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Development and Validation of the Determination of Sorafenib in Human Plasma using Tandem Mass Spectrometry Coupled with Liquid Chromatography (고속액체크로마토그래피 텐덤질량분석기법을 이용한 사람 혈장 내 소라페닙 농도분석법의 개발 및 검정)

  • Park, Daejin;Lee, Sunggon;Kim, Woomi
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1456-1462
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    • 2012
  • Sorafenib is a multikinase inhibitor and an oral anticancer drug approved for the treatment of patients with advanced renal cell carcinoma and those with unresectable hepatocellular carcinoma. The purpose of this study was to develop an efficient method of the determination of sorafenib in human plasma using tandem mass spectrometry coupled with liquid chromatography (LC/MS/MS) and validate the method by the guidelines of the Korean Food and Drug Administration (KFDA). Plasma samples ($100{\mu}l$) were added with chlorantraniliprole as an internal standard and then mixed with the 0.1% formic acid-containing extraction solution composed of isopropyl alcohol and ethyl acetate (1:4, v/v). After centrifugation, the supernatant was concentrated at $45^{\circ}C$ under negative pressure and centrifugal force. The residue was reconstituted with a mobile phase and injected into the HPLC instrument using a reverse phase Waters XTerra$^{TM}$ C18 column (particle size $3.5{\mu}m$). Liquid chromatography was carried out within the run time of 5 min using a mobile phase composed of buffer (0.1% formic acid and 10 mM ammonium formate), methanol, and acetonitrile (1:6:3, v/v/v). The analytes were monitored by tandem mass spectrometry in the multiple reaction monitoring method programmed to detect sorafenib at 'm/z 465.2 ${\rightarrow}$ 252.5' and chlorantraniliprole at 'm/z 484.4 ${\rightarrow}$ 286.2' with positive electrospray ionization mode ($ES^+$). The result showed the proper linearity ($r^2$ > 0.99) over the range of 2,000-5,000 ng/ml with good accuracy (90.7-103.9%) and precision (less than 10%). The newly developed method using LC/MS/MS was validated by the guideline of KFDA and identified as more sensitive compared to the previous methods.

Effects of Fertilization Time and Culture Medium of Pig Oocytes Matured In Vitro by liquid Boar Sperm Stored at $4^{\circ}C$ (체외성숙된 돼지난포란을 $4^{\circ}C$ 보존 액상정액으로 체외수정시 수정시간과 배양배지의 영향)

  • Park, C. S.;Y. J. Yi;Kim, M. Y.;Y. J. Chang;Lee, S. H.;D. I. Jin
    • Korean Journal of Animal Reproduction
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    • v.27 no.3
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    • pp.215-223
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    • 2003
  • This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

Structural Properties of Social Network and Diffusion of Product WOM: A Sociocultural Approach (사회적 네트워크 구조특성과 제품구전의 확산: 사회문화적 접근)

  • Yoon, Sung-Joon;Han, Hee-Eun
    • Journal of Distribution Research
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    • v.16 no.1
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    • pp.141-177
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    • 2011
  • I. Research Objectives: Most of the previous studies on diffusion have concentrated on efficacy of WOM communication with the use of variables at individual level (Iacobucci 1996; Midgley et al. 1992). However, there is a paucity of studies which investigated network's structural properties as antecedents of WOM from the perspective of consumers' sociocultural propensities. Against this research backbone, this study attempted to link the network's structural properties and consumer' WOM behavior on cross-national basis. The major research objective of this study was to examine the relationship between network properties and WOM by comparing Korean and Chinese consumers. Specific objectives of this research are threefold; firstly, it sought to examine whether network properties (i.e., tie strength, centrality, range) affect WOM (WOM intention and quality of WOM). Secondly, it aimed to explore the moderating effects of cutural orientation (uncertainty avoidance and individuality) on the relationship between network properties and WOM. Thirdly, it substantiates the role of innovativeness as antecedents to both network properties and WOM. II. Research Hypotheses: Based on the above research objectives, the study put forth the following research hypotheses to validate. ${\cdot}$ H 1-1 : The Strength of tie between two counterparts within network will positively influence WOM effectivenes ${\cdot}$ H 1-2 : The network centrality will positively influence the WOM effectiveness ${\cdot}$ H 1-3 : The network range will positively influence the WOM effectiveness ${\cdot}$ H 2-1 : The consumer's uncertainty avoidance tendency will moderate the relationship between network properties and WOM effectiveness ${\cdot}$ H 2-2 : The consumer's individualism tendency will moderate the relationship between network properties and WOM effectiveness ${\cdot}$ H 3-1 : The consumer's innovativeness will positively influence the social network properties ${\cdot}$ H 3-2 : The consumer's innovativeness will positively influence WOM effectiveness III. Methodology: Through a pilot study and back-translation, two versions of questionnaire were prepared, one in Korean and the other in Chinese. The chinese data were collected from the chinese students enrolled in language schools in Suwon city in Korea, while Korean data were collected from students taking classes in a major university in Seoul. A total of 277 questionnaire were used for analysis of Korean data and 212 for Chinese data. The reason why Chinese students living in Korea rather than in China were selected was based on two factors: one was to neutralize the differences (ie, retail channel availability) that may arise from living in separate countries and the second was to minimize the difference in communication venues such as internet accessibility and cell phone usability. SPSS 12.0 and AMOS 7.0 were used for analysis. IV. Results: Prior to hypothesis verification, mean differences between the two countries in terms of major constructs were performed with the following result; As for network properties (tie strength, centrality and range), Koreans showed higher scores in all three constructs. For cultural orientation traits, Koreans scored higher only on uncertainty avoidance trait than Chinese. As a result of verifying the first research objective, confirming the relationship between network properties and WOM effectiveness, on Korean side, tie strength(Beta=.116; t=1.785) and centrality (Beta=.499; t=6.776) significantly influenced on WOM intention, and similar finding was obtained for Chinese side, with tie strength (Beta=.246; t=3.544) and centrality (Beta=.247; t=3.538) being significant. However, with regard to WOM argument quality, Korean data yielded only centrality (Beta=.82; t=7.600) having a significant impact on WOM, whereas China showed both tie strength(Beat=.142; t=2.052) and centrality(Beta=.348; t=5.031) being influential. To answer for the second research objective addressing the moderating role of cultural orientation, moderated regression anaylsis was performed and the result showed that uncertainty avoidance moderated between network range and WOM intention for both Korea and China, But for Korea, the uncertainty avoidance moderated between tie strength and WOM quality, while for China it moderated between network range and WOM intention. And innovativeness moderated between tie strength and WOM intention for Korea but it moderated between network range and WOM intention for China. As a result of analysing for third research objective, we found that for Korea, innovativeness positively influenced centrality only (Beta=.546; t=10.808), while for China it influenced both tie strength (Beta=.203; t=2.998) and centrality(Beta=.518; t=8.782). But for both countries alike, the innovativeness influenced positively on WOM (WOM intention and WOM quality). V. Implications: The study yields the two practical implications. Firstly, the result suggests that companies targeting multinational customers need to identify segments which are susceptible to the positive WOM and WOM information based on individual traits such as uncertainty avoidance and individualism and based on that, develop marketing communication strategy. Secondly, the companies need to divide the market on Roger's five innovation stages and based on this information, enforce marketing strategy which utilizes social networking tools such as public media and WOM. For instance, innovator and early adopters, if provided with new product information, will be able to capitalize upon the network advantages and thus add informational value to network operations using SNS or corporate blog.

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