• Title/Summary/Keyword: Liquid Chromatography

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Purification and Characterization of Very Low Density Lipoprotein in Commercial Broiler and Crossbred Village Chickens by Fast Protein Liquid Chromatography

  • Tan, B.K.;Foo, H.L.;Loh, Teck Chwen;Norhani, A.;Zulkifli, I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1780-1785
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    • 2005
  • Very low density lipoprotein (VLDL) of commercial broiler (CB) and crossbred village chicken (AK) was purified using Fast Protein Liquid Chromatography (FPLC). The fraction collected was then confirmed as VLDL using 4% polyacrylamide gel electrophoresis and transmission electron microscopy (TEM). The particle size of VLDL is 46.8${\pm}$8.6 nm. The VLDL fraction was then subfractionated and the apolipoprotein (apo) profile was studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDSPAGE). The CB and AK have almost similar types of apo in both subfractions 1 and 2. The AK showed the presence of apoAI, AIV, D and E whereas the CB had apo AIV, D, E and H. The apo AIV and apo E were present in both subfractions of AK and CB.

Antioxidant Activity Analysis of Useful Compounds from Artemisiae Annuae Herba Using On-line Screening HPLC-ABTS+ Assay (On-line Screening HPLC-ABTS+ assay를 이용한 청호로부터 유용성분의 항산화 활성 분석)

  • Lee, Kwang Jin;Ma, Jin Yeul
    • Journal of Applied Biological Chemistry
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    • v.57 no.4
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    • pp.301-305
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    • 2014
  • The Antioxidant activity screening identification of five kind compounds in Artemisiae annuae herba with the on-line screening high performance liquid chromatography (HPLC) $ABTS^+$ assay. The various experimental variables such as the extraction time (h) and extraction solvent composition (%) of dipping method were investigated efficiently extraction at the room temperature $25^{\circ}C$. The results, the highest yield of total extract amount (0.458 g, 15.250%) was obtained by dipping method with 100% water and extraction time to 3 h. And the on-line screening HPLC-$ABTS^+$ assay method was rapid and efficient to search for bioactivity from natural products.

Development of Analytical Method and Monitoring of Organophosphorus Pesticides in the Raw Water and Clean Water by Liquid Chromatography-Tandem Mass Spectrometry (LC/MS/MS를 이용한 유기인계 농약류의 최적 분석법 정립과 원·정수에서의 모니터링)

  • Kim, Gyung-A;Song, Mi-Jeong;Yeom, Hoon-Sik;Son, Hee-Jong;Lee, Sang-Won;Choi, Jin-Tack
    • Journal of Environmental Science International
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    • v.24 no.12
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    • pp.1569-1582
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    • 2015
  • The analytical method for 16 organophosphorus pesticides was developed in this study. The 16 organophosphorus pesticides were analyzed by liquid chromatography-tandem mass spectrometry (LC/MS/MS) using on-line solid phase extraction (on-line SPE) with PLRP- S cartridge. Analysis of all analytes in the MS/MS was processed in the electrospray ioni-zation (ESI) positive mode. They are Azinphos ethyl, Chlorfenvinphos, Ethion, Famphur, Phosmet, Phosphamidon, Terbufos, Aspon, Chlorpyrifos-methyl, Crotoxyphos, Dichlofenthi-on, Dicrotophos, Fonofos, Thionazin, Dimethoate and Iprobenfos. Limits of detection (LODs) and Limits of quantification(LOQs) were obtained as 0.8~2.0 ng/L and 2.6~6.4 ng/L, respectively. All compounds were not detected at the 8 sampling points of the raw water and clean water.

A Study on the Determination of Caffeine in Coffee, Black tea and Green Tea by high performance Liquid Chromatography (고속액체 크로마토크래피에 의한 커피, 홍차, 녹차중의 카페인 정량에 관한 연구)

  • 권익부;이윤수;우상규;이충영;서준걸
    • Journal of Food Hygiene and Safety
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    • v.5 no.4
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    • pp.213-217
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    • 1990
  • A simple and practical method for the determination of caffeine in coffee, black tea and green tea was studied. The analysis of caffeine was performed by reverse phase high perfomance liquid chromatography using a ${\mu}-Bondapak$ C18 column at isocratic condition with methanol-acetic acid-water (20: 1: 79) on UV detector at 280 nm. The extraction and clean-up of caffeine in sample is based on combing a simple pretreatment with the use of a Sep-Pak Alumina A cartridge. The average recoveries of caffeine from several samples were 95.2 -101.3%, the relative standard deviation for the whole procedure was 0.10 ~ 0.62%, and the detection limit of caffeine in sample solution was about $0.1\;\mu\textrm{g}\;per\;ml$.

