• Clinical and Experimental Pediatrics
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• 제50권2호
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• pp.143-150
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• 2007

목 적: 인체의 항 PRP IgG 항체를 정량적으로 측정하기 위한 방법인 표준화된 효소면역법의 타당성을 연구하는 것이 목적이다. 방 법: 인체의 항 PRP IgG 항체를 정량적으로 측정하기 위한 방법인 표준화된 효소면역법의 타당성을 연구하기 위해 특이성, 반복성, 실험실내 정밀성, 정확성, 최소 정량 한계, 및 안정성을 평가하였다. 결 과: 본 연구에서 사용한 효소면역법은 검사에 사용된 항원(HbO-HA)에 특이성을 보였으며 반복성, 실험실내 정밀성 등의 정밀성은 허용기준(반복성 : $CV{\leq}15%$, 실험실내 정밀성 : $CV{\leq}20%$)을 만족하였다. 정확성은 28개 혈청을 대상으로 한 RABA 정량결과와 효소 면역법 정량결과 비교시험에서 높은 상관계수를 보였고 첨가 회복 검사 결과 허용기준($100{\pm}20%$)을 만족하였다. 최소 정량 한계 시료 정량결과의 정밀성과 정확성은 공칭 양의 -14.7~-4.7%로 모두 허용기준(정밀성 : $CV{\leq}25%$, 정확성 : ${\pm}25%$)을 만족하였다. 안정성 중 냉 해동 안정성과 단기 온도 안정성도 모두 허용기준(${\pm}20%$ 이내)을 만족하였다. 결 론: 이상의 결과로 이화여자대학교 의과학연구소 백신효능연구센터에서 시행한 본 효소 면역법은 혈액 내에 존재하는 항 PRP IgG 항체를 정량적으로 측정하는 시험법으로 적절하였다.

• Biomolecules & Therapeutics
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• 제6권3호
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• pp.289-295
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• 1998

The bioequivalence of two nilvadipine products was evaluated in 16 normal male volunteers (age 22-32 yr, body weight 57-80 kg) following sidle oral dose. Test product was Overca $l_{R}$ tablet (Choong-Wae Pharm. Corp., Korea) and reference product was Nivadi $l_{R}$ tablet (Hyundai Pharm. Corp., Korea). Both products contain 4 mg of nilvadipine. One tablet of the test or the reference product was administered to the volunteers, respectively, by randomized two period cross-over study (2$\times$2 Latin square method). The determination of nilvadipine was accomplished using a validated capillary column GC with electron-capture detection. As a result of the assay validation, the quantiflcation of nilvadipine in human plasma by this technique was possible down to 0.5 ng/ml using 1 ml of plasma. Absolute overall recovery from five replicate analyses of nilvadipine-spiked sample were 88.4$\pm$ 10.24% (mean$\pm$ 5.D.) for human plasma of 10 ng/ml. The coefficients of variation (C.V.) were less than 20% and the actual concentration of nilvadipine measured by GC ranged from 80 to 99% in all plasma. Average drug concentrations at each sampling time and pharmacokinetic parameters calculated were not significantly different between two products (p>0.05); the area under the curve from time zero to 8 hr (AUCo-$_{8 hr}$) (22.8$\pm$5.90 vs 22.2$\pm$6.10 ng . hr/ml), maximum plasma concentration ( $C_{max}$) (10.0$\pm$2.85 vs 9.3$\pm$3.28 ng/ml) and time to reach maximum plasma concentration ( $T_{max}$) (1.2$\pm$0.31 vs 1.3 $\pm$0.47 hr). The differences of mean AU $Co_{8hr}$ $C_{max}$, and $T_{max}$ between the two products (2.25, 7.65, and 10.30%, respectively) were less than 20%. The power (1-$\beta$) and treaeent difference (7) for AU $Co_{8hr}$, and $C_{max}$ were more than 0.8 and less than 0.2, respectively. Although the power for Tmax was under 0.8, Tm\ulcorner of the two products was not significantly different from each other (p>0. 05). These results suggest that the bioavailability of Overeat tablet is not significantly different from that of Nivadil tablet. Therefore, two products are bioequivalent based on the current results.sults.lts.lts.lts.

