• Title/Summary/Keyword: LC-MS-MS

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Accumulation of Microcystins in Fish and Evaluation of Potential Human Health Risks: A Case Study on a Eutrophic Reservoir in Korea (마이크로시스틴의 어류내 축적성 및 인체 위해성 평가: 국내 저수지 사례연구)

  • Yoon, Hyojung;Seo, Jungkwan;Kim, Taksoo;Jo, Areum;Kim, Jungkon;Lee, Doohee;Kim, Pilje;Choi, Kyunghee
    • Journal of Environmental Health Sciences
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    • v.42 no.1
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    • pp.10-18
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    • 2016
  • Objectives: Microcystin (MC) produced during cyanobacterial blooms is a worldwide problem presenting a serious health threats to humans and ecosystems. During July through October of 2013, the Ilwol Reservoir experienced a high biomass of phytoplankton (maximum $211.7mg/m^3$ of Chlorophyll-a) containing the toxigenic cyanobacterium Oscillatoria sp. The aim of this study is to analyze MC concentration in the reservoir water, as well as in representative fish species (Carassius cuvieri, Carassius auratus, Channa argus). We also evaluated the human health risk of exposure to MCs accumulated in the fish. Methods: Concentrations of MCs in the water and fish samples were analyzed by liquid chromatography with a triple quadrupole tandem mass spectrometer (LC/MS/MS) and enzyme-linked immunosorbent assay (ELISA). Results: The total levels of four MC variants, including MC-LR, MC-RR, MC-YR and MC-LA were below the WHO drinking water guideline limit (1 ug MC-LR per liter) both for the dissolved and particulate fraction present in the water samples. The mean MC concentrations in the livers of all species were significantly higher than in the gills (p < 0.01) and muscles (p < 0.05). The values of estimated daily intake of MCs in muscles, the edible part of the fish, would be only $0.005-0.015{\mu}g/kg{\cdot}day$, much lower than WHO's provisional tolerable daily intake of $0.04{\mu}g/kg{\cdot}day$. Conclusion: This study suggests that, owing to the spatial distribution or temporal variation of MC, there is a need for careful monitoring of cyanotoxin in reservoir water and aquatic animals to protect public health.

Effect of Gamiondam-tang (GMODT), a Polyherbal Formula on the Pharmacokinetics Profiles of Tamoxifen in Male SD Rats

  • Ryu, Eun-A;Kang, Su-Jin;Song, Chang-Hyun;Lee, Bong-Hyo;Choi, Seong-Hun;Han, Chang-Hyun;Lee, Young-Joon;Ku, Sae-Kwang
    • The Journal of Korean Medicine
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    • v.38 no.2
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    • pp.61-72
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    • 2017
  • Objectives: The effects of Gamiondam-tang (GMODT) co-administration within 5min on the pharmacokinetics (PK) of tamoxifen were observed as a process of the comprehensive and integrative medicine, combination therapy of tamoxifen with GMODT to achieve synergic pharmacodynamics and reduce toxicity on the breast cancer. Methods: After 50mg/kg of tamoxifen treatment, GMODT 100mg/kg was administered within 5min. The plasma were collected at 30 min before administration, 30 min, 1, 2, 3, 4, 6, 8 and 24 hrs after end of GMODT treatment, and plasma concentrations of tamoxifen were analyzed using LC-MS/MS methods. PK parameters of tamoxifen (Tmax, Cmax, AUC, $t_{1/2}$ and $MRT_{inf}$) were analysis as compared with tamoxifen single administered rats using noncompartmental pharmacokinetics data analyzer programs. Results: Co-administration with GMODT induced increased trends of plasma tamoxifen concentrations to 1hr after end of administration, and then showed decreased trends of plasma tamoxifen concentrations, and especially significant (p<0.05) increases of plasma tamoxifen concentrations were demonstrated at 0.5hr after end of co-administration with GMODT and also related significant (p<0.05) decreases of $AUC_{0-inf}$ and $MRT_{inf}$ as compared with tamoxifen single formula treated rats, at dosage levels of tamoxifen 10 mg/kg and GMODT 100 mg/kg within 5 min, in this experiment. Conclusion: Based on the results of the present study, it is considered that single co-administration GMODT within 5min significantly inhibited the oral bioavailability of tamoxifen through variable influences on the absorption and excretion of tamoxifen, can be influenced on the toxicity or pharmacodynamic of tamoxifen.

