• Title/Summary/Keyword: Liquid Chromatography-tandem Mass Spectrometry

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A Study on the Degradability of Pharmaceuticals during UV Treatment (자외선 처리시의 의약품류의 분해도에 관한 연구)

  • Kim, Ilho
    • Journal of Korean Society on Water Environment
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    • v.28 no.6
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    • pp.902-910
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    • 2012
  • The photodegradation characteristics of 30 pharmaceuticals were investigated by batch experiments using Ultraviolet (UV) reactor. The investigated pharmaceuticals include antibiotics, analgesics and antiarrhythmic agents etc. Tested water was prepared by simultaneously spiking 30 pharmaceuticals into pure water, and each experiment was conducted using 3 types of UV lamps. As a result, batch experiments showed that reactions of all the investigated pharmaceuticals followed pseudo-first order reaction regardless of the applied UV lamps. Among the pharmaceuticals, Cyclophosphamide, 2-Quinoxaline carboxylic acid and Clarithromycin proved to be the most UV-resistant compounds. Contrarily, Ceftiofur, Diclofenac and Ketoprofen were easily degraded by all the UV lamps. Dissolved organic carbon (DOC) concentration hardly changed although the concentration of the pharmaceuticals concentration gradually decreased with time, indicating that the degradation of parent pharmaceuticals may produce their intermediates during UV treatment.

The Role of Sphingolipids Cycle in Hydrogen Peroxide-Induced Apoptosis in HL-60 Cells

  • Son , Jung-Hyun;Lee, Jae-Ick;Yang , Ryung;Kim, Dong-Hyun
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.213.1-213.1
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    • 2003
  • Sphingolipids and their metabolites are highly bioactive molecules that affect various cellular functions including differentiation, cellular senescence, apoptosis, and proliferation when added exogenously, or elevated intracellularly by turnover of complex sphingolipids or synthesis from de novo pathway. We are investigating the relationship of sphingolipids cycle in apoptosis early events. A new column liquid chromatography- tandem mass spectrometry (LC/MS/MS) in combination with multiple reaction monitoring (MRM) method was developed for the rapid, simultaneous and quantitative determination of unambiguous detecting sphingolipids in cells. (omitted)

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Identification of glycated peptides in human serum using LC-MS/MS: A comparison of data-dependent acquisition and Parallel reaction monitoring

  • Seonghyeon Cho;Yejin Jeon;Van-An Duong;Jong-Moon Park;Hookeun Lee
    • Analytical Science and Technology
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    • v.36 no.1
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    • pp.12-21
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    • 2023
  • Protein glycation is vital to aging and disease. However, glycated proteins are low-abundant in plasma, rendering them difficult to identify using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Many studies have analyzed glycated peptides with high reproducibility. Here, glycated peptides in human serum were analyzed by LC-MS/MS using data-dependent acquisition (DDA) and parallel reaction monitoring (PRM). Boronic acid (BA) enrichment of in vitro glycated human serum peptides was performed. BA enrichment identified the most glycated peptides, and the glycated peptides of the more diversified proteins, excluding albumin, were analyzed. In PRM, glycated albumin PSMs were the most common, and this method exhibited the best reproducibility. The results of this study could help compare methods for identifying glycation-related biomarkers.

Search for Novel Stress-responsive Protein Components Using a Yeast Mutant Lacking Two Cytosolic Hsp70 Genes, SSA1 and SSA2

  • Matsumoto, Rena;Rakwal, Randeep;Agrawal, Ganesh Kumar;Jung, Young-Ho;Jwa, Nam-Soo;Yonekura, Masami;Iwahashi, Hitoshi;Akama, Kuniko
    • Molecules and Cells
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    • v.21 no.3
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    • pp.381-388
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    • 2006
  • Heat shock proteins (Hsp) 70 are a ubiquitous family of molecular chaperones involved in many cellular processes. A yeast strain, ssa1/2, with two functionally redundant cytosolic Hsp70s (SSA1 and SSA2) deleted shows thermotolerance comparable to mildly heatshocked wild type yeast, as well as increased protein synthesis and ubiquitin-proteasome protein degradation. Since mRNA abundance does not always correlate well with protein expression levels it is essential to study proteins directly. We used a gel-based approach to identify stress-responsive proteins in the ssa1/2 mutant and identified 43 differentially expressed spots. These were trypsin-digested and analyzed by nano electrospray ionization liquid chromatography tandem mass spectrometry (nESI-LC-MS/MS). A total of 22 non-redundant proteins were identified, 11 of which were confirmed by N-terminal sequencing. Nine proteins, most of which were up-regulated (2-fold or more) in the ssa1/2 mutant, proved to be stress-inducible proteins such as molecular chaperones and anti-oxidant proteins, or proteins related to carbohydrate metabolism. Interestingly, a translational factor Hyp2p up-regulated in the mutant was also found to be highly phosphorylated. These results indicate that the cytosolic Hsp70s, Ssa1p and Ssa2p, regulate an abundance of proteins mainly involved in stress responses and protein synthesis.