• 한국식품영양과학회지
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• 제41권12호
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• pp.1753-1757
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• 2012

LC-MS/MS를 사용함으로써 인체 혈장에서 ginsenoside Rb1의 분석법을 개발하고 검증하였다. 유속 0.9 mL/min에 이동상 0.1% formic acid가 첨가된 water와 methanol을 사용하여 기울기 용리 조건으로 설정하였으며 사용한 분석 칼럼은 C 18($4.6mm{\times}150mm$, particle size 5 ${\mu}m$)을 사용하여 분리하였다. MRM(multiple reaction monitoring) 방법의 전기 분무 이온화 이온 분석기로 모니터링 하여 분석하였다. 인체 혈장 샘플은 액체-액체 추출방법에 의해 acetone과 water가 섞인 용액으로 추출하였다. 이 분석의 검량선 범위는 10~500 ng/mL이며 상관계수는 0.9995를 나타냈다. 일내, 일간의 정밀성 농도범위는 상관계수 5.8% 그리고 정확성은 96.0~104.6%로 나타났다. 이 LC-MS/MS를 이용한 인체 혈장의 ginsenoside Rb1의 연구가 약동학 연구에 적용할 수 있을 거라 생각한다.

• 대한임상검사과학회지
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• 제38권2호
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• pp.117-124
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• 2006

The analytical measurement range (AMR) is the range of analyte values that a method can directly measure on a specimen without any dilution, concentration, or other pretreatment not part of the usual assay process. The linearity of the AMR is its ability to obtain test results which are directly proportional to the concentration of analyte in the sample from the upper and lower limit of the AMR. The AMR validation is the process of confirming that the assay system will correctly recover the concentration or activity of the analyte over the AMR. The test specimen must have analyte values which, at a minimum, are near the low, midpoint, and high values of the AMR. The AMR must be revalidated at least every six months, at changes in major system components, and when a complete change in reagents for a procesure is introduced; unless the laboratory can demonstrate that changing the reagent lot number does not affect the range used to report patient test results. The AMR linearity was total protein (0-16.6), albumin (0-8.1), total bilirubin (0-18.1), alkaline phosphatase (0-1244.3), aspartate aminotransferase (0-1527.9), alanine aminotransferase (0-1107.9), gamma glutamyl transpeptidase (0-1527.7), creatine kinase (0-1666.6), lactate dehydrogenase (0-1342), high density lipoprotein cholesterol (0.3-154.3), sodium (35.4-309), creatinine (0-19.2), blood urea nitrogen (0.5-206.2), uric acid (0-23.9), total cholesterol (-0.3-510), triglycerides (0.7-539.6), glucose (0-672.7), amylase (0-1595.3), calcium (0-23.9), inorganic phosphorus (0.03-17.0), potassium (0.1-116.5), chloride (3.3-278.7). We are sure that materials for the AMR affect the evaluation of the upper limit of the AMR in the process system.

• Journal of Pharmaceutical Investigation
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• 제35권6호
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• pp.461-465
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• 2005

A simple HPLC method with ultraviolet detection of nicardipine in human plasma was developed and validated. After drug extraction with solid phase extraction (SPE) method, chromatographic separation of nicardipine in plasma was achieved at $30^{\circ}C$ with a $C_{18}$ column and acetonitrile-0.02% phosphate buffer mixture (with 0.02% triethylamine, final pH 7.0), as mobile phase. Quantitative determination was performed by ultraviolet detection at 254 nm. The method was specific and validated with a limit of quantification of 5 ng/mL. The intra- and inter-day precision and accuracy were acceptable for all quality control samples including the lower limit of quantification. The applicability of the method was demonstrated by analysis of plasma after oral administration of a single 40 mg dose to 8 healthy subjects. From the plasma nicardipine concentration versus time curves, the mean $AUC_{t}$, was $134.04{\pm}59.72\;ng\;hr/mL$ and $C_{max}$ of $108.65{\pm}69.17\;ng/mL$ reached 1.5 hr after administration. The mean biological half-life of nicardipine was $3.93{\pm}0.82\;hr$. Based on the results, this simple and validated assay method could readily be used in any pharmacokinetic or bioequivalence studies using human.