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Development and Validation of a Liquid Chromatography-Tandem Mass Spectrometry Method for the Determination of ε-Acetamidocaproic Acid in Rat Plasma

  • Kim, Tae Hyun;Choi, Yong Seok;Choi, Young Hee;Kim, Yoon Gyoon
    • Toxicological Research
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    • v.29 no.3
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    • pp.203-209
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    • 2013
  • A simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of ${\varepsilon}$-acetamidocaproic acid (AACA), the primary metabolite of zinc acexamate (ZAC), in rat plasma by using normetanephrine as an internal standard. Sample preparation involved protein precipitation using methanol. Separation was achieved on a Gemini-NX $C_{18}$ column ($150mm{\times}2.0mm$, i.d., 3 ${\mu}m$ particle size) using a mixture of 0.1% formic acid-water : acetonitrile (80 : 20, v/v) as the mobile phase at a flow rate of 200 ${\mu}l/min$. Quantification was performed on a triple quadrupole mass spectrometer employing electrospray ionization and operating in multiple reaction monitoring (MRM) and positive ion mode. The total chromatographic run time was 4.0 min, and the calibration curves of AACA were linear over the concentration range of 20~5000 ng/ml in rat plasma. The coefficient of variation and relative error at four QC levels were ranged from 1.0% to 5.8% and from -8.4% to 6.6%, respectively. The present method was successfully applied for estimating the pharmacokinetic parameters of AACA following intravenous or oral administration of ZAC to rats.

In vitro Metabolism of Methallylescaline in Human Hepatocytes Using Liquid Chromatography-High Resolution Mass Spectrometry

  • Kim, Sunjoo;Kim, Ju-Hyun;Kim, Dong Kyun;Lee, Jaesin;In, Sangwhan;Lee, Hye Suk
    • Mass Spectrometry Letters
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    • v.9 no.3
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    • pp.86-90
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    • 2018
  • Methallylescaline, 2-(3,5-dimethoxy-4-[(2-methylprop-2-en-1-yl)oxy]phenyl)ethanamine, is a new psychoactive substance with potent agonist of 5-HT receptor, but there is little information on its pharmacological effect, metabolism, and toxicity. It is necessary to characterize the metabolic profiling of methallylescaline in human hepatocytes using liquid chromatography-high resolution mass spectrometry. Methallylescaline was metabolized to three hydroxy-methallylescaline (M1-M3) and dihydroxy-methallylescaline (M4) via hydroxylation in human hepatocytes. CYP2D6, CYP2J2, CYP1A2, and CYP3A4 enzymes were responsible for the metabolism of methallylescaline. The metabolites as well as methallylescaline would be used for monitoring the abuse of methallylescaline.

Metabolism and excretion of novel pulmonary-targeting docetaxel liposome in rabbits

  • Wang, Jie;Zhang, Li;Wang, Lijuan;Liu, Zhonghong;Yu, Yu
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.1
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    • pp.45-54
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    • 2017
  • Our study aims to determine the metabolism and excretion of novel pulmonary-targeting docetaxel liposome (DTX-LP) using the in vitro and in vivo animal experimental models. The metabolism and excretion of DTX-LP and intravenous DTX (DTX-IN) in New Zealand rabbits were determined with ultra-performance liquid chromatography tandem mass spectrometry. We found DTX-LP and DTX-IN were similarly degraded in vitro by liver homogenates and microsomes, but not metabolized by lung homogenates. Ultra-performance liquid chromatography tandem mass spectrometry identified two shared DTX metabolites. The unconfirmed metabolite $M_{un}$ differed structurally from all DTX metabolites identified to date. DTX-LP likewise had a similar in vivo metabolism to DTX-IN. Conversely, DTX-LP showed significantly diminished excretion in rabbit feces or urine, approximately halving the cumulative excretion rates compared to DTX-IN. Liposomal delivery of DTX did not alter the in vitro or in vivo drug metabolism. Delayed excretion of pulmonary-targeting DTX-LP may greatly enhance the therapeutic efficacy and reduce the systemic toxicity in the chemotherapy of non-small cell lung cancer. The identification of $M_{un}$ may further suggest an alternative species-specific metabolic pathway.

