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Selected papers from the latest issue:
Simple and rapid determination of thiol compounds by HPLC and fluorescence detection with 1,3,5,7-tetramethyl-8-phenyl- (2-maleimide) difluoroboradiaza-s-indacene
07 November 2011, 22:03:57
Publication year: 2011
Source: Journal of Chromatography B, Available online 7 November 2011
Xiao-Feng Guo, Pei-Xuan Zhao, Hong Wang, Hua-shan Zhang
A rapid and simple background-free high-performance liquid chromatographic (HPLC) approach has been developed for simultaneously determining free thiol compounds including coenzyme A (CoA), cysteine (Cys), glutathione (GSH) and N-acetyl-cysteine (NAC) in biological samples by using 1,3,5,7-tetramethyl-8-phenyl-(2-maleimide) difluoroboradiaza-s-indacene (TMPAB-o-M) as fluorogenic reagent. After derivatization under physiological conditions within 6 min, baseline separation was finished in just 6 min using isocratic elution with reversed-phase HPLC and fluorescence detection. Excellent linearity was observed for all analytes over their concentration ranges of 1-500 nM and detection limits ranging 0.13 nM for CoA to 0.25 nM for Cys (S/N = 3) were achieved. The utility of the proposed method has been validated by measuring thiol compounds mentioned above in tissue, fluid and cell samples. The results indicated that this approach was well suited for high-throughput quantitative determination of thiols and study of the physiological role of them.
Source: Journal of Chromatography B, Available online 7 November 2011
Xiao-Feng Guo, Pei-Xuan Zhao, Hong Wang, Hua-shan Zhang
A rapid and simple background-free high-performance liquid chromatographic (HPLC) approach has been developed for simultaneously determining free thiol compounds including coenzyme A (CoA), cysteine (Cys), glutathione (GSH) and N-acetyl-cysteine (NAC) in biological samples by using 1,3,5,7-tetramethyl-8-phenyl-(2-maleimide) difluoroboradiaza-s-indacene (TMPAB-o-M) as fluorogenic reagent. After derivatization under physiological conditions within 6 min, baseline separation was finished in just 6 min using isocratic elution with reversed-phase HPLC and fluorescence detection. Excellent linearity was observed for all analytes over their concentration ranges of 1-500 nM and detection limits ranging 0.13 nM for CoA to 0.25 nM for Cys (S/N = 3) were achieved. The utility of the proposed method has been validated by measuring thiol compounds mentioned above in tissue, fluid and cell samples. The results indicated that this approach was well suited for high-throughput quantitative determination of thiols and study of the physiological role of them.
Highlights
► We develop an HPLC-fluorescence method for analysis of thiol compounds. ► The derivatization of coenzyme A, Cys, GSH and N-acetyl-cysteine need only 6 min. ► Baseline separation is achieved in 6 min using isocratic elution. ► The chromatograms are background-free. ► The detection limits are from 0.13 nM for coenzyme A to 0.25 nM for Cys (S/N = 3).Simultaneous determination of codeine, ephedrine, guaiphenesin and chlorpheniramine in beagle dog plasma using high performance liquid chromatography coupled with tandem mass spectrometric detection: application to a bioequivalence study
07 November 2011, 22:03:57
Publication year: 2011
Source: Journal of Chromatography B, Available online 7 November 2011
Ziyan Hu, Qiaogen Zou, Jixin Tian, Lili Sun, Zunjian Zhang
A rapid and sensitive method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous determination of codeine, ephedrine, guaiphenesin and chlorpheniramine in beagle dog plasma has been developed and validated. Following liquid-liquid extraction, the analytes were separated on a reversed-phase C18column (150mm × 2.0 mm, 3 μm) using formic acid:10 mM ammonium acetate: methanol (0.2:62:38, v/v/v) as mobile phase at a flow rate of 0.2 mL/min and analyzed by a triple-quadrupole mass spectrometer in the selected reaction monitoring (SRM) mode. The method was linear for all analytes over the following concentration (ng/mL) ranges: codeine 0.08∼16; ephedrine 0.8∼160; guaiphenesin 80∼16000; chlorpheniramine 0.2∼40. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. It is the first time that the validated HPLC–MS/MS method was successfully applied to a bioequivalence study in 6 healthy beagle dogs.