• 한국방사성폐기물학회:학술대회논문집
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• 한국방사성폐기물학회 2003년도 가을 학술논문집
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• pp.534-538
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• 2003

Final disposal of radioactive waste generated from Nuclear Power Plant (NPP) requires the detailed knowledge of the natures and quantities of radionuclides in waste package. Many of these radionuclides are difficult to measure and expensive to assay. Thus it is suggested to the Indirect method by which the concentrations of DTM (Difficult-to-Measure) nuclide is decided using the relation of concentrations (Scaling Factor) between Key (Easy-to-Measure) nuclide and DTM nuclide with measured concentrations of Key nuclide. In general, scaling factor is determined by using of log mean average (LMA) and regression. These methods are adequate to apply most corrosion product nuclides. But in case of fission product nuclides and some corrosion product nuclides, the predicted values aren't well matched with the original values. In this study, the models using artificial neural network (ANN) for C-14 and Sr-90 are compared with those using LMA and regression. The assessment of models is executed in the two parts divided by a training part and a validation part. For all of two nuclides in the training part, the predicted values using ANN are well matched with the measured values compared with those using LMA and regression. In the validation part, the accuracy of the predicted values using ANN is better than that using LMA and is similar to or better than that using regression. It is concluded that the predicted values using ANN model are better than those using conventional model in some nuclides and ANN model can be used as the complement of LMA and regression model.

• 한방재활의학과학회지
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• 제30권2호
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• pp.67-75
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• 2020

Objectives SJ004 is a natural herbal medicine that contains Acyranthes japonica Nakai and Eucommia ulmoides Oliver traditionally used for joint and spinal diseases. This study aimed to establish an efficient method of extracting SJ004 to standardize using the yield, high-performance liquid chromatography (HPLC), and antioxidant assay. Methods SJ004 was extracted with distilled water, 70% and 100% of ethyl alcohol (EtOH). The method validation of 20-hydroxyecdysone and pinoresinol diglucoside was determined by HPLC-photo diode array and the content of SJ004 was calculated. The antioxidant activity of each extract was compared and measured using total flavonoids, total phenolic compounds, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), and ferric reducing antioxidant power according to the standard protocol. Results The yield was highest in pure water extract and lowest in 100% EtOH. But, the content of marker compounds indicating 20-hydroxyecdysone and pinoresinol diglucoside was highest in 100% EtOH extract. In the physiological activity measurement using antioxidant activity, 100% ethanol extract was highest. The limit of detection indicating 20-hydroxyecdysone and pinoresinol diglucoside were analyzed 0.33 ㎍/mL, 0.1616 ㎍/mL, and the limit of quantification were analyzed 1.01 ㎍/mL and 0.49 ㎍/mL respectively. Conclusions The experimental results showed that the extraction conditions have a significant effect on content of marker compounds and antioxidant activity. As a result of method validation, SJ004 was standardized by 20-hydroxyecdysone and pinoresinol diglucoside.

