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Characterization of degradation products of the Balsalazide by Mass spectrometry: Optimization of stability-indicating HPLC method for separation and quantification of process related impurities of Balsalazide

  • Chilakabattina Naga Narasimha Babu;Ch. Srinivasa Reddy;Bhagya Kumar Tatavarti;M. Radha Madhavi;Venkateswara Rao Anna
    • Analytical Science and Technology
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    • v.37 no.1
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    • pp.25-38
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    • 2024
  • The study aimed to investigate a novel approach by utilizing liquid chromatography (LC) and liquid chromatography-mass spectrometry (LC-MS) to separate, identify and characterize very nominal quantities of degradation products (DPs) of balsalazide along with its process related impurities without isolation from their reaction mixtures. The impurities along with balsalazide were resolved on spherisorb ODS2 (250×4.6 mm, 5.0 ㎛) column at room temperature using 0.2 M sodium acetate solution at pH 4.5 and methanol in the ratio of 55:45 (v/v) as mobile phase pumped isocratically at 1.0 mL/min as mobile phase and UV detection at 255 nm. The method shows sensitive detection limit of 0.003 ㎍/mL, 0.015 ㎍/mL and 0.009 ㎍/mL respectively for impurity 1, 2 and 3 with calibration curve liner in the range of 50-300 ㎍/mL for balsalazide and 0.05-0.30 for its impurities. The balsalazide pure compound was subjected to stress studies and a total of four degradation products (DPs) were formed during the stress study and all the DPs were characterized with the help of their fragmentation pattern and the masses obtained upon LC-MS/MS. The DPs were identified as 3-({4-[(E)-(4-hydroxyphenyl) diazenyl]benzoyl}amino)propanoic acid (DP 1), 4-[(E)-(4-hydroxyphenyl)diazenyl] benzamide (DP 2), 5-[(E)-(4-carbamoylphenyl)diazenyl]-2-hydroxybenzoic acid (DP 3) and 3-({4-[(E)-phenyldiazenyl]benzoyl}amino)propanoic acid (DP 4). Based on findings, it was concluded that, the proposed method was successfully applicable for routine analysis of balsalazide and its process related impurities in pure drug and formulations and also applicable for identification of known and unknown impurities of balsalazide.

Study on Relaxing Effect of Oxybutynin on the Contractile Response of Arterial Smooth Muscle (동맥근 수축에 대학 Oxybutynin의 이완효과에 관한 연구)

  • Ko, Jae-Ki;Baik, Yung-Hong
    • The Korean Journal of Pharmacology
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    • v.24 no.1
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    • pp.93-101
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    • 1988
  • Pharmacological actions of an antispasmodic agent, oxybutynin were investigated in the isolated procine coronary arteries. The coronary rings were contracted by acetylcholine (ACh) and KCl in a dose-dependent fashion. The ACh-induced contractions were signifcantly potentiated by removal of endothelium and $EC_{50}=0.52\;{\mu}M$ of intact endothelial rings was about 2 times greater than $EC_{50}=0.28\;{\mu}M$ of rings without the endothelium. These results suggest that the endothelium plays an inhibitory role in ACh-induced contraction. Oxybutynin and atropine inhibited dose-dependently $1.0\;{\mu}M$ ACh-induced contraction and atropine inhibited dose-dependently $1.0\;{\mu}M$ ACh-induced contraction and the $IC_{50s}$ were 11.0 nM and 0.47 nM, respectively. Atropine did not affect 35 mM KCl-induced contraction but oxybutynin inhibited the contraction to the basal tension in a dose-dependent manner. The $IC_{50}$ of oxybutynin on the KCl-induced contraction was $49.7\;{\mu}M$. The dose-response curve to ACh was parallelly shifted to the right by pretreating coronary rings with $IC_{50}$ of atropine (0.47 nM) or oxybutynin (11.0 nM) but the curve to KC1 was rightward shifted in a noncompetitive manner under pretreatment with $IC_{50}$ of oxybutynin $(49.7\;{\mu}M$). Oxybutynin inhibited $0.1\;{\mu}M$ Bay K 8644-induced contraction to the basal tension in a dose dependent manner, but $35\;{\mu}M$ histamine-induced contraction was inhibited to only 50e/e of the original level even in maximal concentration $(5{\times}10^{-4}M)$ of oxybutynin. These results suggest that oxybutynin causes antispasmodic action through sensitive blocking action on muscarinic receptors and inhibitory action on calcium influx in the procine coronary artery.

