• Title/Summary/Keyword: freeze-thaw stability

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Evaluation of the stability of IgM and specific antibody response of sevenband grouper Epinephelus septemfasciatus for application of antibody-detection ELISA (항체검출 ELISA 적용을 위한 능성어 IgM의 안정성 및 특이 항체 반응 평가)

  • Kim, Chun-Seob;Jang, Min-Seok;Kim, Wi-Sik;Kim, Jong-Oh;Kim, Du-Woon;Kim, Do-Hyung;Han, Hyun-Ja;Jeong, Sung-Ju;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.335-342
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    • 2009
  • The stability of immunoglobulin M (IgM) on different serum storage conditions and specific antibody response were tested using the serum collected from sevenband grouper Epinephelus septemfasciatus by enzyme-linked immunosorbent assay (ELISA). To test the effect of storage temperature and duration, sevenband grouper antiserum against bovine serum albumin (BSA) was stored at -80, -20 or 4${^{\circ}C}$ for 1, 34, 61 or 119 days. In addition, to test the effect of repeated freeze-thawing condition, the anti-BSA fish serum was frozen at -20 and -80${^{\circ}C}$ and then thawn and frozen for 1, 5 or 10 times repeatedly. Consequently, no significant difference was found in ELISA optical density (O.D.) values of sera for the above mentioned storage conditions: different temperatures (-80, -20 and 4${^{\circ}C}$), durations of storage (1, 34, 61 and 119 days), and repeated thaw-freeze cycles (1, 5, and 10 times), indicating that IgMs of test fish were stable. The specific antibody response of sevenband grouper was observed after BSA-immunization of the test fish reared at 20 ${^{\circ}C}$ or 25${^{\circ}C}$. At the rearing temperature of 20${^{\circ}C}$, the specific antibody against BSA first appeared at 14 days and maximum antibody titer was observed between 21 and 28 days, while at the rearing temperature of 25 ${^{\circ}C}$, specific antibody appeared at 7 days and maximum antibody titer was observed between 14 and 21 days. In conclusion, the rearing temperature at 25${^{\circ}C}$ gave a faster and higher specific antibody response than at 20${^{\circ}C}$ and the specific antibody response maintained for approximately 2 months at 20℃ and 25${^{\circ}C}$.

Development and Validation of an HPLC Method for the Pharmacokinetic Study of Fexofenadine in Human (테르페나딘 체내동태 연구를 위한 혈청 중 펙소페나딘의 HPLC 정량법 개발 및 검증)

  • Cho, Hye-Young;Kang, Hyun-Ah;Kim, Yoon-Gyoon;Choi, Hoo-Kyun;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.35 no.6
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    • pp.437-443
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    • 2005
  • A rapid, selective and sensitive reversed-phase HPLC method for the determination of a major metabolite of terfenadine, fexofenadine, in human serum was developed, validated, and applied to the pharmacokinetic study of terfenadine. Fexofenadine and internal standard, haloperidol were extracted from human serum by liquid-liquid extraction with acetonitrile and analyzed on a $Symmetry^{TM}$ C8 column with the mobile phase of 1% triethylamine phosphate (pH 3.7)-acetonitrile (67:33, v/v, adjusted to pH 5.6 with triethylamine). Detection wavelength of 230 nm for excitation, 280 nm for emission and flow rate of 1.0 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed fexofenadine concentration (50 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 10-500 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of serum was 10 ng/mL, which was sensitive enough for the pharmacokinetic studies of terfenadine. The overall accuracy of the quality control samples ranged from 95.70 to 114.58% for fexofenadine with overall precision (% C.V.) being 3.53-14.39%. The relative mean recovery of fexofenadine for human serum was 90.17%. Stability studies (freeze-thaw, short-term, extracted serum sample and stock solution) showed that fexofenadine was stable during storage, or during the assay procedure in human serum. However, the storage at $-70^{\circ}C$ for 4 weeks showed that fexofenadine was not stable. The peak area and retention time of fexofenadine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of fexofenadine in human serum samples for the pharmacokinetic studies of orally administered Tafedine tablet (60 mg as terfenadine) at three different laboratories, demonstrating the suitability of the method.