Component Analysis and Anti-Proliferative Effects of Ethanol Extracts of Fruits, Leaves, and Stems from Elaeagnus umbellata in HepG2 Cells (보리수나무 열매, 잎 및 줄기 에탄올 추출물의 함유성분 분석과 간암 세포 증식억제 효과)

  • Kim, Min-Ju;Lim, Jong-Soon;Yang, Seun-Ah
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.6
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    • pp.828-834
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    • 2016
  • The aim of this study was to evaluate the physicochemical properties and antioxidant and anti-proliferative activities of different plant parts of Elaeagnus umbellata Thunb. extracted with ethanol (EtOH). EtOH extract of stems presenting the highest content of polyphenols showed the strongest 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity ($EC_{50}=54.04{\mu}g/mL$). The total content of free amino acids decreased in the order of leaves (6,179.12 mg/100 g)> stems (1,211.69 mg/100 g)> fruits EtOH extract (378.88 mg/100 g), and asparagine (1,907.57 mg/100 g), ${\gamma}-aminobutyric$ acid (300.17 mg/100 g), and proline (233.48 mg/100 g) were the major free amino acid in leaves, stems, and fruits, respectively. Five phenolic compounds in each extract were measured by using liquid chromatography- tandem mass spectrometry, and gallic acid (98.95 mg/100 g) and (+)-catechin (1,575.99 mg/100 g) were present as major components in leaves and stems, respectively. EtOH extract of leaves showed the highest anti-proliferative activity against HepG2 cells as measured by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazoliumbromide and lactate dehydrogenase assay but had no effects on Chang liver cells.

Optimal Conditions for Anthocyanin Extraction from Black Rice Bran and Storage Stability of Anthocyanin Extract (흑미 미강으로부터 안토시아닌의 최적 추출 조건 및 안토시아닌 추출 분말의 저장 안정성)

  • Kim, Hyo Ju;Wee, Ji-Hyang;Yang, Eun Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.10
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    • pp.1543-1549
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    • 2015
  • Black rice bran, a by-product from rice milling process, is a good natural source of anthocyanin pigment. The purpose of this study was to determine the optimum conditions for anthocyanin extraction from black rice bran as well as the stability of anthocyanin extract according to different storage temperatures. The main anthocyanin in 'Heugkwang' rice bran was identified as cyanidine-3-glucoside (C3G) by HPLC and LC-MS/MS. The yield and C3G content of black rice bran extract were investigated with various extraction solvents, temperatures, and times. The results indicate that the optimum extraction solvent, temperature, and time were 50% ethanol, $70^{\circ}C$, and 2 h, respectively. The stability of anthocyanin extract was studied during a storage period of 168 days at various temperatures ($-20^{\circ}C$, $4^{\circ}C$, and room temperature). Hunter's values (L, a, and b) of anthocyanin extract increased, whereas C3G content continuously decreased up to 168 days. Variations in Hunter's values and C3G content become larger as storage temperature increased. Anthocyanin extract from black rice bran was very stable, as C3G content after storage at all temperatures was maintained at more than 90% of initial content. These results suggest that anthocyanin extract from black rice bran may be useful as a natural food colorant.

Differential Cytotoxic Effects of Jaspine B in Various Cancer Cells (다양한 암세포주에서 Jaspine B의 함암활성 비교)

  • Lee, Jihoon;Choi, Kwangik;Kwon, Mihwa;Lee, Dongjoo;Choi, Min-Koo;Song, Im-Sook
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1392-1399
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    • 2016
  • Jaspine B is an anhydrophytosphingosine that is isolated from a marine sponge. Because of its structural similarity to sphingosine, it shows anti-cancer effects in human carcinomas. Therefore, this study aims to investigate its anti-proliferative effect on various cancer cells and to correlate its association with the intracellular accumulation of Jaspine B in relevant cancer cells. The anti-proliferative effect of Jaspine B in various cancer cells was determined by a cell viability test, and the intracellular concentration of Jaspine B in relevant cancer cells was determined using mass spectrometry coupled with liquid chromatography. The correlation coefficient and p value between the cytotoxicity and the cell accumulation of Jaspine B were determined using SPSS 16.1. The cytotoxicity of Jaspine B varied depending on the type of cancer cell when compared the $EC_{50}$ values of Jaspine B. Breast and melanoma cancer cells were susceptible to Jaspine B, whereas renal carcinoma cells were resistant. The intracellular concentrations of Jaspine B had a reciprocal correlation with the $EC_{50}$ values in the same cells (r = 0.838). The results suggested that the anti-proliferative effect of Jaspine B was associated with the cellular accumulation of this compound. However, Jaspine B was not a substrate for P-glycoprotein and breast cancer resistance protein, as major efflux pumps caused multidrug resistance. The maintenance of a high intracellular concentration is crucial for the cytotoxic effect of Jaspine B; however, efflux pumps may not be a controlling factor for Jaspine B-related resistance in cancer cells.