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BETTI NUMBERS OF GAUSSIAN FIELDS

  • Park, Changbom;Pranav, Pratyush;Chingangbam, Pravabati;Van De Weygaert, Rien;Jones, Bernard;Vegter, Gert;Kim, Inkang;Hidding, Johan;Hellwing, Wojciech A.
    • Journal of The Korean Astronomical Society
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    • v.46 no.3
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    • pp.125-131
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    • 2013
  • We present the relation between the genus in cosmology and the Betti numbers for excursion sets of three- and two-dimensional smooth Gaussian random fields, and numerically investigate the Betti numbers as a function of threshold level. Betti numbers are topological invariants of figures that can be used to distinguish topological spaces. In the case of the excursion sets of a three-dimensional field there are three possibly non-zero Betti numbers; ${\beta}_0$ is the number of connected regions, ${\beta}_1$ is the number of circular holes (i.e., complement of solid tori), and ${\beta}_2$ is the number of three-dimensional voids (i.e., complement of three-dimensional excursion regions). Their sum with alternating signs is the genus of the surface of excursion regions. It is found that each Betti number has a dominant contribution to the genus in a specific threshold range. ${\beta}_0$ dominates the high-threshold part of the genus curve measuring the abundance of high density regions (clusters). ${\beta}_1$ dominates the genus near the median thresholds which measures the topology of negatively curved iso-density surfaces, and ${\beta}_2$ corresponds to the low-threshold part measuring the void abundance. We average the Betti number curves (the Betti numbers as a function of the threshold level) over many realizations of Gaussian fields and find that both the amplitude and shape of the Betti number curves depend on the slope of the power spectrum n in such a way that their shape becomes broader and their amplitude drops less steeply than the genus as n decreases. This behaviour contrasts with the fact that the shape of the genus curve is fixed for all Gaussian fields regardless of the power spectrum. Even though the Gaussian Betti number curves should be calculated for each given power spectrum, we propose to use the Betti numbers for better specification of the topology of large scale structures in the universe.

Primary somatosensory cortex and periaqueductal gray functional connectivity as a marker of the dysfunction of the descending pain modulatory system in fibromyalgia

  • Matheus Soldatelli;Alvaro de Oliveira Franco;Felipe Picon;Juliana Avila Duarte;Ricardo Scherer;Janete Bandeira;Maxciel Zortea;Iraci Lucena da Silva Torres;Felipe Fregni;Wolnei Caumo
    • The Korean Journal of Pain
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    • v.36 no.1
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    • pp.113-127
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    • 2023
  • Background: Resting-state functional connectivity (rs-FC) may aid in understanding the link between painmodulating brain regions and the descending pain modulatory system (DPMS) in fibromyalgia (FM). This study investigated whether the differences in rs-FC of the primary somatosensory cortex in responders and non-responders to the conditioned pain modulation test (CPM-test) are related to pain, sleep quality, central sensitization, and the impact of FM on quality of life. Methods: This cross-sectional study included 33 females with FM. rs-FC was assessed by functional magnetic resonance imaging. Change in the numerical pain scale during the CPM-test assessed the DPMS function. Subjects were classified either as non-responders (i.e., DPMS dysfunction, n = 13) or responders (n = 20) to CPM-test. A generalized linear model (GLM) and a receiver operating characteristic (ROC) curve analysis were performed to check the accuracy of the rs-FC to differentiate each group. Results: Non-responders showed a decreased rs-FC between the left somatosensory cortex (S1) and the periaqueductal gray (PAG) (P < 0.001). The GLM analysis revealed that the S1-PAG rs-FC in the left-brain hemisphere was positively correlated with a central sensitization symptom and negatively correlated with sleep quality and pain scores. ROC curve analysis showed that left S1-PAG rs-FC offers a sensitivity and specificity of 85% or higher (area under the curve, 0.78, 95% confidence interval, 0.63-0.94) to discriminate who does/does not respond to the CPM-test. Conclusions: These results support using the rs-FC patterns in the left S1-PAG as a marker for predicting CPM-test response, which may aid in treatment individualization in FM patients.

