analysis, these three monoclonal antibodies were specific to apoA-I and showed no cross-reactivities with other plasma proteins. The results of competition assays for epitope cross-reactivity test also verified that these monoclonal antibodies identified separate and distinct epitopes on HDL and apoA-I. Affinity constants of monoclonal antibodies were measured by ELISA. Their association constants ranged from $10^7$ to $10^8$ $M^{-1}$. For this assay, pure apoA-I was isolated by affinity chromatography using monoclonal antibodies. In this sandwich assay, the amount of HRP-labeled HDAI8 bound to apoA-I trapped by HDAI16 and HDAI5 was proportional to apoA-I concentration in the range of 0 to 500ng/ml. ApoA-I concentration in plasma was calculated from the linear regression equation of standard curve. The precision and reliability of the assays are reflected in the low intra-and interassay coefficients of variation that averaged 3.25% and 4.30%, respectively. This assay is sensitive, simple, reproducible, convenient in incubation interval, and does not use radioisotope: thus it can be widely applied in clinical laboratories.

• Natural Product Sciences
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• 제17권4호
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• pp.328-336
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• 2011

A high-performance liquid chromatography (HPLC) with diode array detector (DAD) method was established for the discrimination of a folk medicine Forsythia suspensa and Forsythia viridissima. Five and three representative metabolites of the lignan and phenolic glycoside classes were selected for the analysis from F. suspensa and F. viridissima, respectively. The optimal chromatographic conditions were obtained on an ODS column (5 ${\mu}m$, $4.6{\times}250$ mm) with the column temperature at $40^{\circ}C$. The mobile phase was composed of methanol and 0.3% acetic acid using an isocratic elution with the flow rate 1 mL/min. Detection wavelength was set at 280 nm. All calibration curves showed good linear regression ($r^2$ > 0.996) within test ranges. Limits of detection (LOD) and limits of quantitation (LOQ) values were lower than 0.096 and 0.291 ${\mu}g/mL$, respectively. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.07-0.63% and 0.14-0.62%, respectively, and the overall recoveries of 97.79-102.46% for all of the compounds analyzed. In addition, effectiveness of diverse extraction methods was compared to each other for the development of standard analytical method. The verified method was successfully applied to the quantitative determination of representative metabolites in fifty-three commercial F. suspensa samples and fifteen commercial F. viridissima samples from diverse sources. The overall analytical results showed the unequivocal differences in bioactive constituents between F. suspensa and F. viridissima.

• Toxicological Research
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• 제31권3호
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• pp.313-319
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• 2015

3-Monochloro-1,2-propanediol (3-MCPD) and 1,3-dichloro-2-propanol (1,3-DCP) are not only produced in the manufacturing process of foodstuffs such as hydrolyzed vegetable proteins and soy sauce but are also formed by heat processing in the presence of fat and low water activity. 3-MCPD exists both in free and ester forms, and the ester form has been also detected in various foods. Free 3-MCPD and 1,3-DCP are classified as Group 2B by the International Agency for Research on Cancer. Although there is no data confirming the toxicity of either compound in humans, their toxicity was evidenced in animal experimentation or in vitro. Although few studies have been conducted, free 3-MCPD has been shown to have neurotoxicity, reproductive toxicity, and carcinogenicity. In contrast, 1,3-DCP only has mutagenic activity. The purpose of this study was to analyze 3-MCPD and 1,3-DCP in various foods using gas chromatography-mass spectrometry. 3-MCPD and 1,3-DCP were analyzed using phenyl boronic acid derivatization and the liquid-liquid extraction method, respectively. The analytical method for 3-MCPD and 1,3-DCP was validated in terms of linearity, limit of detection (LOD), limit of quantitation, accuracy and precision. Consequently, the LODs of 3-MCPD and 1,3-DCP in various matrices were identified to be in the ranges of 4.18~10.56 ng/g and 1.06~3.15 ng/g, respectively.

• Journal of Pharmaceutical Investigation
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• 제37권2호
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• pp.79-84
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• 2007

This study was aimed to establish analytical method of Bi to develop a guideline of the bioequivalence test of tripotassium dicitrato bismuthate (TDB). For this purpose, a simple, specific and sensitive inductively coupled plasma-mass spectrometry (ICP/MS) method were developed and validated in human plasma. Various concentrations of bismuth standard solution (0-25ng/mL) were prepared with distilled water and human blank plasma. To 10mL of the volumetric flasks, 2mL of blank plasma was added with 8ml of distilled water. Bi standard solution was added to prepare the calibration samples and injected into ICP-MS. The plasma samples obtained from volunteers given 3 tablets of bismuth (total 900mg as TDB) were analyzed as described above. As a result, the coefficients of variation were <20% in quantitation limit (0.2 ng/mL) and <15% at the rest of concentrations. The stability test by repeated freezing-thawing cycles showed that the samples were stable only for 24hr. The stability tested for samples with a short-term period of storage at room temperature and pre-treatment prior to the analysis showed very stable over 24hr. In 8 healthy Korean subjects received Denol tablets at the dose of 900mg bismuth, AUC, $C_{max},\;T_{max}$ and half-life $(t_{1/2})$ were determined to be $198.33{\pm}173.78 ng{\cdot}hr/mL,\;64.48{\pm}27.06 ng/mL,\;0.52{\pm}0.21 hr,\;and\;5.15{\pm}2.67 hr$, respectively, from the plasma bismuth concentration-time curves. In conclusion, the method was suitable for the determination of bismuth in human plasma samples and could be applied to bioequivalence test of bismuth tablet.