Analysis of Medroxyprogesterone Acetate in Meats (유통 식육 중 성장호르몬제 초산메드록시프로게스테론의 분석)

  • Lee, Yoon-Ae;Shim, Jee-Youn;Lee, Ryun-Kyung;Kim, Seung-Hwan;Oh, Hyun-Sook;Cho, Tae-Yong
    • Journal of Food Hygiene and Safety
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    • v.25 no.3
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    • pp.226-231
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    • 2010
  • We aimed at the monitoring medroxyprogesterone acetate (MPA) residue amount in meats and confirmed the safety of its residue in meats. Optimized condition for analytical and instrumental methods was obtained by method validation. The limit of detection (LOD) and limit of quantification (LOQ) were validated at 1.5 and 5.0 ug/kg, respectively. The calibration curve showed good linearity ($r^2$ = 0.9968) within the concentration range of 5.0~50.0 ug/kg. We selected progesterone-d9 for internal standard, The recoveries in fortified meat ranged from 67.5 to 109.56% at the 3 spiking levels. As the regulation of MPA analysis method used by LC-MS/MS on other products have established. We selected 3 species of farm stock products (cattle, pig, chicken) and purchased at the markets of seven major cities. The total 196 of meat including 46 of domestic beef, 43 of import beef, 60 of domestic pork, 12 of import pork and 35 of domestic chicken. No residue of synthetic growth hormones were detected in cattle, pig and chicken samples tested.

Application and Validation of an Optimal Analytical Method using QuEChERS for the determination of Tolpyralate in Agricultural Products (QuEChERS법을 활용한 농산물 중 제초제 Tolpyralate의 최적 분석법 선발 및 검증)

  • Lee, Han Sol;Park, Ji-Su;Lee, Su Jung;Shin, Hye-Sun;Kim, Ji-Young;Yun, Sang Soon;Jung, Yong-hyun;Oh, Jae-Ho
    • Korean Journal of Environmental Agriculture
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    • v.39 no.3
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    • pp.246-252
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    • 2020
  • BACKGROUND: Pesticides are broadly used to control weeds and pests, and the residues remaining in crops are managed in accordance with the MRLs (maximum residue limits). Therefore, an analytical method is required to quantify the residues, and we conducted a series of analyses to select and validate the quick and simple analytical method for tolpyralate in five agricultural products using QuEChERS (quick, easy, cheap, effective, rugged and safe) method and LC-MS/MS (liquid chromatography-tandem mass spectrometry). METHODS AND RESULTS: The agricultural samples were extracted with acetonitrile followed by addition of anhydrous magnesium sulfate, sodium chloride, disodium hydrogencitrate sesquihydrate and trisodium citrate dihydrate. After shaking and centrifugation, purification was performed with d-SPE (dispersive-solid phase extraction) sorbents. To validate the optimized method, its selectivity, linearity, LOD (limit of detection), LOQ (limit of quantitation), accuracy, repeatability, and reproducibility from the inter-laboratory analyses were considered. LOQ of the analytical method was 0.01 mg/kg at five agricultural products and the linearity of matrix-matched calibration were good at seven concentration levels, from 0.0025 to 0.25 mg/L (R2≥0.9980). Mean recoveries at three spiking levels (n=5) were in the range of 85.2~112.4% with associated relative standard deviation values less than 6.2%, and the coefficient of variation between the two laboratories was also below 13%. All optimized results were validated according to the criteria ranges requested in the Codex Alimentarius Commission (CAC) and Ministry of Food and Drug Safety (MFDS) guidelines. CONCLUSION: In conclusion, we suggest that the selected and validated method could serve as a basic data for detecting tolpyralate residue in imported and domestic agricultural products.

Prevalence of Detection of Prohibited Drugs on Doping Tests of Pre- and Post-races in Korea (2002-2013) (국내 경주마 도핑검사에서 금지약물 검출빈도 및 종류(2002-2013))

  • Yang, Jaehyuk;Han, Kwan-Seok;Yang, Young-Jin;Lim, Yoon-Kyu
    • Journal of Veterinary Clinics
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    • v.32 no.2
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    • pp.215-217
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    • 2015
  • There was no study on the prevalence of doping control of racehorses in Korea. The purpose of this study was to determine prohibited substances in horse races of a drugs testing program. Blood samples were taken from those 298,543 starters prior to racing and the that finished top 3 runners of each race and horses designated by the stewards shall be taken for collection of 91,482 urine samples for the purpose of post-race doping test in Seoul, Busan and Jeju Race Park between 2002 and 2013. Detection and measurement of prohibited substances were carried out by ELISA, GC/MS and LC/MS using standard methods at the Doping Control Center, Korea Racing Authority. Total 0.0030% of pre-races and total 0.0186% of post-races tested positive for prohibited substances. In pre- and post-race, caffeine and ketoprofen were the most detected prohibited substance respectively. We thought that characteristics of pharmacokinetics and pharmacodynamics of drugs resulted in different between plasma and urine. These substances have also been detected with other prohibited drugs suggesting that unintentional feeding or bedding may be the reason and groomers' confusion of candidate horses based by the stewards' research.