Development and Validation of an HPLC Method for the Pharmacokinetic Study of Fenoprofen in Human (페노프로펜 체내동태 연구를 위한 혈청 중 페노프로펜의 HPLC 정량법 개발 및 검증)

  • Cho, Hye-Young;Kang, Hyun-Ah;Kim, Yoon-Gyoon;Sah, Hong-Kee;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.35 no.6
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    • pp.423-429
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    • 2005
  • A selective and sensitive reversed-phase HPLC method for the determination of fenoprofen in human serum was developed, validated, and applied to the pharmacokinetic study of fenoprofen calcium. Fenoprofen and internal standard, ketoprofen, were extracted from human serum by liquid-liquid extraction with diethyl ether and analyzed on a Luna C18(2) column with the mobile phase of acetonitrile-3 mM potassium dihydrogen phosphate (32:68, v/v, adjusted to pH 6.6 with phosphoric acid). Detection wavelength of 272 nm and flow rate of 0.25 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed fenoprofen concentration $(2\;{\mu}g/mL)$ with respect to its peak area and retention time. And also, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of $0.05-100\;{\mu}g/mL$ with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was $0.05\;{\mu}g/mL$, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 92.27 to 109.20% for fenoprofen with overall precision (% C.V.) being 5.51-11.71 %. The relative mean recovery of fenoprofen for human serum was 81.7%. Stability (freeze-thaw, short and long-term) studies showed that fenoprofen was not stable during storage. But, extracted serum sample and stock solution were allowed to stand at ambient temperature for 12 hr prior to injection without affecting the quantification. The peak area and retention time of fenoprofen were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of fenoprofen in human serum samples for the pharmacokinetic studies of orally administered Fenopron tablet (600 mg as fenoprofen) at three different laboratories, demonstrating the suitability of the method.

Development and Validation of an HPLC Method for the Pharmacokinetic Study of Pentoxifylline in Human Serum (체내동태 연구를 위한 혈청 중 펜톡시필린의 HPLC 정량법 개발 및 검증)

  • Cho, Hea-Young;Kang, Hyun-Ah;Yoo, Hee-Doo;Lee, Hwa-Jeong;Moon, Jai-Dong;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.2
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    • pp.89-95
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    • 2006
  • A selective and sensitive reversed-phase HPLC method for the determination of pentoxifylline in human serum was developed, validated, and applied to the pharmacokinetic study of pentoxifylline. Pentoxifylline and internal standard, chloramphenicol, were extracted from the serum by liquid-liquid extraction with dichloromethane and analyzed on a Luna CI8(2) column with the mobile phase of acetonitrile-0.034 M phosphoric acid (25:75, v/v, adjusted to pH 4.0 with 10 M NaOH). Detection wavelength of 273 nm and flow rate of 0.8 mL/min were used. This method showed linear response over the concentration range of 10-500 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of the serum was 10 ng/mL, which was sensitive enough for pharmacokinetic studies of pentoxifylline. The overall accuracy of the quality control samples ranged from 89.3 to 92.7% for pentoxifylline with overall precision (% C.V.) being 4.1-9.2%. The relative mean recovery of pentoxifylline for human serum was 105.8%. Stability (stock solution, short and long-term) studies showed that pentoxifylline was not stable during storage. But three freeze-thaw cycles and extracted serum samples were stable. This method showed good ruggedness (within 15% C.V.) and was successfully applied for the analysis of pentoxifylline in human serum samples for the pharmacokinetic studies of orally administered $Trental^{\circledR}$ tablet (400 mg pentoxifylline), demonstrating the suitability of the method.

