Quantification of Acetaminophen and Its Metabolites in Plasma Using UPLC-MS: Doors Open to Therapeutic Drug Monitoring in Special Patient Populations

Item does not contain fulltextBACKGROUND: Acetaminophen (APAP, paracetamol) is the most commonly used drug for pain and fever in both the United States and Europe and is considered safe when used at registered dosages. Nevertheless, differences between specific populations lead to remarkable changes in exposure to potentially toxic metabolites. Furthermore, extended knowledge is required on metabolite formation after intoxication, to optimize antidote treatment. Therefore, the authors aimed to develop and validate a quick and easy analytical method for simultaneous quantification of APAP, APAP-glucuronide, APAP-sulfate, APAP-cysteine, APAP-glutathione, APAP-mercapturate, and protein-derived APAP-cysteine in human plasma by ultraperformance liquid chromatography-electrospray ionization-tandem mass spectrometry. METHODS: The internal standard was APAP-D4 for all analytes. Chromatographic separation was achieved with a reversed-phase Acquity ultraperformance liquid chromatography HSS T3 column with a runtime of only 4.5 minutes per injected sample. Gradient elution was performed with a mobile phase consisting of ammonium acetate, formic acid in Milli-Q ultrapure water or in methanol at flow rate of 0.4 mL/minute. RESULTS: A plasma volume of only 10 muL was required to achieve both adequate accuracy and precision. Calibration curves of all 6 analytes were linear. All analytes were stable for at least 48 hours in the autosampler; the high quality control of APAP-glutathione was stable for 24 hours. The method was validated according to the U.S. Food and Drug Administration guidelines. CONCLUSIONS: This method allows quantification of APAP and 6 metabolites, which serves purposes for research, as well as therapeutic drug monitoring. The advantage of this method is the combination of minimal injection volume, a short runtime, an easy sample preparation method, and the ability to quantify APAP and all 6 metabolites

Nocapyrones A−D, γ-Pyrones from aNocardiopsisStrain Isolated from the Marine SpongeHalichondria panicea

Four new γ-pyrones, nocapyrones A−D (1−4), were isolated from an organic extract of the Nocardiopsis strain HB383, which was isolated from the marine sponge Halichondria panicea. These are the first γ-pyrones reported from a Nocardiopsis strain. The structures were elucidated on the basis of one- and two-dimensional NMR experiments and supported by HPLC-UV/MS and HRESIMS analyses. The biosynthesis of nocapyrone A was investigated by feeding experiments with 13C-labeled compounds. In addition, one diketopiperazine, which was only known as a synthetic compound before, was isolated. The bioactivies of 1, 2, and the diketopiperazine were evaluated in a panel of assays

Mayamycin, a Cytotoxic Polyketide from aStreptomycesStrain Isolated from the Marine SpongeHalichondria panicea

A new benz[a]anthracene derivative called mayamycin (1) was identified in cultures of Streptomyces sp. strain HB202, which was isolated from the marine sponge Halichondria panicea and selected because of its profound antibiotic activity. The ability to produce aromatic polyketides was indicated by genetic analyses, demonstrating the presence of a type II polyketide synthase. The production of mayamycin (1) was induced by variation of the culture conditions. The structure of 1 was elucidated by HPLC-UV/MS and NMR spectroscopy. Mayamycin (1) exhibited potent cytotoxic activity against eight human cancer cell lines and showed activity against several bacteria including antibiotic-resistant strains

N-cycling and balancing of the N-deficit generated in the oxygen minimum zone over the Namibian shelf-An isotope-based approach

The northern Benguela upwelling system is a nutrient‐replete region with high plankton biomass production and a seasonally changing oxygen minimum zone. Nitrate:phosphate ratios in fresh upwelling water are low due to denitrification in the near‐seafloor oxygen minimum zone and phosphate efflux from sediments. This makes the region a candidate for substantial dinitrogen fixation, for which evidence is scarce. Nutrient and oxygen data, N isotope data of nitrate, nitrogen isotope ratios of particulate matter, particulate organic carbon content, and suspended matter concentrations on a transect across the shelf and upper slope at 23°S illustrate N‐cycling processes and are the basis for estimating the contribution of N‐sources and N‐sinks to the reactive nitrogen pool. It appears that N‐removal due to denitrification exceeds N gain by N2 fixation and physical mixing processes by a factor of >6, although inorganic N:P ratios again increase as surface water is advected offshore. Nitrate and ammonium regeneration, nutrient assimilation with N:P < 16, shelf break mixing, atmospheric input, and N2 fixation all contribute to the restoration of inorganic N:P ratios back to Redfield conditions, but in seasonally changing proportions. The Benguela upwelling system thus is a nutrient source for the oceanic‐mixed layer where N‐sources and N‐sinks are not in balance and Redfield conditions can only re‐adjust by advection and mixing processes integrated over time

Geranylphenazinediol, an Acetylcholinesterase Inhibitor Produced by a Streptomyces Species

Geranylphenazinediol (1), a new phenazine natural product, was produced by the Streptomyces sp. strain LB 173, which was isolated from a marine sediment sample. The structure was established by analysis of NMR and MS data 1 inhibited the enzyme acetylcholinesterase in the low micromolar range and showed weak antibacterial activity. In order to get a more detailed picture of the activity profile of 1, its inhibitory potential was compared to that of related structures

Simultaneous quantification of fentanyl, sufentanil, cefazolin, doxapram and keto-doxapram in plasma using liquid chromatography–tandem mass spectrometry

A simple and specific UPLC–MS/MS method was developed and validated for simultaneous quantification of fentanyl, sufentanil, cefazolin, doxapram and its active metabolite keto-doxapram. The internal standard was fentanyl-d5 for all analytes. Chromatographic separation was achieved with a reversed-phase Acquity UPLC HSS T3 column with a run-time of only 5.0 min per injected sample. Gradient elution was performed with a mobile phase consisting of ammonium acetate or formic acid in Milli-Q ultrapure water or in methanol with a total flow rate of 0.4 mL min−1. A plasma volume of only 50 μL was required to achieve adequate accuracy and precision. Calibration curves of all five analytes were linear. All analytes were stable for at least 48 h in the autosampler. The method was validated according to US Food and Drug Administration guidelines. This method allows quantification of fentanyl, sufentanil, cefazolin, doxapram and keto-doxapram, which is useful for research as we

PhosPhAt: the Arabidopsis thaliana phosphorylation site database. An update

The PhosPhAt database of Arabidopsis phosphorylation sites was initially launched in August 2007. Since then, along with 10-fold increase in database entries, functionality of PhosPhAt (phosphat.mpimp-golm.mpg.de) has been considerably upgraded and re-designed. PhosPhAt is now more of a web application with the inclusion of advanced search functions allowing combinatorial searches by Boolean terms. The results output now includes interactive visualization of annotated fragmentation spectra and the ability to export spectra and peptide sequences as text files for use in other applications. We have also implemented dynamic links to other web resources thus augmenting PhosPhAt-specific information with external protein-related data. For experimental phosphorylation sites with information about dynamic behavior in response to external stimuli, we display simple time-resolved diagrams. We have included predictions for pT and pY sites and updated pS predictions. Access to prediction algorithm now allows ‘on-the-fly’ prediction of phosphorylation of any user-uploaded protein sequence. Protein Pfam domain structures are now mapped onto the protein sequence display next to experimental and predicted phosphorylation sites. Finally, we have implemented functional annotation of proteins using MAPMAN ontology. These new developments make the PhosPhAt resource a useful and powerful tool for the scientific community as a whole beyond the plant sciences