• Title/Summary/Keyword: Basic and Analytical Science

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Development and Assessment of New RT-qPCR Assay for Detection of HIV-1 Subtypes

  • Lim, Kwanhun;Park, Min;Lee, Min Ho;Woo, Hyun Jun;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.22 no.3
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    • pp.83-97
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    • 2016
  • The measurement of viral load in HIV-1 infected patients is essential for the establishment of a therapeutic strategy. Several commercial assays have shown shortcomings in quantifying rare genotypes of HIV-1 such as minor groups of N and O. In this study, the HIV-1 RT-qPCR assay was developed. The primers and probe of HIV-1 were designed to target the pol gene and to increase the detection efficiency of various subtypes including group N and O. The HIV-1 quantitative RT-qPCR assay was assessed for its analytical performance and clinical evaluation. The LoD was determined to 33.9 IU/ml. The LoD of several subtypes including A, C, D, CRF_01AE, F, CRF_02AG, G and H, were determined to less than 40 IU/ml. The HIV-1 quantitative RT-qPCR assay was evaluated using the China National Reference Panel of HIV-1 RNA to determine the analytical performance. The results were all within the acceptable range. The clinical evaluation was performed at Hunan CDC in China. The clinical evaluation results were compared with those of the China domestic commercial kit. A significant correlation (fresh samples; $R^2=0.84$, P<0.001, frozen samples; $R^2=0.76$, P<0.001) between the two systems was observed for 64 fresh samples and 76 frozen samples with viral loads, and the Bland-Altman plot showed good agreement (98.4%, 96.1%, respectively). In conclusion, the HIV-1 quantitative RT-qPCR assay had comparable analytical performance with several commercial kits. The study provides basic data for the research of HIV-1 diagnosis and the development of P < HIV-1 molecular diagnostic assay.

Application of Malononitrile Derivatization Method for Structural Glycomics Study in Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

  • Ahn, Yeong-Hee;Yoo, Jong-Shin
    • Journal of Photoscience
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    • v.8 no.2
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    • pp.83-86
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    • 2001
  • Structural analyses of oligosaccharide-malononitrile derivatives were conducted by matrix-assisted laser desorption/ionization post-source decay (MALDI-PSD) analysis in positive ion mode. The malononitrile derivatives of oligosaccharides, which were developed for highly sensitive detection of multi-component oligosaccharides by negative ion electrospray ionization mass spectrometry (ESI MS), were detected by positive-ion MALDI with the detection limit of 2 pmol level from the crude derivatization sample. The used matrix affected drastically the analytical results of oligosaccharide-malononitrile derivative by matrix-assisted laser desoprtion/ionization mass spectrometry (MALDI MS). The malononitrile derivatization of oligosaccharide also affect the patterns of MALDI-PSD spectra and give much more structural information than the free oligosaccharide.

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A Comparison of Sonication and Microwave-assisted Extraction Method for Speciation of Arsenic in Fish Tissue, DORM-2 (어류중 비소의 종분화 분석을 위한 초음파 추출법과 마이크로파 추출법의 비교)

  • Yoon, Cheol-Ho;Park, Yong-Chul;Hong, Jong-Ki
    • Analytical Science and Technology
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    • v.16 no.2
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    • pp.134-142
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    • 2003
  • Comparison of a microwave-assisted extraction with sonication extraction was performed for arsenic speciation in fish tissue with chromatographic separation and inductively coupled plasma mass spectrometry detection. The detection limits of arsenicals with ultrasonic nebulizerand cross-flow nebulizer were shown to be similar. The arsenicals investigated were arsenobetaine (AsB), arsenite [As(III)], dimethylarsine acid (DMA), monomethylarsonic acid (MMA), arsenate [As(v)], and phenylarsonic acid (PAA). Quantitative extraction of arsenicals from dogfish muscle, DORM-2, standard reference material of NRCC (National Research Council of Canada) was achieved using 50% (v/v) methanol-water in both extraction methods. Extraction efficiency of arsenobetaine in both methods is greater than 82% with RSDs on replicates of less than 5%. The concentrations of AsB determined in extract of microwave assisted extraction and sonication methods were $14.18{\pm}0.42mg\;kg^{-1}$ and $13.54 {\pm}0.84mg\;kg^{-1}$, respectively. And the concentrations of DMA were $0.45{\pm}0.06mg\;kg^{-1}$ and $0.44{\pm}0.06mg\;kg^{-1}$, respectively.

