• Title/Summary/Keyword: Quantitation

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Quantitative Determination of Ascaridole, Carvacrol and p-Cymene in the Biopesticides Products Derived from Chenopodium ambrosioides L. Extracts by Gas Chromatography

  • Yang, Ji-Yeon;Ryu, Song-Hee;Lim, Sung-Jin;Choi, Geun-Hyoung;Park, Byung-Jun
    • Korean Journal of Environmental Agriculture
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    • v.35 no.3
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    • pp.211-215
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    • 2016
  • BACKGROUND: The commercial biopesticides containing Chenopodium ambrosioides L. extracts which have been registered as one of the ingredients of the commercial biopesticide by the organic agriculture materials, and have been widely used in Republic of Korea. However, the quantitative analysis method of the active substances for the commercial biopesticides containing C. ambrosioides L. extract has not been conducted.METHODS AND RESULTS: To analyze the quantitative analysis of ascaridole, carvacrol, and p-cymene as active substances of C. ambrosioides L. extract, hydrophilic lipophilic balance cartridge was used for solid phase extraction. The active substances were analyzed by the gas chromatography with flame ionization detector. The limit of quantitation values of ascaridole, carvacrol, and p-cymene were 10, 5, and 2 mg/L, and the recovery rates were 96.3, 84.0, and 82.5% in liquid products and 98.3, 99.1, and 97.3% in solid products, respectively. The total content of ascaridole, carvacrol, and p-cymene in the commercial biopesticides was ranged from 0.08 to 12.75%.CONCLUSION: From these results, this method was suitable for the quantitative analysis of the active substances of commercial biopesticides containing C. ambrosioides L. extract.

Telomeres Distribution and Telomerase Activity During Chick Embryonic and Developmental Stages

  • Cho, E.J.;Kang, M.Y.;Jung, G.S.;Sohn, S.H.
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.111-111
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    • 2003
  • Telomeres are the end of chromosomes and consist of a tandem repeat sequence of (TTAGGG)n and associated proteins. Telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. Even though telomeres and telomerase have been studied extensively, very little is known about telomere dynamics in embryonic cells. This study was carried out to analyze the telomeres distribution and telomerase activity of chicken cells during embryonic and developmental stages. The target cells for analysing were sperms, ovulated ova, early embryonic cells and the cells from brain, heart, liver, kidney and germinal tissue in fetus. Telomeres distribution on target cells was analyzed by Q-FISH (Quantitation-Fluorescence in situ Hybridization) techniques using a chicken telomere repeat probe. Telomerase activity was performed by TRAP assay (Telomeric repeat Amplification Protocol) with target DNA. In results, the telomeres of chicken were found at the ends of all chromosomes. In addition, chicken had interstitial telomeres on chromosomes 1, 2 and 3. Telomerase activity was highly detectable in early embryonic cells, germinal tissues and kidney cells. Whereas telomerase activity was gradually down-regulated when the organs, including brain, heart, and liver, were developed from embryos. In the distribution of telomeric DNA on the embryonic and developmental stages, most of the cells was gradually decreased in telomere quantity during ontogenesis.

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A Study on Quantitation of Polychlorinated Biphenyls (PCBs) in Sediments (저니토중 Polychlorinated Biphenyls(PCBs) 정량에 관한 연구)

  • Shin, Ho-Sang;Oh-Shin, Yunsuk
    • Analytical Science and Technology
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    • v.12 no.2
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    • pp.151-158
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    • 1999
  • This study was conducted to measure the residual PCBs in sediments of 4 great rivers in Korea by gas chromatography-mass spectrometry. From the results, PCBs were positively detected in all samples. The residue levels of total PCBs in sediments in near of SS and OS were $290.87{\mu}g/kg$ and $221.11{\mu}g/kg$, respectively. The PCBs contamination may be in association with total organic compound (TOC) in sediments. Congener 66, 74, 90, 101, 105, 110, 118, 138, 149, 153, 180, 187 as IUPAC No were predominant species found in sediments and the polyCBs detected most aboundantly in sediments were tetraCBs and pentaCBs. From the study, it appears that the PCBs contamination in sediments of 4 great rivers is not serious and the analysis from enough many sites is needed for the next survey.