2-Parameter High Frequency Combustion Instability Model (2-파라메타 모델에 의한 고주파 연소불안정 해석)

  • 조용호;윤웅섭
    • Journal of the Korean Society of Propulsion Engineers
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    • v.1 no.2
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    • pp.74-83
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    • 1997
  • The definition of burning admittance and conventional n-$\tau$ stability rating technique are combined to investigate the high frequency combustion instabilities inside the cylindrical combustion chamber. Perturbed flow variables are written as the sum of fluctuating and time-averaged mean quantities on the assumption that the terms of the order higher than unity are sufficiently small, hence linearized governing equations could be formulated. Chamber admittances up and downstream of the flame front calculated with appropriate boundary conditions result in the burning admittance and corresponding n-$\tau$ neutral stability curve. Configurational and operational design factors are tested to detect the unstable wave-induced LOX-RP1 combustion instabilities. Operational design factors, e.g. pressure or O/F ratio, appear less influential to drive high frequency instability while the location of the flame front and configurational factors enhance or deteriorate the stabilities strongly. Conclusively, LOX-RP1 combustion inside the cylindrical combustion chamber is apt to be unstable against long residence time and shortened chamber length.

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Estimation of Estrogenic Activity by E-screen Assay and Stepwise Analysis of Endocrine Disruptors in Pilot Sewage Treatment Plant (파일럿 규모의 하수 처리 공정별 E-screen Assay에 의한 에스트로겐 활성과 내분비계 장애물질의 농도 평가)

  • Lee, Ji-Ho;Park, Jong-Yol;Ra, Jin-Sung;Duong, Cuong N.;Lee, Byoung-Cheun;Kim, Sang-Don
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.7
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    • pp.697-703
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    • 2006
  • Endocrine disruptors were measured with GC/MS in effluents discharged from sewage treatment processes in pilot scale for the purpose of water reuse. From that analysis, we compared the removal rate of them by treatment processes. Nonylphenol was mainly detected in effluents and high concentration from 0.36 to 0.94 ${\mu}g/L$. $17{\beta}$-estradiol(E2) and $17{\alpha}$-ethynylestradiol(EE2) were detected as below the limit of detection in effluent. Endocrine disruptors were removed effectively in the range from 50 to 100% by treatment process. EC50 value($9.0{\times}10^{-3}$ M) of $17{\beta}$-estradiol(E2) by dose response curve of E-screen assay has higher than that of bisphenol A($2.736{\times}10^{-5}M$) and p-octylphenol($9.760{\times}10^{-6}$ M). These results showed that alkylphenols have lower relative estrogen potency than other estrogens such as $17{\beta}$-estradiol(E2). Calculated estrogenic activity(ng-EEQ/L) was 2 times higher than measured total estrogenic activity which estimated by E-screen assay. Moreover estrogenic activity of effluent by treatment process showed very low as below 1 ng-EEQ/L.

Growth and Electrical Properties of ZnAl2Se4 Single Crystal Thin Film by Hot Wall Epitaxy (Hot Wall Epitaxy(HWE)법에 의한 ZnAl2Se4 단결정 박막 성장과 가전자대 갈라짐에 대한 광전류 연구)