• 분석과학
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• 제14권1호
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• pp.44-49
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• 2001

GC-MS와 연결된 $85{\mu}m$ 폴리 아크릴레이트 fiber의 SPME가 풍선 시료 중에 포함된 가소제를 분석하는데 사용되었다. 풍선은 IR 분광법에 의해 폴리이소프렌으로 만들어진 것으로 판명되었다. 한시간 동안 아세톤이 첨가된 물 용매에서 추출된 풍선의 가소제는 9종류의 가소제가 포함된 외부표준법에 의해 정량 하였다. 정량법은 $0.25-25{\mu}g/g$의 농도범위에서 표준 가소제들에 대하여 유효화 과정을 거쳤으며, 검출한계는 가소제에 따라 $0.11-0.38{\mu}g/g$이었고, 이러한 정량법에 의한 재현성의 RSD는 3.7-14.2%이었다. 이들 풍선 중에는 우려할만한 수준의 환경호르몬성 가소제가 검출되는 제품도 포함되어 있어 이들에 대한 규제의 필요성이 있다.

• 한국환경농학회지
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• 제25권4호
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• pp.359-364
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• 2006

Reversed-phase high-performance liquid chromatography (HPLC) techniques were developed to quantify abamectin (ABM) in paprika (Capsicum annum). Separation was achieved on a $C_{18}$ ODS column with a mobile phase of acetonitrile/water (96/4, v/v) mixture in an isocratic elution at the flow tate of 1.2 mL/min for avermectins (AVMs). The retention times were 8.0 and 9.7mins for AVM $B_{lb}$ and AVM $B_{1a}$, respectively. Residual AVMs (sum of AVM $B_{1a}$, AVM $B_{1b}$ and 8,9-Z-AVM $B_{1a}$) in the vegetable were extracted with acetonitrile, and the silica solid-phase extraction cartridges were used to purify the extract. AVMs were derivatized using trifluoroacetic acid and 1-methylimidazole, and the derivatives were determined with a fluorescence detector (excitation at 365 nm and emission at 470 nm). High and consistent recoveries, ranging from 93% to 115%, were obtained for AVM $B_{1a}$ and 8, 9-Z-AVM $B_{1a}$ at fortified levels of $20{\mu}g/kg\;and\;200{\mu}g/kg$ for paprika. The limit of quantitation (LOQ) was $2{\mu}g/kg$. The residual levels of AVMs in paprika in a field experiment from one day to seven days after the last application decreased from 18.40 to $7.59{\mu}g/kg$. The half-life $(T_{1/2})$ of AVMs in paprika was 1.47 days.

• 대한한방종양학회지
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• 제2권1호
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• pp.1-23
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• 1996

In order to research the effects of Shipjundaebotang(SDT) and Orostachys Herba(OH) on anti-tumor and immune response, the author performed this experimental study. Experimental groups are divided into five groups, which are solid extract of Orostachys Herba by water(OHW), solid extract of Orostachys Herba by ethanol(OHE), solid extract of Shipjundaebotang (SDT), solid extract of Shipjundaebotang added by solid extract of Orostachys Herba by water(SDT+OHW) and solid extract of Shipjundaebotang added by solid extract of Orostachys Herba by ethanol (SDT+OHE). In these experimental studies, extension of survival days for anti-tumor effect was observed, and de layed type hypersensitivity and rosette forming cell for cell-mediated immune response, hemagglutinin titers and hemolysin titers for humeral immune response, spleenic natural killer cell activity and carbon clearance (K-index) in vitro were measured with mice. The result were summerized as follows: I. SDT, SDT+OHW and SDT+OHE treated groups were significantly recognized to extend the survival days of tumor bearing mice as compared with the control group. 2. Delayed type hypersensitivity was significantly increased in SDT, SDT+OHW and SDT+OHE treated groups as compared with control group. 3. Hemagglutinin titer was increasred in all sample groups as compared with control group, but not significantly. 4. Hemolysin titers was significantly increased in SDT, SDT+OHW and SDT+OHE treated groups as compared with control group, and SDT+OHE treated group showed the increasing effect with significance as compared with the other sample groups. 5. For the effect of roselle forming cell quantitation, SDT, SDT+OHW and SDT+OHE treated groups showed the increasing effect with significance as compared with control group. 6. Natural killer cell activity was significantly increased in SDT+OHW as compared with comrol group, but the other groups, except OHW and SDT+OHW treated groups, revealed the increasing effect as compared with control group, but the significance was not admitted. 7. For the effect of K-index(Carbon clearance), SDT, SDT+OHW and SDT+OHE treated groups showed the increas ing effect with significance as compared with control group. 8. The study didn't show that Orostachys herba had any significance with survival days, anti-tumor effect and immune re sponse.