• 분석과학
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• 제36권4호
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• pp.180-190
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• 2023

The presence of process related impurities in any drug or the drug product was associated with its safety, stability and efficacy. The overall literature survey proved that there is no method published on the assessment of process related impurities in brigatinib. In this study, a simple, reliable and stable HPLC qualitative method was reported for quantification of process related impurities with easy and quick extraction procedure. The impurities along with standard brigatinib was resolved on Lichrospher^® C18 (250 mm × 4.6 mm; 5 ㎛ particle size) column in room temperature using methanol, acetonitrile, pH 4.5 phosphate buffer in 55:25:20 (v/v) at 1.0 mL/min as mobile phase and UV detection at 261 nm. The method produces well resolved peaks at retention time of 4.60 min, 12.28 min, 3.37 min, 7.34 min and 8.39 min respectively for brigatinib, impurity A, B, C and D. The method produces a very sensitive detection limit of 0.0065 ㎍/mL, 0.0068 ㎍/mL, 0.0053 ㎍/mL and 0.0058 ㎍/mL for impurity A, B, C and D respectively with calibration curve linear in the concentration range of 22.5-135 ㎍/mL for brigatinib and 0.0225-0.135 ㎍/mL for impurities. The method produces all the validation parameters under the acceptable level and doesn't produces any considerable changes in peak area response while minor changes in the developed method conditions. The method can effectively resolve the unknown stress degradation products along with known impurities with less % degradation. The method can efficiently resolve and quantify the impurities in formulation and hence can suitable for the routine quality analysis of brigatinib in raw material and formulation.

• 대한수의학회지
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• 제44권4호
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• pp.655-662
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• 2004

The aim of this work was the validation of a rapid real-time PCR assay based on TaqMan technology for the unequivocal identification of infectious canine hepatitis (ICH) virus, to be used directly on DNA purified from blood specimens. A real-time PCR system targeting at the E3 ORFA gene sequence of canine adenovirus type 1 was optimized and validated through comparative analysis of samples using conventional PCR system. The real-time PCR assay based on TaqMan technology could disclose 23 (37.7%) out of 61 samples as PCR positive. In contrast, 18 (29.5%) samples were found PCR positive when conventional PCR was applied on these samples. The use of the ABI Prism 7700 sequence detection system allowed the efficient determination of the amplified product accumulation through a fluorogenic probe. The entire real-time TaqMan PCR assay, including DNA extraction, amplification, and detection could be completed within 3 hours. The detection method of real-time TaqMan PCR assay was 1,000 times more sensitive than conventional PCR. Real-time TaqMan probe and primer set developed and optimized in this study is a sensitive, rapid and accurate method for detection of ICH virus and can be effective screening tool for the detection of ICH in a diagnostic laboratory routines.

• Bulletin of the Korean Chemical Society
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• 제35권5호
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• pp.1279-1284
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• 2014

The binding of TATA-binding protein (TBP) to the TATA-box containing promoter region is aided by many other transcriptional factors including TFIIA and TFIIB. The mechanistic insight into the assembly of RNA polymerase II preinitation complex (PIC) has been gained by either directly altering a function of target protein or perturbing molecular interactions using drugs, RNAi, or aptamers. Aptamers have been found particularly useful for studying a role of a subset of PIC on transcription for their ability to inhibit specific molecular interactions. One major hurdle to the wide use of aptamers as specific inhibitors arises from the difficulty with traditional assays to validate and determine specificity, affinity, and binding epitopes for aptamers against targets. Here, using a technique called the bio-layer interferometry (BLI) designed for a label-free, real-time, and multiplexed detection of molecular interactions, we studied the assembly of a subset of PIC, TBP binding to TATA DNA, and two distinct classes of aptamers against TPB in regard to their ability to inhibit TBP binding to TFIIA or TATA DNA. Using BLI, we measured not only equilibrium binding constants ($K_D$), which were overall in close agreement with those obtained by electrophoretic mobility shift assay, but also kinetic constants of binding ($k_{on}$ and $k_{off}$), differentiating aptamers of comparable KDs by their difference in binding kinetics. The assay developed in this study can readily be adopted for high throughput validation of candidate aptamers for specificity, affinity, and epitopes, providing both equilibrium and kinetic information for aptamer interaction with targets.