In Vitro Inhibitory Effect of Licoricidin on Human Cytochrome P450s

  • Kim, Sunju;O, Heungchan;Kim, Jeong Ah;Lee, Seung Ho;Lee, Sangkyu
    • Mass Spectrometry Letters
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    • v.5 no.3
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    • pp.84-88
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    • 2014
  • Licoricidin isolated from Glycyrrhiza uralensis is known to have anticancer, anti-nephritic, anti-Helicobacter pylori, and antibacterial effects. In this study, a cocktail probe assay and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to investigate the modulating effect of licoricidin on cytochrome P450 (CYP) enzymes in human liver microsomes. When licoricidin was incubated at $0-25{\mu}m$ with CYP probes for 60 min at $37^{\circ}C$, it showed potent inhibitory effects on CYP2B6-catalyzed bupropion hydroxylation and CYP2C9-catalyzed diclofenac 4'-hydroxylation with half maximal inhibitory concentration ($IC_{50}$) values of 3.4 and $4.0{\mu}m$, respectively. The inhibition mode of licoricidin was revealed as competitive, dose-dependent, and non-time-dependent, and following the pattern of Lineweaver-Burk plots. The inhibitory effect of licoricidin has been confirmed in human recombinant cDNA-expressed CYP2B6 and 2C9 with $IC_{50}$ values of 4.5 and $0.73{\mu}m$, respectively. In conclusion, this study has shown the potent inhibitory effect of licoricidin on CYP2B6 and CYP2C9 activity could be important for predicting potential herb-drug interactions with substrates that mainly undergo CYP2B- and CYP2C9-mediated metabolism.

Correlation between the Content and Pharmacokinetics of Ginsenosides from Four Different Preparation of Panax Ginseng C.A. Meyer in Rats

  • Jeon, Ji-Hyeon;Lee, Jaehyeok;Lee, Chul Haeng;Choi, Min-Koo;Song, Im-Sook
    • Mass Spectrometry Letters
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    • v.12 no.1
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    • pp.16-20
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    • 2021
  • We aimed to compare the content of ginsenosides and the pharmacokinetics after the oral administration of four different ginseng products at a dose of 1 g/kg in rats. The four different ginseng products were fresh ginseng extract, red ginseng extract, white ginseng extract, and saponin enriched white ginseng extract prepared from the radix of Panax ginseng C.A. Meyer. The ginsenoside concentrations in the ginseng product and the rat plasma samples were determined using a liquid chromatography-tandem mass spectrometry (LC-MS/MS). Eight or nine ginsenosides of the 15 tested ginsenosides were detected; however, the content and total ginsenosides varied depending on the preparation method. Moreover, the content of triglycosylated ginsenosides was higher than that of diglycosylated ginsenosides, and deglycosylated ginsenosides were not present in any preparation. After the single oral administrations of four different ginseng products in rats, only four ginsenosides, such as 20(S)-ginsenosides Rb1 (GRb1), GRb2, GRc, and GRd, were detected in the rat plasma samples among the 15 ginsenosides tested. The plasma concentrations of GRb1, GRb2, GRc, and GRd were different depends on the preparation method but pharmacokinetic features of the four ginseng products were similar. In conclusion, a good correlation between the area under the concentration curve and the content of GRb1, GRb2, and GRc, but not GRd, in the ginseng products was identified and it might be the result of their higher content and intestinal biotransformation of the ginseng product.

Purification and Identification of Antioxidant Peptides from Enzymatic Hydrolysate of Spirulina platensis

  • Yu, Jie;Hu, Yuanliang;Xue, Mingxiong;Dun, Yaohao;Li, Shenao;Peng, Nan;Liang, Yunxiang;Zhao, Shumao
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1216-1223
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    • 2016
  • The aim of this study was to isolate antioxidant peptides from an enzymatic hydrolysate of Spirulina platensis. A novel antioxidant peptide was obtained by ultrafiltration, gel filtration chromatography, and reverse-phase high-performance liquid chromatography, with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay used to measure the antioxidant activity, and the sequence was determined to be Pro-Asn-Asn (343.15 Da) by electrospray ionization tandem mass spectrometry. This peptide was synthesized to confirm its antioxidant properties, and it exhibited 81.44 ± 0.43% DPPH scavenging activity at 100 μg/ml, which was similar to that of glutathione (82.63 ± 0.56%). Furthermore, the superoxide anion and hydroxyl free-radical scavenging activities and the SOD activity of the peptide were 47.84 ± 0.49%, 54.01 ± 0.82%, and 12.55 ± 0.75%, respectively, at 10 mg/ml. These results indicate that S. platensis is a good source of antioxidant peptides, and that its hydrolysate may have important applications in the pharmaceutical and food industries.