Liquid Chromatography-Tandem Mass Spectrometry Analysis of Riboflavin in Beagle Dog Plasma for Pharmacokinetic Studies

  • Jeong, Hyeon Myeong;Shin, Beom Soo;Shin, Soyoung
    • Mass Spectrometry Letters
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    • v.11 no.1
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    • pp.10-14
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    • 2020
  • Riboflavin is a water-soluble vitamin, which serves as a precursor to flavin mononucleotide and flavin adenine dinucleotide. This study aimed to develop a simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis for the quantification of riboflavin in the Beagle dog plasma. This method utilized simple protein precipitation with acetonitrile and 13C4, 15N2-riboflavin was used as an internal standard (IS). For chromatographic separation, a hydrophilic interaction liquid chromatography (HILIC) column was used with gradient elution. The mobile phase consisted of 0.1% (v/v) aqueous formic acid with 10 mM ammonium formate and acetonitrile with 0.1% (v/v) formic acid. Since riboflavin is an endogenous compound, 4% bovine serum albumin in phosphate buffered saline was used as a surrogate matrix to prepare the calibration curve. The quantification limit for riboflavin in the Beagle dog plasma was 5 ng/mL. The method was fully validated for its specificity, sensitivity, accuracy and precision, recovery, and stability according to the US FDA guidance. The developed LC-MS/MS method may be useful for the in vivo pharmacokinetic studies of riboflavin.

Determination of 11 Ginsenosides in Black Ginseng Developed from Panax ginseng by High Performance Liquid Chromatography

  • Sun, Bai-Shen;Gu, Li-Juan;Fang, Zhe-Ming;Wang, Chun-Yan;Wang, Zhen;Sung, Chang-Keun
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.561-564
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    • 2009
  • A high performance liquid chromatography (HPLC) method has been developed for determination of 11 ginsenosides in black ginseng (BG, white ginseng that is subjected to 9 cycles of $95^{\circ}C$ for 3 hr). After eluted by gradient elution of water-acetonitrile without buffer in 70 min, 11 ginsenosides in BG were identified. The proposed method provided good linearity ($R^2$>0.9995), accuracy (92.2-106.6%), and intra- and interday precision (RSD<2.6%). In addition, ginsenosides compositions in white, red, and black ginsengs were investigated using this method, respectively. Interestingly, in BG, the content of ginsenoside $Rg_3$ which does not existed in white ginseng was 7.51 mg/g, approximately 20 times than that in red ginseng.

Accurate Determination of Malachite Green and Leucomalachite Green in Fish using Isotope Dilution Liquid Chromatography/Mass Spectrometry (ID-LC/MS)

  • Ahn, Seong-Hee;Kim, Byung-Joo;Lee, Yun-Jung;Kim, Jeong-Kwon
    • Bulletin of the Korean Chemical Society
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    • v.31 no.11
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    • pp.3228-3232
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    • 2010
  • Malachite green (MG) has been used world-widely in aquaculture as a parasiticide or fungicide. Although MG performed successfully, it has not been permitted for use in aquaculture from European Union, USA, and Canada because of its carcinogenicity and mutagenicity. We developed a sensitive and specific method to determine MG and its principal metabolite, leucomalachite green (LMG), respectively by isotope dilution liquid chromatography mass spectrometry (ID-LC/MS). To enhance the extraction recovery of MG and LMG from fish tissue, an additional step, saponification, was introduced in sample preparation process to remove fat in sample extract, which hampered the performance of SPE columns. The residue of MG and LMG in fish was analyzed using liquid chromatography mass spectrometry in the selected ion monitoring (SIM) mode by monitoring at m/z 329 and 334 for MG and $d_5$-MG and at m/z 331 and 337 for LMG and $^{13}C_6$-LMG, respectively. This method was validated by comparing with the value of the reference material provided by Laboratory Government Chemistry (LGC). The results agreed within the measurement uncertainty and the accuracy was much improved than the provided reference value by LGC.