Source: Journal of Chromatography B, Available online 7 November 2011
Ziyan Hu, Qiaogen Zou, Jixin Tian, Lili Sun, Zunjian Zhang
A rapid and sensitive method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous determination of codeine, ephedrine, guaiphenesin and chlorpheniramine in beagle dog plasma has been developed and validated. Following liquid-liquid extraction, the analytes were separated on a reversed-phase C18column (150mm × 2.0 mm, 3 μm) using formic acid:10 mM ammonium acetate: methanol (0.2:62:38, v/v/v) as mobile phase at a flow rate of 0.2 mL/min and analyzed by a triple-quadrupole mass spectrometer in the selected reaction monitoring (SRM) mode. The method was linear for all analytes over the following concentration (ng/mL) ranges: codeine 0.08∼16; ephedrine 0.8∼160; guaiphenesin 80∼16000; chlorpheniramine 0.2∼40. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. It is the first time that the validated HPLC–MS/MS method was successfully applied to a bioequivalence study in 6 healthy beagle dogs.
Highlights
► This is the first study to describe a sensitive and precise LC-MS/MS assay for codeine, ephedrine, guaiphenesin and chlorpheniramine. ► We utilized this method to measure concentration of codeine, ephedrine, guaiphenesin and chlorpheniramine in beagle dog plasma. ► This method supports research of codeine, ephedrine, guaiphenesin and chlorpheniramine and its pharmacokinetics in beagle dog plasma.Screening for in vitro metabolites ofAbelmoschus manihotextract in intestinal bacteria by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry
07 November 2011, 22:03:57
Publication year: 2011
Source: Journal of Chromatography B, Available online 7 November 2011
Caifu Xue, Shu Jiang, Jianming Guo, Dawei Qian, Jin-ao Duan, ...
Abelmoschus manihothas drawn much attention recently due to its potential beneficial health effects after oral administration. However, the metabolic fate ofAbelmoschus manihotin intestinal flora is not well understood. In this paper, we describe a strategy using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF MS) with automated data analysis software (MetaboLynx™) for fast analysis of the metabolic profile of flavonoids fromAbelmoschus manihotin intestinal flora. The human and rat incubated samples collected 72 h in the anaerobic incubator were analyzed by UPLC-Q-TOF MS within 10 min. A total of fourteen metabolites were identified in human and rat incubated solution compared with blank samples. The results indicated that hydrolysis, hydroxylation and acetylation were the major metabolic pathways of flavonoids inAbelmoschus manihotextract invitro. MSwas used for simultaneous acquisition of precursor ion information and fragment ion data at high and low collision energy in one analytical run, which facilitated the fast structural characterization of metabolites.This work demonstrated the potential of the UPLC-Q-TOF MS approach using Metabolynx for fast and automated identification of metabolites of natural product in intestinal flora.
Source: Journal of Chromatography B, Available online 7 November 2011
Caifu Xue, Shu Jiang, Jianming Guo, Dawei Qian, Jin-ao Duan, ...
Abelmoschus manihothas drawn much attention recently due to its potential beneficial health effects after oral administration. However, the metabolic fate ofAbelmoschus manihotin intestinal flora is not well understood. In this paper, we describe a strategy using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF MS) with automated data analysis software (MetaboLynx™) for fast analysis of the metabolic profile of flavonoids fromAbelmoschus manihotin intestinal flora. The human and rat incubated samples collected 72 h in the anaerobic incubator were analyzed by UPLC-Q-TOF MS within 10 min. A total of fourteen metabolites were identified in human and rat incubated solution compared with blank samples. The results indicated that hydrolysis, hydroxylation and acetylation were the major metabolic pathways of flavonoids inAbelmoschus manihotextract invitro. MSwas used for simultaneous acquisition of precursor ion information and fragment ion data at high and low collision energy in one analytical run, which facilitated the fast structural characterization of metabolites.This work demonstrated the potential of the UPLC-Q-TOF MS approach using Metabolynx for fast and automated identification of metabolites of natural product in intestinal flora.
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