Effect of Gamiondam-tang (GMODT), a Polyherbal Formula on the Pharmacokinetics Profiles of Tamoxifen in Male SD Rats (2) - Single Oral Combination Treatment of Tamoxifen 50 mg/kg with GMODT 100 mg/kg with 2.5 hr-intervals -

  • Ryu, Eun-A;Kang, Su-Jin;Song, Chang-Hyun;Lee, Bong-Hyo;Choi, Seong-Hun;Han, Chang-Hyun;Lee, Young-Joon;Ku, Sae-Kwang
    • Journal of Society of Preventive Korean Medicine
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    • v.21 no.2
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    • pp.127-137
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    • 2017
  • Objectives : In our previous study, single co-administration GMODT within 5 min significantly inhibited the oral bioavailability of tamoxifen through variable influences on the absorption and excretion of tamoxifen. Therefore, the object of this study was to elucidate the possible effects on the pharmacokinetics of tamoxifen after single oral co-administration of GMODT with 2.5 hr-intervals. Methods : After 50 mg/kg of tamoxifen treatment, GMODT 100 mg/kg was administered with 2.5 hr-intervals. The plasma were collected at 30 min before administration, 30 min, 1, 2, 3, 4, 6, 8 and 24 hrs after end of GMODT treatment, and plasma concentrations of tamoxifen were analyzed using LC-MS/MS methods. PK parameters of tamoxifen (Tmax, Cmax, AUC, $t_{1/2}$ and $MRT_{inf}$) were analysis as compared with tamoxifen single administered rats. Results : Two-half hr-interval co-administration with GMODT induced variable changes on the plasma tamoxifen concentrations as compared with tamoxifen single treated rats, and especially significant (p<0.05) increases of plasma tamoxifen concentrations were demonstrated at 0.5 (199.61%) and 1 hr (101.06%) after end of co-administration with GMODT, and also related significant (p<0.05) decreases of $t_{1/2}$ (-39.54%) and $MRT_{inf}$ (-43.94%) as compared with tamoxifen single formula treated rats, at dosage levels of tamoxifen 50 mg/kg and GMODT 100 mg/kg with 2.5 hr-intervals, in this experiment. Conclusions : According to the results, GMODT critically decreased on the oral bioavailability of tamoxifen through variable influences on the absorption and excretion of tamoxifen. Hence, the co-administration of GMODT and tamoxifen should be avoided in the comprehensive and integrative medicine, combination therapy of tamoxifen with GMODT on the breast cancer.

Measurement of Human Cytochrome P450 Enzyme Induction Based on Mesalazine and Mosapride Citrate Treatments Using a Luminescent Assay

  • Kim, Young-Hoon;Bae, Young-Ji;Kim, Hyung Soo;Cha, Hey-Jin;Yun, Jae-Suk;Shin, Ji-Soon;Seong, Won-Keun;Lee, Yong-Moon;Han, Kyoung-Moon
    • Biomolecules & Therapeutics
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    • v.23 no.5
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    • pp.486-492
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    • 2015
  • Drug metabolism mostly occurs in the liver. Cytochrome P450 (CYP) is a drug-metabolizing enzyme that is responsible for many important drug metabolism reactions. Recently, the US FDA and EU EMA have suggested that CYP enzyme induction can be measured by both enzymatic activity and mRNA expression. However, these experiments are time-consuming and their interassay variability can lead to misinterpretations of the results. To resolve these problems and establish a more powerful method to measure CYP induction, we determined CYP induction by using luminescent assay. Luminescent CYP assays link CYP enzyme activity to firefly luciferase luminescence technology. In this study, we measured the induction of CYP isozymes (1A2, 2B6, 2C9, and 3A4) in cryopreserved human hepatocytes (HMC424, 478, and 493) using a luminometer. We then examined the potential induction abilities (unknown so far) of mesalazine, a drug for colitis, and mosapride citrate, which is used as an antispasmodic drug. The results showed that mesalazine promotes CYP2B6 and 3A4 activities, while mosapride citrate promotes CYP1A2, 2B6, and 3A4 activities. Luminescent CYP assays offer rapid and safe advantages over LC-MS/MS and qRT-PCR methods. Furthermore, luminescent CYP assays decrease the interference between the optical properties of the test compound and the CYP substrates. Therefore, luminescent CYP assays are less labor intensive, rapid, and can be used as robust tools for high-throughput CYP screening during early drug discovery.