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Determination of terbutaline in human plasma by coupled column chromatography (커플드칼럼크로마토그래피에 의한 사람 혈장 중 테르부탈린의 정량)

  • Ko, Mi Young;Jeon, Sang-Seol;Kim, Kyeong Ho
    • Analytical Science and Technology
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    • v.28 no.2
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    • pp.125-131
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    • 2015
  • A method was developed and fully validated for the determination of terbutaline, a β2-receptor agonist, in human plasma. Plasma samples were prepared by solid-phase extraction with Sep-Pak silica, followed by high-performance liquid chromatography (HPLC). The terbutaline was pre-separated from the interfering components in plasma on a Luna C18 (2) column, and terbutaline and salbutamol as an internal standard were resolved and determined on a Luna Silica column. The two columns were connected by a switching valve equipped with silica pre-column. The pre-column was used to concentrate the terbutaline in the eluent from the C18 column before back-flushing onto the silica column with fluorescence detection at an excitation/emission wavelength of 276/306 nm. The method was shown to be specific by testing six different human plasma sources. Linearity was established for a concentration range of 0.4-20.0 ng/mL with a correlation coefficient of 0.9999. The lower limit of quantitation was 0.4 ng/mL with a precision of 10.1% as C.V.%.

Analysis of fatty acid methyl ester in bio-liquid by hollow fiber-liquid phase microextraction

  • Choi, Minseon;Lee, Soyoung;Bae, Sunyoung
    • Analytical Science and Technology
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    • v.30 no.4
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    • pp.174-181
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    • 2017
  • Bio-liquid is a liquid by-product of the hydrothermal carbonization (HTC) reaction, converting wet biomass into solid hydrochar, bio-liquid, and bio-gas. Since bio-liquid contains various compounds, it requires efficient sampling method to extract the target compounds from bio-liquid. In this research, fatty acid methyl ester (FAME) in bio-liquid was extracted based on hollow fiber supported liquid phase microextraction (HF-LPME) and determined by Gas Chromatography-Flame Ionization Detector (GC-FID) and Gas Chromatography/Mass Spectrometry (GC/MS). The well-known major components of biodiesel, including methyl myristate, palmitate, methyl palmitoleate, methyl stearate, methyl oleate, and methyl linoleate had been selected as standard materials for FAME analysis using HF-LPME. Physicochemical properties of bio-liquid was measured that the acidity was 3.30 (${\pm}0.01$) and the moisture content was 100.84 (${\pm}3.02$)%. The optimization of HF-LPME method had been investigated by varying the experimental parameters such as extraction solvent, extraction time, stirring speed, and the length of HF at the fixed concentration of NaCl salt. As a result, optimal conditions of HF-LPME for FAMEs were; n-octanol for extraction solvent, 30 min for extraction time, 1200 rpm for stirring speed, 20 mm for the HF length, and 0.5 w/v% for the concentration of NaCl. Validation of HF-LPME was performed with limit of detection (LOD), limit of quantitation (LOQ), dynamic range, reproducibility, and recovery. The results obtained from this study indicated that HF-LPME was suitable for the preconcentration method and the quantitative analysis to characterize FAMEs in bio-liquid generated from food waste via HTC reaction.

Development of Rapid, Safe Analytical Techniques of Aflatoxins and Their Current Regulation (Aflacxin에 대한 최신 분석법과 규제동향)

