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Comparative Evaluation of the Analytical Methods used to Determine Pesticide Residues in Milk via Dispersive Solid Phase Extraction (Dispersive Solid Phase Extraction을 사용한 우유 내 잔류농약 다성분 동시 분석법 비교연구)

  • Oh, Nam Su;Shin, Yong Kook;Lee, Ji Young;Baick, Seung Chun
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.1
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    • pp.27-34
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    • 2015
  • The aim of this study was to optimize a simple, fast, and economic analytical method for the simultaneous determination of various pesticides (aldrin, p,p'-DDT, o,p'-DDT, p,p'-DDE, ${\alpha}$-endosulfan, ${\beta}$-endosulfan, dieldrin, heptachlor, permethrin, chlordane, deltamethrin, diazinon, bifenthrin, methoprene, propargite, fenpropathrin, cypermethrin, fenvalerate, and fenpropathrin) in milk by using dispersive solid phase extraction (SPE). In this study, two different extraction methods (low temperature cleanup and liquid-liquid partitioning), which were followed by a cleanup process based on dispersive-SPE, were evaluated and compared for the 19 pesticides. The results for all the pesticides were determined by gas chromatography mass spectrometry (GC/MS) with selected-ion monitoring mode, and the matrix effect of the method was evaluated. Comparison of these approaches yielded higher and more consistent recoveries of most pesticides at fortification levels of $1{\mu}g/mL$ using low-temperature fat precipitation, followed by cleanup process based on dispersive-SPE with PSA and C18 as sorbents, than other preparation process. The relative standard deviation was <20 % and the combination of this method were very effective for the cleanup.

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Production of Mozzarella Cheese Analogue by Ultrafiltration (한외여과를 이용한 Mozzarella Cheese Analogue 제조에 관한 연구)

  • Lee, Sung-Hui;Song, Kwang-Young;Seo, Kun-Ho;Yoon, Yoh-Chang
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.1
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    • pp.21-33
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    • 2013
  • The purpose of this study was to improve the production of Mozzarella cheese analogues manufactured using mixtures of soy milk and concentrated raw milk by performing ultrafiltration (UF) and to assess the quality of these cheeses during a 30-day storage period at $4^{\circ}C$, relative to that of Mozzarella cheese manufactured with the traditional method. The solid consistency of Mozzarella cheese analogue prepared from milk mixtures was lower than that of cheese manufactured from raw milk or soy milk and increased during storage, which is considered to be the result of decreasing water levels, as well as with increasing soy milk concentrations. In the Mozzarella cheese analogue generated using the milk mixtures, the fat content decreased with increase in the soy milk concentration, while it decreased during the storage period. Lactose levels were lowest in cheese composed of soy milk or raw milk and processed by UF, and decreased during storage in cheese produced using milk mixtures. In milk mixtures containing soy milk, the protein concentration increased with increasing amounts of raw milk and did not change during the storage period. The water-soluble nitrogen compound level was similar between cheeses and increased only slightly during storage. The amount of non-protein nitrogen compounds was higher in the cheese analogue than in the control cheese and tended to increase during storage. Analysis of the physicochemical traits of the Mozzarella cheese analogue yielded the following results: During storage, titratable acidity levels increased while pH tended to decrease. After analysis using electropherograms, it was classified as ${\alpha}$-, ${\beta}$-, or ${\kappa}$-casein. The results of rheometry tests showed that in the Mozzarella cheese analogue prepared from milk mixtures, with raw milk concentrated by UF, increases in concentration rate lead to lowered hardness, elasticity, cohesiveness, and brittleness. When cheese was produced from milk mixtures and concentrated by UF, meltability increased as the concentration rate increased, although to an extent that was less than that observed for the control cheese, and tended to increase during storage. Sensory evaluation showed that the analogue cheese was much better than the control cheese in terms of formation, appearance, and flavor.

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Comparison of Human Sensibility in Driving Simulator and Roller-Coaster Simulator (자동차 시뮬레이터와 롤러코스터 시뮬레이터 주행에 따른 감성 비교)