A Study on Determination of Iodine in Serum, Fresh Milk, and Feed Additive by Inductively Coupled Plasma-Mass Spectrometry (유도결합 플라스마-질량분석법에 의한 혈청, 생우유 및 사료첨가제중 요오드의 분석에 관한 연구)

  • Lee, Won;Park, Kyung-Su;Kim, Sun-Tae;Kim, Young-Man
    • Analytical Science and Technology
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    • v.12 no.6
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    • pp.528-533
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    • 1999
  • The total iodine is determined in serum, fresh milk and feed additive by ICP-MS. The preparation involves only a 25-fold dilution with a diluent by direct nebulization in the plasma. The diluent composition is $NH_4OH$(0.5% v/v)+$CH_3OH$(5% v/v) and the abundance of iodine is measured at m/z=127. The calibration curve was linear over the ranges $0-100{\mu}g/L$ with $R^2=0.99$ for iodine. The detection limit of this method is $0.084{\mu}g/L$ for iodine. The relative error range of milk powder SRM in optimum condition is 2.30%-4.73%. The concentration ranges of iodine in the serum, fresh milk, feed additive were $12.4-40.2{\mu}g/L$, < 0.01-3.11 mg/L, < $10^{-7}-2.60g/kg$ respectively.

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Determination of hormonal active compounds in meat (육류 식품중 호르몬 성분의 분석)

  • Seo, Jungju
    • Analytical Science and Technology
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    • v.21 no.6
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    • pp.526-534
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    • 2008
  • To determine the trace level of synthetic and natural hormones in food, the improvement of official analytical method and new development of simultaneous determination of hormones were established. On the basis of developed analytical method, the background level of natural hormones and distribution of residual hormones were monitored in meat. Target hormones were six natural hormones such as estrogens ($17{\beta}$-estradiol, $17{\alpha}$-estradiol, estrone), androgens ($17{\beta}$-testosterone, $17{\alpha}$-testosterone), and gestagens (progesterone) and three synthetic hormones such as DES, zeranol, and taleranol. These hormones were analyzed by gas chromatographymass spectrometry. Newly developed multi-residue analysis method was applied for meat sample which were collected from market in the capital region and monitored the presence of residues of synthetic and natural steroid hormones. No residue of synthetic hormones were detected and endogenous level of progesterone was detected in cattle, pig and liver samples tested.

Analytical trend of perfluorinated compounds in environmental and biota samples (환경 및 생체시료 중 과불화 화합물의 분석 동향)

  • Lee, Won-Woong;Chang, Won-Hee;Pyo, Hee-Soo;Kang, Tae-Seok;Hong, Jong-Ki
    • Analytical Science and Technology
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    • v.23 no.4
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    • pp.331-346
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    • 2010
  • Perfluorinated compounds have characteristics of resistance to heat, acidic, basic conditions and also resist water, oil, grease, pollutant. Futhermore they are used by various industrial material, nowadays, they produced in large scale for indutrial and commercial areas. However, they also resist metabolizing and degrading in environmental system (plant, animal, even human body). Moreover, in animal's bodies, PFCs can be accumulated in organ (eg; liver) and lead to liver cell necrosis even oncogenesis. Perfluorinated compounds are newly registered as new persistent organic pollutants (POPs) on Stockholm convention in 2009. Therefore necessity for analytical methodology for determination of PFCs in various environmental samples is even more increased. This study discussed sample preparation and instrumental conditions for the analysis of PFCs in environmental and biota samples.