  • 정덕화
    • Journal of Food Hygiene and Safety
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    • v.5 no.3
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    • pp.131-138
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    • 1990
  • Aflatoxins is a chemically diverse group of toxic secondary metabolites that are produced by fungi and often occur in agricultural commodities. Because of their wide range of toxic effects, Aflatoxins cause severe economic losses to farmers and livestock producers and pose a health to human consuming contaminated foods. Long term prospects for biotechnological control of Aflatoxins require elucidation of the specific steps and regulation of their biosynthetic pathways . Aflatoxin determinations can be approached many ways. It is essential to safely handle all experimental materials associated with aflatoxin analysis or aflatoxigenic fungi Visual screening of suspect samples, base on the presence of conidial head of the aspergillus flavus group, and screening samples for the presence of bright greenish yellow flourescence are not chemical tests and such screening techniques may allow aflactoxin contaminated lots into commerce. Microcolumn screening procedures should always be used in conjunction with a quantitative method. Several thin layer chromatography(TLC) and high performance liquid chromatography(HPLC) methods are suitable for quantitation and are in general use. Immunochemical Methods such as the ELISA or affinity column chromatography methods are being rapidly developed. The chemical and immunochemical methods can be reliable if care is taken, using suitable controls and personnel that are well trained . All analytical laboratories should stress safety and include suitable analytical validation procedure. Especially a worldwide enquiry was undertaken in recent to obtain up-to-date information about aflatoxin legislation in as many countries of the world as possible. The information concerns aflatoxin in foodstuffs. aflatoxin MI in dairy products, aflatoxins in animal feedstuffs. Limits and regulations for aflatoxin have been expended in recent with more countries having legislation on subject, more products, and more aflatoxins covered by this legislation.

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Radionuclide Quantitation of Left-to-Right Cardiac Shunts Using Deconvolution Analysis (Deconvolution 분석(分析)을 이용(利用)한 좌우단락량(左右短絡量) 측정(測定)의 의의(意義))

  • Oh, Yeon-Sang;Lee, Myung-Chul;Cho, Bo-Youn;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.18 no.1
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    • pp.9-17
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    • 1984
  • The purpose of this study is to investigate significance of new method for quantification of left to right cardiac shunts using deconvolution analysis as an adjunct to gamma variate analysis in patients with bad bolus injection. In the present study, quantitative radionuclide angiocardiography (QRAC) was performed with and without deconvolution analysis (DA) in 37 patients with left to right shunt and 103 control patients without shunt. The results obtained were as follows; 1) The mean value of $Q_p/Q_s$ in 103 control patients was $1.10{\pm}0.12$ without DA, and was $1.01{\pm}0.03$ with DA. 2) Correlation(r) between oximetry and QRAC with DA was 0.87 and correlation(r) between oximetry and QRAC without DA was 0.61. 3) The 13 patients with left to right shunt, whose $Q_p/Q_s$ was greater than 3.0 by QRAC without DA, was studied by DA. Then the $Q_p/Q_s$ values were measurable in these 13 patients by DA and they showed significant correlation with oximetry. (r=0.68) The results indicate that deconvolution analysis technique for quantification of a left to right shunt provides more reliable and accurate shunt quantification, and reduces the influences of poor bolus injection.

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Increased glucose metabolism and alpha-glucosidase inhibition in Cordyceps militaris water extract-treated HepG2 cells