• Journal of Pharmaceutical Investigation
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• 제35권4호
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• pp.287-293
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• 2005

The purpose of the present study was to evaluate the bioequivalence of two glimepiride tablets, Amaryl tablet (Handok & Aventis Korea, reference drug) and Mepiril tablet (Myungmoon Pharm. Co., Ltd., Korea, test drug), according to the guidelines of Korea Food and Drug Administration (KFDA). After adding an internal standard (glibenclamide) to human plasma, plasma samples were extracted using 1mL of methyl tertiary butyl ether. Compounds extracted were analyzed by reverse-phase HPLC with multiple reaction monitoring (MRM) mode analyte detection. This method for determination glimepiride proved accurate and reproducible, with a limit of quantitation of 2 ng/mL in human plasma. Twenty-four healthy male Korean volunteers received each medicine at the glimepiride dose of 2 mg in a $2{\times}2$ crossover study. There was a one-week washout period between the doses. Plasma concentrations of glimepiride were monitored by a LC-MS/MS for over a period of 12 hr after the administration. $AUC_t$ (the area under the plasma concentration-time curve from time zero to 12 hr) was calculated by the linear trapezoidal rule method. $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the plasma concentration-time data. Analysis of variance was carried out using logarithmically transformed $AUC_t$ and $C_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the crossover design was properly performed. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for Amaryl/Mepiril were log 0.9583-log 1.1357 and log 1.0570-log 1.2376, respectively. These values were within the acceptable bioequivalence intervals of log 0.80-log 1.25. Taken together, our study demonstrated the bioequivalence of Amaryl and Mepiril with respect to the rate and extent of absorption.

• Journal of Pharmaceutical Investigation
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• 제35권6호
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• pp.431-436
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• 2005

A high-performance liquid chromatographic method was validated for quantitation of nadolol in human plasma. Nadolol and internal standard, pindolol, were extracted with tert-butyl methyl ether after addition of 10 M sodium hydroxide solution. The analytes were separated on a reverse phased C18 column using a mobile phase consisting of 0.05 M ammonium phosphate monobasic buffer, acetonitrile and methanol (81: 17:2 v/v/v) and detected using a fluorescence detector (excitation wavelength 230 nm, emission wavelength 294 nm). The method was specific and sensitive enough to detect as low as 3 ng/mL of nadolol in human plasma. Linear calibration range was 3-150 ng/mL with correlation coefficient greater than 0.999. The overall accuracy was in the range of 96.8 to 103% and precision C.V.(%) 7.30 to 12.2%. The recovery was approximately 100% and stability was confirmed during storage and sample preparation. The present HPLC method was successfully applied to study bioavailability after oral administration of 80 mg of nadolol in healthy Korean subjects. The mean $AUC_{t}$ was $1968{\pm}397\;ng{\cdot}hr/mL$ and $C_{max}$ of $186{\pm}79.3\;ng/mL$ was reached at $3.5{\pm}0.76\;hr$. The mean $t_{1/2}$ of nadolol was $17.3{\pm}2.59\;hr$.

• 분석과학
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• 제14권5호
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• pp.410-415
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• 2001

철강 시료 중의 납 분석을 위하여 수소화물 발생법을 플라스마에 납을 선택적으로 주입하고 유도결합플라스마 질량분석법으로 측정하는 분석법을 개발하였다. 납 수소화물 $PbH_4$의 생성을 위해서는 $NaBH_4$와 반응 전에 먼저 준안정 상태의 Pb(IV)로 만들어주기 위하여 산화제가 필요하다. $1000{\mu}g/mL$ 이상의 철 매질을 함유하는 시료용액으로 납 수소화물을 발생시키기 위한 최적조건을 찾는 연구를 수행하였다. 철 매질이 $10{\mu}g/mL$ 이상 존재할 때는 $K_2Cr_2O_7$이 효과적인 산화제로 작용하였으며 젖산을 가하여 감도를 향상시켰다. 시료용액의 산농도 그리고 산화제와 젖산 농도의 최적값은 철 매질의 농도에 따라 달랐다. 동위원소 희석법을 사용하여 납을 정량하였으며 철강 표준물질 NIST SRM 361, 362 분석결과는 불확도 범위안에서 검정값과 잘 일치하였다.