Numerical Study on Freezing and Thawing Process in Modular Road System (모듈러 도로시스템의 동결-융해에 대한 수치해석적 연구)

  • Shin, Hosung;Kim, Jinwook;Lee, Jangguen;Kim, Dong-Gyou
    • Journal of the Korean Geotechnical Society
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    • v.33 no.3
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    • pp.49-62
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    • 2017
  • In order to understand response of geo-structures to the freezing-thawing process in the ground, it is necessary to consider phase change of the pore water of the ground and also to understand soil interaction with structures. In this study, numerical analysis was carried out for freezing and thawing effect on the modular road system. Neumann's theoretical equation for freezing-thawing processes in porous media can be used to estimate frozen depth and heaving from basic soil properties and ground and surface temperature, but its application is limited to the case for the sediment with fully saturated condition and zero unfrozen water content. Numerical analysis of the modular road system was performed on various soil types and different ground water table as the varying freezing index. The amount of heaving in the silty soil was much larger than those in granite weathered soil or sandy soil, and lowering groundwater level reduced ground heaving induced by freezing. Numerical analysis for temperature history of the ground surface predicted residual heaving near the surface by the freeze-thaw process in silty soil. It ought to reduce stiffness and bearing capacity of the ground so that it will impair stability and serviceability of new road system. However, the amount of residual heaving was insignificant for the road system installed in weathered soil granite and sandy soil. Since modular road system is a pavement structure mounted on the supporting substructure unlike the prevalent road pavement system, strict criteria should be applied for uniform and differential settlement of the pavement system.

Evaluation of Field Applicability with Coal Mine Drainage Sludge (CMDS) as a Liner: Part I: Physico-Chemical Characteristics of CMDS and a Mixed Liner (차수재로의 광산슬러지 재활용 적용성 평가: Part I: 광산배수슬러지 및 혼합차수재의 물리·화학적 성질)

  • Lee, Jai-Young;Bae, Sun-Young;Woo, Seung-Hyun
    • Journal of the Korean Geosynthetics Society
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    • v.10 no.2
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    • pp.67-72
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    • 2011
  • CMDS (Coal Mine Drainage Sludge) is mainly generated from acid mine drainage during physicochemical treatment or electrical purification. CMDS is well worth considering on recycling possibilities in various areas. This research applies the liner and cover materials using waste disposal landfill generally to treat acid mine drainage sludge. In this Part I of the two parts paper, physico-chemical characteristics of CMDS, bentonite and cement to prepare the liner have been identified using XRD, XRF, FESEM. In addition, combining their physicochemical characteristics, the optimum mixing ratio has been determined to be 1: 0.5: 0.3 for CMDS: bentonite: cement by the batch tests. Initial permeability of CMDS was $7.10{\times}10^{-7}cm/s$. Through the leaching test, it was confirmed that its mixture was environmentally safe. In the Part 2, a large-scale Lysimeter was used to simulate the effects of the layer on the freeze/thaw for evaluation on field applicability and stability.

Development and Validation of HPLC Method for Pharmacokinetic Study of Promethazine in Human (염산프로메타진 체내동태 연구를 위한 혈청 중 프로메타진의 HPLC 정량법 개발 및 검증)

  • Cho, Hae-Young;Kang, Hyun-Ah;Lee, Hwa-Jeong;Choi, Hoo-Kyun;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.1
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    • pp.23-29
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    • 2006
  • A rapid, selective and sensitive reversed-phase HPLC method for the determination of promethazine in human serum was developed, validated, and applied to the pharmacokinetic study of promethazine. Promethazine and internal standard, chlorpromazine, were extracted from human serum by liquid-liquid extraction with n-hexane containing 0.8% isopropanol and analyzed on a Capcell Pak CN column with the mobile phase of acetonitrile-0.2 M potassium dihydrogen phosphate (42:58, v/v, adjusted to pH 6.0 with 1 M NaOH). Detection wavelength of 251 nm and flow rate of 0.9 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed promethazine concentration (10 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 1-40 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was 1 ng/mL, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 96.15 to 105.40% for promethazine with overall precision (% C.V.) being 6.70-11.22%. The relative mean recovery of promethazine for human serum was 63.54%. Stability (freeze-thaw and short-term) studies showed that promethazine was stable during storage, or during the assay procedure in human serum. However, the storage at $-80^{\circ}C$ for 4 weeks showed that promethazine was not stable. Extracted serum sample and stock solution were not allowed to stand at ambient temperature for 12 hr prior to injection. The peak area and retention time of promethazine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of promethazine in human serum samples for the pharmacokinetic studies of orally administered Himazin tablet (25 mg as promethazine hydrochloride) at three different laboratories, demonstrating the suitability of the method.