  • Park, Hyangsook;Bang, Jinju;Lee, Kijung;Kang, Jongwuk;Hong, Kwangjoon
    • Korean Journal of Materials Research
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    • v.23 no.12
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    • pp.714-721
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    • 2013
  • A stoichiometric mixture of evaporating materials for $ZnAl_2Se_4$ single-crystal thin films was prepared in a horizontal electric furnace. These $ZnAl_2Se_4$ polycrystals had a defect chalcopyrite structure, and its lattice constants were $a_0=5.5563{\AA}$ and $c_0=10.8897{\AA}$.To obtain a single-crystal thin film, mixed $ZnAl_2Se_4$ crystal was deposited on the thoroughly etched semi-insulating GaAs(100) substrate by a hot wall epitaxy (HWE) system. The source and the substrate temperatures were $620^{\circ}C$ and $400^{\circ}C$, respectively. The crystalline structure of the single-crystal thin film was investigated by using a double crystal X-ray rocking curve and X-ray diffraction ${\omega}-2{\theta}$ scans. The carrier density and mobility of the $ZnAl_2Se_4$ single-crystal thin film were $8.23{\times}10^{16}cm^{-3}$ and $287m^2/vs$ at 293 K, respectively. To identify the band gap energy, the optical absorption spectra of the $ZnAl_2Se_4$ single-crystal thin film was investigated in the temperature region of 10-293 K. The temperature dependence of the direct optical energy gap is well presented by Varshni's relation: $E_g(T)=E_g(0)-({\alpha}T^2/T+{\beta})$. The constants of Varshni's equation had the values of $E_g(0)=3.5269eV$, ${\alpha}=2.03{\times}10^{-3}eV/K$ and ${\beta}=501.9K$ for the $ZnAl_2Se_4$ single-crystal thin film. The crystal field and the spin-orbit splitting energies for the valence band of the $ZnAl_2Se_4$ were estimated to be 109.5 meV and 124.6 meV, respectively, by means of the photocurrent spectra and the Hopfield quasicubic model. These results indicate that splitting of the ${\Delta}so$ definitely exists in the ${\Gamma}_5$ states of the valence band of the $ZnAl_2Se_4/GaAs$ epilayer. The three photocurrent peaks observed at 10 K are ascribed to the $A_1$-, $B_1$-exciton for n = 1 and $C_{21}$-exciton peaks for n = 21.

Cyclic fatigue resistance, torsional resistance, and metallurgical characteristics of M3 Rotary and M3 Pro Gold NiTi files

  • Pedulla, Eugenio;Lo Savio, Fabio;La Rosa, Giusy Rita Maria;Miccoli, Gabriele;Bruno, Elena;Rapisarda, Silvia;Chang, Seok Woo;Rapisarda, Ernesto;La Rosa, Guido;Gambarini, Gianluca;Testarelli, Luca
    • Restorative Dentistry and Endodontics
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    • v.43 no.2
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    • pp.25.1-25.10
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    • 2018
  • Objectives: To evaluate the mechanical properties and metallurgical characteristics of the M3 Rotary and M3 Pro Gold files (United Dental). Materials and Methods: One hundred and sixty new M3 Rotary and M3 Pro Gold files (sizes 20/0.04 and 25/0.04) were used. Torque and angle of rotation at failure (n = 20) were measured according to ISO 3630-1. Cyclic fatigue resistance was tested by measuring the number of cycles to failure in an artificial stainless steel canal ($60^{\circ}$ angle of curvature and a 5-mm radius). The metallurgical characteristics were investigated by differential scanning calorimetry. Data were analyzed using analysis of variance and the Student-Newman-Keuls test. Results: Comparing the same size of the 2 different instruments, cyclic fatigue resistance was significantly higher in the M3 Pro Gold files than in the M3 Rotary files (p < 0.001). No significant difference was observed between the files in the maximum torque load, while a significantly higher angular rotation to fracture was observed for M3 Pro Gold (p < 0.05). In the DSC analysis, the M3 Pro Gold files showed one prominent peak on the heating curve and 2 prominent peaks on the cooling curve. In contrast, the M3 Rotary files showed 1 small peak on the heating curve and 1 small peak on the cooling curve. Conclusions: The M3 Pro Gold files showed greater flexibility and angular rotation than the M3 Rotary files, without decrement of their torque resistance. The superior flexibility of M3 Pro Gold files can be attributed to their martensite phase.