  • Kim, Dae Jung;Kang, Yun Hwan;Kim, Kyoung Kon;Kim, Tae Woo;Park, Jae Bong;Choe, Myeon
    • Nutrition Research and Practice
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    • v.11 no.3
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    • pp.180-189
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    • 2017
  • BACKGROUND/OBJECTIVES: Recent living condition improvements, changes in dietary habits, and reductions in physical activity are contributing to an increase in metabolic syndrome symptoms including diabetes and obesity. Through such societal developments, humankind is continuously exposed to metabolic diseases such as diabetes, and the number of the victims is increasing. This study investigated Cordyceps militaris water extract (CMW)-induced glucose uptake in HepG2 cells and the effect of CMW treatment on glucose metabolism. MATERIALS/METHODS: Colorimetric assay kits were used to determine the glucokinase (GK) and pyruvate dehydrogenase (PDH) activities, glucose uptake, and glycogen content. Either RT-PCR or western blot analysis was performed for quantitation of glucose transporter 2 (GLUT2), hepatocyte nuclear factor 1 alpha ($HNF-1{\alpha}$), phosphatidylinositol 3-kinase (PI3k), protein kinase B (Akt), phosphorylated AMP-activated protein kinase (pAMPK), phosphoenolpyruvate carboxykinase, GK, PDH, and glycogen synthase kinase 3 beta ($GSK-3{\beta}$) expression levels. The ${\alpha}-glucosidase$ inhibitory activities of acarbose and CMW were evaluated by absorbance measurement. RESULTS: CMW induced glucose uptake in HepG2 cells by increasing GLUT2 through $HNF-1{\alpha}$ expression stimulation. Glucose in the cells increased the CMW-induced phosphorylation of AMPK. In turn, glycolysis was stimulated, and glyconeogenesis was inhibited. Furthermore, by studying the mechanism of action of PI3k, Akt, and $GSK-3{\beta}$, and measuring glycogen content, the study confirmed that the glucose was stored in the liver as glycogen. Finally, CMW resulted in a higher level of ${\alpha}-glucosidase$ inhibitory activity than that from acarbose. CONCLUSION: CMW induced the uptake of glucose into HepG2 cells, as well, it induced metabolism of the absorbed glucose. It is concluded that CMW is a candidate or potential use in diabetes prevention and treatment.

Cholesterol Biosynthesis from Lanosterol: Development of a Novel Assay Method, Characterization, and Solubilization of Rat Hepatic Microsomal Sterol Δ7-Reductase

  • Lee, Joon-No;Paik, Young-Ki
    • BMB Reports
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    • v.30 no.5
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    • pp.370-377
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    • 1997
  • A novel assay method is described for rapid quantitation of reaction rate of sterol ${\Delta}^7$-reductase (${\Delta}^7$-SR) which catalyzes reduction of the ${\Delta}^7$-double bond of sterols. Of six different organ tissues-liver, small intestine, brain, lung, kidney, and testis-. ${\Delta}^7$-SR activity was detected only in liver (2.30 nmol/min/mg protein) and testis (0.11 nmol/min/mg protein). Using a newly developed method which employs diet-induced enzyme proteins and ergosterol as substrate, we assessed both kinetics ($K_m=210\;{\mu}M$, $V_{max}=1.93\;nmol/min/mg$) and inhibition of the rat hepatic ${\Delta}^7$-SR against well-studied cholesterol lowering agents such as triparanol ($IC_{50}=16\;{\mu}M$). 3-$\beta$-[2-(diethylamino)ethoxy]androst-5-en-17-one (U18666A) ($IC_{50}=5.2\;{\mu}M$), and trans-1.4-bis(2-chlorobenzylaminomethyl)cyclohexane dihydrochloride (AY-9944) ($IC_{50}=0.25\;{\mu}M$). Of the three well-known AY-9944-sensitive cholesterogenic enzymes (i.e., ${\Delta}^7$-SR, sterol ${\Delta}^8$-isomerase, and sterol ${\Delta}^14$-reductase). ${\Delta}^7$-SR was found to be the most sensitive enzyme with a noncompetitive inhibition of this compound ($K_i=0.109\;{\mu}M$). Substrate specificity studies of the microsomal ${\Delta}^7$-SR indicate that the relative reaction rate for 7-dehydrocholesterol and ergosterol are 5.6-fold and 1.6-fold higher than that for lathosterol. ${\Delta}^7$-SR activity was also modulated by feeding rats a diet supplemented with 0.5% ergosterol (>2.6-fold) in addition to 5.0% cholestyramine plus 0.1% lovastatin ($\simeq$5.0-fold). Finally, microsomal ${\Delta}^7$-SR was solubilized by 1.5% 3-[3-(cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) and enriched on PEG (0~10%) precipitation, which should be suitable for further purification of the enzyme.

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Quantitation of Plasma Apolipoprotein A-I with a Sandwich Type Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibodies

  • Lee, Min-Gyu;Kang, Jae-Seon;Jeong, Jae-Yeon;Jue, Dae-Myung;Kim, Hack-Joo
    • BMB Reports
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    • v.30 no.6
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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.

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