Validated HPLC Method for the Pharmacokinetic Study of Atenolol and Chlorthalidone Combination Therapy in Korean Subjects

  • Kang, Hyun-Ah;Kim, Hwan-Ho;Kim, Se-Mi;Yoon, Hwa;Cho, Hea-Young;Oh, Seaung-Youl;Choi, Hoo-Kyun;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.5
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    • pp.331-338
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    • 2006
  • A rapid, selective and sensitive reverse-phase HPLC methods for the determination of atenolol and chlorthalidone in human serum and whole blood were validated, and applied to the pharmacokinetic study of atenolol and chlorthalidone combination therapy. Atenolol and an internal standard, pindolol, were extracted from human serum by liquid-liquid extraction, and analyzed on a $\mu$-Bondapak C18 $10-{\mu}$ column in a mobile phase of methanol-0.01 M potassium dihydrogenphosphate(30:70, v/v, adjusted to pH 3.5) and fluorescence detection(emission: 300 nm, excitation: 224 nm). Chlorthalidone and an internal standard, probenecid, were extracted form human whole blood by liquid-liquid extraction, and analyzed on a Luna C18 $5-{\mu}$ column in a mobile phase of acetonitrile containing 77% 0.01 M sodium acetate and UV detection at 214 nm. These analysis were performed at three different laboratories using the same quality control(QC) samples. The chromatograms showed good resolution, sensitivity, and no interference by human serum and whole blood, respectively. The methods showed linear responses over a concentration range of 10-1,000 ng/mL for atenolol and 0.05-20 ${\mu}g/mL$ for chlorthalidone, with correlation coefficients of greater than 0.999 at all the three laboratories. Intra- and inter-day assay precision and accuracy fulfilled international requirements. Stability studies(freeze-thaw, short-, long-term, extracted sample and stock solution) showed that atenolol and chlorthalidone were stable. The lower limit of quantitation of atenolol and chlorthalidone were 10 ng/mL and 0.05 ${\mu}g/mL$, respectively, which was sensitive enough for pharmacokinetic studies. These methods were applied to the pharmacokinetic study of atenolol and chlorthalidone in human volunteers following a single oral administration of Hyundai $Tenoretic^{\circledR}$ tablet(atenolol 50 mg and chlorthalidone 12.5 mg) at three different laboratories.

Physicochemical Properties of Commercial Sweet Potato Starches (시판 고구마전분의 이화학적 특성)

  • Baek, Man-Hee;Cha, Dong-Su;Park, Hyun-Jin;Lim, Seung-Taik
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.755-762
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    • 2000
  • Physicochemical properties of commercial sweet potato starches manufactured by 7 different companies were investigated in comparison with corn and potato starches. Crude ash and protein content varied from 0.36 to 1.02%, and from 0.04 to 0.14% based on dry weight, respectively. The protein contents were relatively smaller than that of corn or potato starch. But whiteness of the sweet potato starches was less than that of corn or potato starch. Mean diameter of the sweet potato starch granules varied from 14.23 to $21.08\;{\mu}m$ depending on the company and all sweet potato starches showed bimodal size distributions. Pasting viscosity measured by Rapid Viscoanalyzer(RVA) also showed variations among the starches of different companies. The starch from D company in Korea had the lowest pasting temperature$(74.00^{\circ}C)$ whereas the starch from a phillippine company(P) did the highest one$(80.35^{\circ}C)$. The peak viscosity of sweet potato starches was higher than that of corn starch but lower than that of potato starch. The D company starch also showed the highest peak viscosity(2283 cp) among the starches tested. Paste breakdown by hot shearing ranged from 524 cp (S company) to 1279 cp (HL company). Textural properties of the starch gels appeared significantly different among the starches of different manufacturers. The greatest hardness of the gel was $137.90\;g_{f}$ at 1 day storage whereas the lowest value was $31.53\;g_{f}$. Except the starches from 2 companies (P and S), the sweet potato starches formed very soft and weak gels. P or S company starches formed the gels similar to potato starch. Syneresis by freeze-thawing treatments appeared less for sweet potato starch gels than that for corn starch gels, but greater than that for potato starch gel. The overall properties of the sweet potato starches varied by the manufacturing companies, and ranged between those of corn and potato starches.

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