  • 민병찬;전효정;강인형;성은정;김철중;윤석준
    • Science of Emotion and Sensibility
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    • v.6 no.3
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    • pp.13-20
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    • 2003
  • In an experimental study, we assessed human sensibility in terms of psychophysiological response according to change of speeds (40, 70, 100km/h) in a driving simulator and application of motion fitters (washout filter, non-washout later) in a roller coaster simulator. For the driving and roller coaster simulators, a group of 12 healthy men in their twenties and a group of 8 healthy men in their twenties, respectively, participated. Participants each completed a simulator sickness questionnaire (SSQ), a subjective assessment of sensations of pleasantness, tension, and arousal, and perception of speed. Physiological signals were measured by 1/f fluctuation of EEG (electroencephalogram), ECG (electrocardiogram), and GSR (galvanic skin response). These were measured pre-to-post under the experimental conditions for each simulator. Subjective pleasantness, tension, arousal, and perception of speed and physiological responses indicating a feeling of pleasantness by 1/f fluctuation were higher for the roller coaster simulator than those measured for low speed driving in the driving simulator. The mean frequency of alpha band (8-l3㎐) in EEG increased with exposure to the driving simulator relative to that for the roller coaster simulator. Heart rate variability and GSR were significantly changed between pre- and post- under each condition in the driving and roller coaster simulators. The data suggest that subjective sensibility was elevated according to gain of speed and variety of simulator motion, and physiological responses were activated with increased speed.

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The Prediction of the Expected Current Selection Coefficient of Single Nucleotide Polymorphism Associated with Holstein Milk Yield, Fat and Protein Contents

  • Lee, Young-Sup;Shin, Donghyun;Lee, Wonseok;Taye, Mengistie;Cho, Kwanghyun;Park, Kyoung-Do;Kim, Heebal
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.1
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    • pp.36-42
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    • 2016
  • Milk-related traits (milk yield, fat and protein) have been crucial to selection of Holstein. It is essential to find the current selection trends of Holstein. Despite this, uncovering the current trends of selection have been ignored in previous studies. We suggest a new formula to detect the current selection trends based on single nucleotide polymorphisms (SNP). This suggestion is based on the best linear unbiased prediction (BLUP) and the Fisher's fundamental theorem of natural selection both of which are trait-dependent. Fisher's theorem links the additive genetic variance to the selection coefficient. For Holstein milk production traits, we estimated the additive genetic variance using SNP effect from BLUP and selection coefficients based on genetic variance to search highly selective SNPs. Through these processes, we identified significantly selective SNPs. The number of genes containing highly selective SNPs with p-value <0.01 (nearly top 1% SNPs) in all traits and p-value <0.001 (nearly top 0.1%) in any traits was 14. They are phosphodiesterase 4B (PDE4B), serine/threonine kinase 40 (STK40), collagen, type XI, alpha 1 (COL11A1), ephrin-A1 (EFNA1), netrin 4 (NTN4), neuron specific gene family member 1 (NSG1), estrogen receptor 1 (ESR1), neurexin 3 (NRXN3), spectrin, beta, non-erythrocytic 1 (SPTBN1), ADP-ribosylation factor interacting protein 1 (ARFIP1), mutL homolog 1 (MLH1), transmembrane channel-like 7 (TMC7), carboxypeptidase X, member 2 (CPXM2) and ADAM metallopeptidase domain 12 (ADAM12). These genes may be important for future artificial selection trends. Also, we found that the SNP effect predicted from BLUP was the key factor to determine the expected current selection coefficient of SNP. Under Hardy-Weinberg equilibrium of SNP markers in current generation, the selection coefficient is equivalent to $2^*SNP$ effect.

Anti-inflammatory Effects of Illicium verum Hook. f. via Suppression of NFκB Pathway (NF-κB 억제를 통한 대회향(大茴香)의 항염증효능)

  • Keum, Soo Yeon;Park, Sang Mi;Jegal, Kyung Hwan;Hwangbo, Min;Cho, Il Je;Park, Chung A;Kim, Sang Chan;Jee, Seon Young
    • Herbal Formula Science
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    • v.24 no.4
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    • pp.243-257
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    • 2016
  • Objectives : Illicium verum Hook. f. has been known to possess antimicrobial, antioxidant, antifungal, anti-inflammatory, insecticidal, analgesic, sedative, convulsive activities, it has been rarely conducted to evaluate the immuno-biological activity. The present study was examined to evaluate the anti-inflammatory effects of the Illicium verum Hook. f. water extracts (IVE) in vivo and in vitro. Methods : Cell viability was measured by MTT assay. The relative levels of NO were measured with Griess reagent. iNOS, COX-2, $NF-{\kappa}B$ and target proteins were detected by immunoblot analysis, and levels of cytokines were analyzed by ELISA kit. Anti-edema effect was determined in the carrageenan (CA)-induced paw edema model in rats. Results : All dosages of IVE used in MTT assay had no significant cytotoxicity. The increases of NO production and iNOS expression were detected in LPS-treated cells compared with control. However, these increases were attenuated by treatment with IVE. Also, IVE reduced the elevated production of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 by LPS. IVE inhibited the $p-I{\kappa}B$ and translocation of $NF-{\kappa}B$ to nuclear. Furthermore, IVE significantly inhibited the increases of hind paw swelling, skin thicknesses and inflammatory cell infiltrations induced by CA injection. Therefore, IVE will be favorably inhibited the acute edematous inflammations. Conclusion : These results provide evidences that anti-inflammatory effect of IVE is partly due to the reduction of some inflammatory mediators by suppression of $NF-{\kappa}B$ pathway.