Analytical method study for cephalexin with high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) applicable for residue studies in the whiteleg shrimp Litopenaeus vannamei (흰다리새우(Litopenaeus vannamei)에서 cephalexin의 잔류농도 연구를 위한 LC-MS/MS 분석법 연구)

  • Yang, Chan Yeong;Bae, Jun Sung;Lee, Chae Won;Jeong, Eun Ha;Lee, Ji-Hoon;Bak, Su-Jin;Choi, Sang-Hoon;Park, Kwan Ha
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.71-80
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    • 2021
  • Cephalexin, a semi-synthetic cephalosporin antibiotic, has long been used in fish aquaculture in various countries under legal authorization. The drug is thus widely available for use in other aquatic species except fishes like the crustacean whiteleg shrimp. This study aims to develop a sensitive method for laboratory residue studies to adopt in withdrawal period determinations. Through repeated trials from the existing methods developed for other food animal tissues, it was possible to achieve a sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method. The results showed that at a concentration of 0.1 mg/kg, the recovery rate was 81.79%, and C.V. value was 8.2%, which meet the recovery rate and C.V. recommended by Codex guideline. After satisfactory validation of analytical procedures, applicability to the shrimp tissue was confirmed in experimentally cephalexin-treated whiteleg shrimp. As a result, most muscle samples were detected below the limit of quantification (0.05 mg/kg) after day 3, and most hepatopancreas samples were detected below the limit of quantification after day 14. In particular, the limit of quantification 0.05 ppm with the presently developed method suggests sufficient sensitive over the current legal maximum residue limit of 0.2 mg/kg set for fishes.

Bioequivalence of LesacinTM Tablet to Jeil CravitTM Tablet (Levofloxacin 100 mg) by Liquid Chromatography- Electrospray Tandem Mass Spectrometry (LC-MS/MS를 이용한 제일크라비트정(레보플록사신 100 mg)에 대한 레사신정 100 mg의 생물학적 동등성)

  • Lee, Jin-Sung;Choi, Sang-Jun;Ryu, Ju-Hee;Seo, Ji-Hyung;Lee, Myung-Jae;Kang, Jong-Min;Tak, Sung-Kwon;Kang, Jin-Yang;Lee, Kyung-Tae
    • Journal of Pharmaceutical Investigation
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    • v.38 no.4
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    • pp.269-275
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    • 2008
  • The purpose of the present study was to evaluate the bioequivalence of two levofloxacin tablets, Jeil $Cravit^{TM}$ tablet (Jeil Pharm. Co., Ltd., Korea, reference drug) and $Lesacin^{TM}$ tablet (Ilhwa. Co., Ltd., Korea, test drug), according to the guidelines of Korea Food and Drug Administration (KFDA). Twenty-four healthy male Korean volunteers received two tablets containing levofloxacin 200 mg in a $2{\times}2$ crossover study. There was a one-week washout period between the doses. Plasma concentrations of levofloxacin were monitored for over a period of 24 hr after administration by using a high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). The area under the plasma concentration-time curve from time zero to 24 hr ($AUC_t$), maximum plasma drug concentration ($C_{max}$) and time to reach $C_{max}\;(T_{max})$ were complied from the plasma concentration-time data. Analysis of variance (ANOVA) test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$ and $C_{max}$. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for $Lesacin^{TM}$/Jeil $Cravit^{TM}$ were $\log\;0.9527{\sim}\log\;0.9981$ and $\log\;0.8712{\sim}\log\;1.0556$, respectively. These values were within the acceptable bioequivalence intervals of $\log\;0.80{\sim}\log\;1.25$, recommended by KFDA. In all of these results, we concluded that $Lesacin^{TM}$ tablet was bioequivalent to Jeil $Cravit^{TM}$ tablet, in terms of rate and extent of absorption.

Simultaneous determination of preservatives in sausages using liquid chromatography with electrospray ionization tandem mass spectrometry (LC-MS/MS를 이용한 소시지 중 보존료 동시분석)

  • Koh, Ba-Ra-Da;Kim, Ji-Yeon;Jang, Mi-Sun;Seo, Doo-Ri;Jung, Bo-Ram;Shin, Ji-Hyun;Lim, Jin-Taek;Kim, Yong-Hwan;Kim, Eun-Sun
    • Korean Journal of Veterinary Service
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    • v.38 no.2
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    • pp.127-136
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    • 2015
  • A simultaneous determination method was developed for nine preservatives (benzoic acid, sorbic acid, dehydroacetic acid, methyl-, ethyl-, isopropyl-, propyl-, isobutyl- and butyl-parabens) in sausage by liquid chromatography with electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS). Each parameter was established by multiple reaction monitoring in negative mode. Separation was achieved on a phenyl-hexyl ($2.5{\mu}m$, $2.1{\times}150mm$, Waters) with A-20 mM ammonium acetate containing 0.1% acetic acid in water, B-Acetonitrile as mobile phase with gradient mode at a flow rate of 0.3 mL/min. The developed method was validated for specificity, linearity, accuracy and precision in sausages samples. Linearity was over 0.998 with calibration curve of the mixed standards. The mean recoveries from sausages fortified at the level of 2.0~10.0 mg/L were in range of 98.60~109.16% with RSDs lower than 8.93%. The limits of detection (LOD) and the limits of quantification (LOQ) were in the range between 0.0003~0.085 mg/L and 0.01~0.257 mg/L, respectively. Intra-day precision and inter-day precision were 0.45~6.16% and 2.81~13.33%, respectively. Using presently developed determination method, 33 field sausage samples from Gwangju city in Korea were screened over nine preservatives. As a result, no preservatives were detected in all samples.