$CO_2$ Buffering and Hydrogen Ion Concentration Gradient across Cell Membrane in Acute Acid-Base Disturbances in Dogs (혈액과 조직의 $CO_2$완충능 및 세포막을 통한 $H^+$농도 경사)

  • Hwang, Sang-Ik;Park, Young-Bae;Min, Byoung-Ku;Kim, Woo-Gyeum
    • The Korean Journal of Physiology
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    • v.17 no.2
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    • pp.119-124
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    • 1983
  • The in vivo and in vitro buffer capacities of true plasma and tissue buffer capaciies were compared on dogs. Intracellular pH was determined on skeletal muscle by a modification of the method of Schloerb and Grantham using $C^{14}$ DMO. The in vivo curve for plasma or extracellular fluid has a much lower slope than the in vitro curve. The in vivo slope of skeletal muscle in the dog is approximately 20 sl. The slope for skeletal muscle in vivo falls between the in vitro and in vivo slopes of true plasma. It appears that intracellular hydrogen ion varies linearly with extracellular hydrogen ion when $CO_2$ tension is changed. Both hydrogen ion gradient and Hi/He ratio vary in skeletal muscle, with an increase in $CO_2$ tension. Infusion of 0.3N HCl gave two distinct patterns, the $H_i-H_e$ gradient decreased; and it would appear that very little hydrogen ion as such penetrated to the inside of the cells during the time of observation. Although lactic acid presumably enters the cell and the same of larger load was given as was used for hydrochloric acid, only very mild intracellular acidosis resulted, ostensibly due to metabolism of this substrate. Gluconic acid produced a more severe acidosis, both intracellularly and extracellularly, but with both of these acids the hydrogen ion gradient decreased and the $H_i/H_e$ ratio also decreased. The experiments on the dogs with hemorrhagic shock the hydrogen ion increase producing the acidosis originates inside the cells. Even so, the hydrogen ion gradient increased only very slightly in the acute experiments. This may suggest that even over short intervals of time skeletal muscle cells have a capacity to pump out hydrogen ions at a rate which maintains approximately the normal $H_i/H_e$ gradient when the source of the hydrogen ion is in the interior of the cell.

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A Study About Electrical Properties and Fabrication Schottky Barrirer Diode Prepared on Polar/Non-Polar of 6H-SiC (극성/무극성 6H-SiC 쇼트키 베리어 다이오드 제조 및 전기적 특성 연구)

  • Kim, Kyung-Min;Park, Sung-Hyun;Lee, Won-Jae;Shin, Byoung-Chul
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.23 no.8
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    • pp.587-592
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    • 2010
  • We have fabricated schottky barrier diode (SBDs) using polar (c-plane) and non polar (a-, m-plane) n-type 6H-SiC wafers. Ni/SiC ohmic contact was accomplished on the backside of the SiC wafers by thermal evaporation and annealed for 20minutes at $950^{\circ}C$ in mixture gas ($N_2$ 90% + $H_2$ balanced). The specific contact resistance was $3.6{\times}10^{-4}{\Omega}cm^2$ after annealing at $950^{\circ}C$. The XRD results of the alloyed contact layer show that formation of $NiSi_2$ layer might be responsible for the ohmic contact. The active rectifying electrode was formed by the same thermal evaporation of Ni thin film on topside of the SiC wafers and annealed for 5 minutes at $500^{\circ}C$ in mixture gas ($N_2$ 90% + $H_2$ balanced). The electrical properties of SBDs have been characterized by means of I-V and C-V curves. The forward voltage drop is about 0.95 V, 0.8 V and 0.8 V for c-, a- and m-plane SiC SBDs respectively. The ideality factor (${\eta}$) of all SBDs have been calculated from log(I)-V plot. The values of ideality factor were 1.46, 1.46 and 1.61 for c-, a- and m-plane SiC SBDs, respectively. The schottky barrier height (SBH) of all SBDs have been calculated from C-V curve. The values of SBH were 1.37 eV, 1.09 eV and 1.02 eV for c-, a- and m-plane SiC SBDs, respectively.