Estrogenic Activity, and Developmental Toxicity Studies of Pyrethroid Insecticides

  • Kim, Soon-Sun;Rhee, Gyu-Seek;Lee, Rhee-Da;Kwack, Seung-Jun;Lim, Kwon-Jo;Yhun, Hyo-Jung;Park, Kui-Lea
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.57-69
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    • 2003
  • It is well known that many pesticides possess hormonal activity, and affect the developments of wildlife and mammals including human. Currently, pyrethroid insecticides are in worldwide use to control in and outdoor pests, providing potential far environmental exposure. Hormonal activities of these pyrethroid insecticides, however, have been little studied, and the developmental effects of them were no reported. Therefore, we firstly examined the potential estrogenic activities of some pyrethroid insecticides (permethrin, cypermethrin, tetramethrin, deltamethrin, sumithrin, fenvalerate and bioallethrin) by immature rat uterotrophic assay, luciferase reporter gene assay and Calbindin-D$\sub$9k/ (CaBP-9k) gene expression assay. Uterine wet weights were increased by permethrin and the permethrin-induced weights were inhibited by ICI 182780 in the uterolrophic assay. On the other hand tetramethrin significantly reduced uterine and vaginal wet weights, and also inhibited the E2-induced weight increases at all doses tested. Cypermethrin and sumithrin had a tendency to increase uterine weights, although not statistically significant. Permethrin and cypermethrin dose-dependently increased the luciferase activity in reporter gene assay. Northern blot analysis showed that permethrin induced CaBP-9k mRNA expression whereas tetramethrin inhibted. Subsequent studies were conducted to investigate the possible developmental effects of four pyrethroid insecricides (permethrin, cypermethrin, sumithrin and teramethrin). Either diethlbestrol (DES) or 17${\beta}$ -estradiol (E2) was used as a reference control in this study. Pyrethroid insecticides were administered to Sprague Dawley rats via subcutaneous injection at 6 to 18 days of gestation or 1 to 5 days after birth. In utero treatment of permethrin (10mg/kg/day) in female rat resulted in significant increases in uterine and ovarian weights while significant decreases in serum E2 concentration, uterine and ovarian ER${\alpha}$ mRNA levels. Sumithrin and permethrin led to acceleration in vaginal opening of female rat, while delay in preputial separation of male after neonatal treatment. Anogenital distances of PND 18 were significantly reduced in sumthrin-treated, and permerhrin-treated male rats after neonatal treatment. All the pyrethroid insecticides tested caused significant increases in uterine weights on PND 18, while significant reductions in the first diestrus phase when neonataly treated. In addition, exposure to pyrethroids in neonatal period led to significant reduction in relative brain weight in female rat on PND 18, but its weight was recovered in diestrus phase. In summary, Our experimental data demonstrate the possibilities of developmental effects of pyrethroid insecticides via estrogenic or antiestrogenic activity.

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Variants of Inflammnation-related Genes and the Risk of Gallstones and Biliary Tract Cancer: A population-baged Study in China