Studies on the Separation and Concentration Method of $^{90}Sr$ in the Environmental Samples (환경시료 중 $^{90}Sr$의 분석을 위한 분리농축법 연구)

  • Won, Mi Sook;Cho, Kyu Bong;Yoon, Jang Hee;Lee, Dae Won;Shim, Yoon Bo
    • Analytical Science and Technology
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    • v.14 no.1
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    • pp.8-14
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    • 2001
  • New process to determine $^{90}Sr$ in the environmental samples was established by investigating the existing methods. The environmental samples included soil, milk, seaweed, fishes and clams, pine needles, and marine sediment. Using the developed method combined with fuming nitric acid and ion exchange resin, which could be reduced the treatment step, we could be removed Ca effectively for the determination of $^{90}Sr$. The recovery yield of $^{90}Sr$ in this method was 10% higher than those of using the fuming nitric acid only. This method could be applied to all environmental samples we choose. The content of $^{90}Sr$ in soil was the highest value in the tested environmental samples.

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SEMI-ANALYTICAL SOLUTION TO A COUPLED LINEAR INCOMMENSURATE SYSTEM OF FRACTIONAL DIFFERENTIAL EQUATIONS

  • Iqbal M. Batiha;Nashat Alamarat;Shameseddin Alshorm;O. Y. Ababneh;Shaher Momani
    • Nonlinear Functional Analysis and Applications
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    • v.28 no.2
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    • pp.449-471
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    • 2023
  • In this paper, we study a linear system of homogeneous commensurate /incommensurate fractional-order differential equations by developing a new semi-analytical scheme. In particular, by decoupling the system into two fractional-order differential equations, so that the first equation of order (δ + γ), while the second equation depends on the solution for the first equation, we have solved the under consideration system, where 0 < δ, γ ≤ 1. With the help of using the Adomian decomposition method (ADM), we obtain the general solution. The efficiency of this method is verified by solving several numerical examples.

A Study of Determination of Minimum Detectable Activity at Measuring of Pu Isotopes in Environmental Samples (환경시료 중 Pu 측정에서 최소검출방사능 농도 산출에 대한 연구)

  • Lee, Myung-Ho;Cho, Young-Hyun;Choi, Gun-Sik;Lee, Chang-Woo;Shin, Hyun-Sang
    • Analytical Science and Technology
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    • v.14 no.1
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    • pp.64-71
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    • 2001
  • In this paper, the formula of lower limit of detection considering the measuring time of sample and background was derived using the basic concepts of lower limit of detection. Among parameters affecting the determination of MDA value, the MDA values were calculated with variation of amount of sample and measuring time of sample and background. The results of adequate division between the measuring time of sample and that of background studied in this research make it possible to evaluate confidence limits on the radio analytical results in the environmental sample.

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Simultaneous Separation Analysis of Some Metal Ions in Piperidinedothiocarbamate Chelates by Reversed-Phase Liquid Chromatography (역상 액체 크로마토그래피에 의한 몇가지 금속 이온들의 Piperidinedithiocarbamate 킬레이트 동시분리분석)

  • Lee, Won;Bahng, Seung-Hoon;Kim, Mi-Kyoung
    • Analytical Science and Technology
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    • v.13 no.1
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    • pp.27-33
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    • 2000
  • Simultaneous separation and analysis of Ni(II), Pd(II), Co(II), Cu(II) and Hg(II) in peperidinedithiocarbamate (PDTC) chelates were investigated by reversed phase liquid chromatography. The optimum conditions for the separation of PDTC metal chelates were examined with respect to the pH, extraction solvent, and mobile phase strength on Novapak $C_{18}$ column using methanol/water mixture as mobile phase. All metal PDTC chelates were eluted in an acceptable range of capacity factor value ($0{\leq}log\;k^{\prime}{\leq}1$). The linear calibration curves were obtained in the concentration range of $0{\sim}1.2{\mu}g/mL$ for five metal ions, and also good precision in the range of 1.96~3.41% RSD was obseved. Under the optimum conditions, trace metat ions in the composite water sample were successfully separated and determined with relative error of ${\pm}2.0%$.

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