  • Hsing, Ann W.;Sakoda, Lori;Chen, Jin-Bo;Rashid, Asif;Wang, Bin-Shen;Shen, Ming-Chang;Chen, Eric;Rosenberg, Phillip;Zhang, Mingdong;Andreotti, Gabriella;Welch, Robert;Yeager, Meredith;Fraumeni Jr. Joseph F.;Gao, Yu-Tang;Stephen J. Chanock
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2006.11a
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    • pp.32-33
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    • 2006
  • There is compelling evidence that chronic inflammation predisposes to biliary tract cancer. Previously we found that aspirin use and variants in the PTGS2 gene, both of which are closely linked to inflammation, were associated with biliary tract cancer risk in a population-based study in China. To test the inflammation hypothesis further, we examined the associations of variants in 20 genes involved in the inflammation pathway with risk of biliary tract cancer and stones in a large population-based case-control study in Shanghai, China. We genotyped 56 single nucleotide polymorphisms (SNPs)from 20 inflammation genes in 411 biliary tract cancer cases (237 gallbladder cancers, 127 extrahepatic bile duct cancers, and 47 ampullary cancers), 895 subjects with biliary stones, and 786 population controls. Unconditional logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (Cls) for the association of individual SNPs and haplotypes with biliary stones and biliary tract cancer risk. Of the 56 SNPs examined, 20 showed some associations with biliary cancer and stones. Specifically, variants of the IL8, IL8RB, RNASEL, TGF-beta, and TNF-alpha genes were associated with gallstone risk, while variants in the IL1A, IL10, VEGF, and RNASEL genes were associated with gallbladder cancer risk. Adjustment for multiple comparisons did not materially change these results. Of the 10 genes with multiple SNPs, we inferred halotypes; only one haplotype in the IL8RBgene was associated with gallstones. The haplotype frequency was significantly different between bile dict cancer cases and control (p=0.007). A haplotype comprising 3 SNPs in the IL8RB gene (rs2230054, rs1126579, rs1126580) was associated with a 54% increased risk of bile duct stones (95% CI 1.14-2.07, p=0.02), relative to the most frequent haplotype. In summary, common variants in immune-related genes influencing inflammatory responeses were associated with gallstones and biliary tract cancer, lending further support to the role of inflammation in the pathogenesis of biliary stones and biliary tract cancer. Future larger studies with more complete gene coverage are needed to confirm these results.

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The Role and Regulation of MCL-1 Proteins in Apoptosis Pathway

  • Bae, Jeehyeon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2002.07a
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    • pp.113-113
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    • 2002
  • Phylogenetically conserved Bcl-2 family proteins play a pivotal role in the regulation of apoptosis from virus to human. Members of the Bcl-2 family consist of antiapoptotic proteins such as Bcl-2, Bcl-xL, and Bcl-w, and proapoptotic proteins such as BAD, Bax, BOD, and Bok. It has been proposed that anti- and proapoptotic Bcl-2 proteins regulate cell death by binding to each other and forming heterodimers. A delicate balance between anti- and proapoptotic Bcl-2 family members exists in each cell and the relative concentration of these two groups of proteins determines whether the cell survives or undergoes apoptosis. Mcl-1 (Myeloid cell :leukemia-1) is a member of the Bcl-2 family proteins and was originally cloned as a differentiation-induced early gene that was activated in the human myeloblastic leukemia cell line, ML-1 . Mcl-1 is expressed in a wide variety of tissues and cells including neoplastic ones. We recently identified a short splicing variant of Mcl-1 short (Mcl-IS) and designated the known Mcl-1 as Mcl-1 long (Mcl-lL). Mcl-lL protein exhibits antiapoptotic activity and possesses the BH (Bcl-2 homology) 1, BH2, BH3, and transmembrane (TM) domains found in related Bcl-2 proteins. In contrast, Mcl-1 S is a BH3 domain-only proapoptotic protein that heterodimerizes with Mcl-lL. Although both Mc1-lL and Mcl-lS proteins contain BH domains fecund in other Bcl-2 family proteins, they are distinguished by their unusually long N-terminal sequences containing PEST (proline, glutamic acid, serine, and threonine) motifs, four pairs of arginine residues, and alanine- and glycine-rich regions. In addition, the expression pattern of Mcl-1 protein is different from that of Bcl-2 suggesting a unique role (or Mcl-1 in apoptosis regulation. Tankyrasel (TRF1-interacting, ankyrin-related ADP-related polymerasel) was originally isolated based on its binding to TRF 1 (telomeric repeat binding factor-1) and contains the sterile alpha motif (SAM) module, 24 ankyrin (ANK) repeats, and the catalytic domain of poly(adenosine diphosphate-ribose) polymerase (PARP). Previous studies showed that tankyrasel promotes telomere elongation in human cells presumably by inhibiting TRFI though its poly(ADP-ribosyl)action by tankyrasel . In addition, tankyrasel poly(ADP-ribosyl)ates Insulin-responsive amino peptidase (IRAP), a resident protein of GLUT4 vesicles, and insulin stimulates the PARP activity of tankyrase1 through its phosphorylation by mitogen-activated protein kinase (MAPK). ADP-ribosylation is a posttranslational modification that usually results in a loss of protein activity presumably by enhancing protein turnover. However, little information is available regarding the physiological function(s) of tankyrase1 other than as a PARP enzyme. In the present study, we found tankyrasel as a specific-binding protein of Mcl-1 Overexpression of tankyrasel led to the inhibition of both the apoptotic activity of Mel-lS and the survival action of Mcl-lL in mammalian cells. Unlike other known tankyrasel-interacting proteins, tankyrasel did not poly(ADP-ribosyl)ate either of the Mcl-1 proteins despite its ability to decrease Mcl-1 proteins expression following coexpression. Therefore, this study provides a novel mechanism to regulate Mcl-1-modulated apoptosis in which tankyrasel downregulates the expression of Mcl-1 proteins without the involvement of its ADP-ribosylation activity.

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A Survey on the Recognition of Korean Farmers for the Safety of Environment-friendly Organic Materials (친환경유기농자재 안전성 확보를 위한 농업인의 인식도 조사)

  • Paik, Min-Kyoung;Lee, Je-Bong;Oh, Jin-A;Kim, Min-Ju;Kim, Sang-Su;Choi, Chil-Gu;Kim, Do-Hoo
    • Korean Journal of Organic Agriculture
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    • v.21 no.2
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    • pp.233-246
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    • 2013
  • This study was performed to investigate the recognition of Korean farmers for the safety of environment-friendly organic materials (EFOMs). The main purchase line of environment-friendly organic materials was the sustainable agriculture organizations, and decisive basis for the materials purchase was because they thought that agricultural produce cultivated with EFOMs were safe. Currently farmers obtained the most information about EFOMs through farmer's education operated by agricultural technology center. Sixty nine percent of respondents hoped that they would get the information on the website and through official education opened by of the government or relative institutions. Whereas, only a few respondents hoped that they would get the information from the sellers. Therefore we thought that government and related institutions should operate the education program to give the sellers the scientific and accurate information about EFOMs. In addition, farmers are highly recognized the necessity of safety-related information provided by government and related institutions. Most Farmers thought that safety education for farmers using EFOMs was very important. Also, it was confirmed that farmer respondents thought to need to establish the shelf-life and user guideline and to label about toxic category of EFOMs. The Cronbach' alpha coefficient calculated in order to verify the reliability of survey questions was 0.699. That value indicates that responses of the respondents in this study might be reliable.

HPLC Analysis and Pharmacokinetics of KAL-1120, a Novel Anti-inflammation Agent, in Rats (흰쥐에서 항염증제 후보물질 KAL-1120의 HPLC 분석 및 약물동태)

  • Shin, Dae-Hwan;Lee, Jung-Yeol;Park, Seong-Hyeok;Lee, Gyeong-Bok;Han, Kun;Chung, Youn-Bok
    • Journal of Pharmaceutical Investigation
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    • v.40 no.2
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    • pp.101-107
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    • 2010
  • A rapid and sensitive reversed-phase high performance liquid chromatography (HPLC) method was developed for the determination of N-(-4-Chlorophenyl)-6-hydroxy-7-methoxy-2-chromanecarboxamide (KAL-1120), a novel anti-inflammation agent, in the rat plasma. The method was applied to analyze the compound in the biological fluids such as bile, urine and tissue homogenates. After liquid-liquid extraction, the compound was analyzed on an HPLC system with ultraviolet detection at 275 nm. HPLC was carried out using reversed-phase isocratic elution with a $C_{18}$ column, a mobile phase of a mixture of acetonitril (40 v/v%) at a flow rate of 1.0 mL/min. The chromatograms showed good resolution and sensitivity and no interference of plasma. The calibration curve for the drug in plasma was linear over the concentration range of 0.05-50 ${\mu}g$/mL. The intra- and inter-day assay accuracies of this method ranged from 0.06% to 9.33% of normal values and the precision did not exceed 6.28% of relative standard deviation. The plasma concentration of KAL-1120 decreased to below the quantifiable limit at 1.5 hr after the i.v. bolus administration of 2-10 mg/kg to rats ($t_{1/2,({\alpha})}$ and $t_{1/2,({\beta})$ of 2.15 and 26.7 min at a dose of 2 mg/kg, 3.91 and 33.0 min at a dose of 10 mg/kg, respectively). The steady-state volume of distribution ($V_{dss}$) and the total body clearance ($CL_t$) were not significantly altered in rats given doses from 2 to 10 mg/kg. Of the various tissues tested, KAL-1120 was mainly distributed in the lung and heart after i.v. bolus administration. KAL-1120 was detected in the bile by 30 min after its i.v. bolus administration. However, the concentration in the urine after i.v. bolus administration became too low to measure, suggesting that KAL-1120 is mostly excreted in the bile. In conclusion, this analytical method was suitable for the preclinical pharmacokinetic studies of KAL-1120 in rats.