Journal of chromatography. B, Analytical technologies in the biomedical and life sciences最新文献

{"title":"HPLC analysis of human erythrocytic glutathione forms using OPA and N-acetyl-cysteine ethyl ester: evidence for nitrite-induced GSH oxidation to GSSG.","authors":"Jan Thomas Michaelsen,&nbsp;Sabine Dehnert,&nbsp;Daniela Giustarini,&nbsp;Bibiana Beckmann,&nbsp;Dimitrios Tsikas","doi":"10.1016/j.jchromb.2009.06.043","DOIUrl":"https://doi.org/10.1016/j.jchromb.2009.06.043","url":null,"abstract":"<p><p>Glutathione exists in biological samples in the reduced form (GSH), as its disulfide (GSSG) and as a mixed disulfide (GSSR) with thiols (RSH). GSH is the most abundant low-molecular-mass thiol and plays important roles as a cofactor and as a main constituent of the intracellular redox status. Due to its own sulfhydryl (SH) group, GSH reacts readily with o-phthaldialdehyde (OPA) to form a highly stable and fluorescent isoindole derivative (GSH-OPA), which allows for sensitive and specific quantitative determination of GSH in biological systems by HPLC with fluorescence (FL) detection. In the present article we report on the utility of the novel, strongly disulfide bond-reducing thiol N-acetyl-cysteine ethyl ester (NACET) for the specific quantitative analysis of GSH and GSSG in the cytosol of red blood cells (RBC) as GSH-OPA derivative with FL (excitation/emission 338/458nm) or UV absorbance (338nm) detection. Unlike in aqueous solution, the derivatization of GSH in RBC cytosol yielded two closely related derivatives in the absence of NACET and only the GSH-OPA derivative in the presence of NACET. The HPLC method was optimized and validated for human RBC and applied to measure GSH and GSSG in RBC of healthy subjects. Basal GSH and GSSG concentrations were determined to be 2340+/-350microM and 11.4+/-3.2microM, respectively, in RBC of 12 healthy young volunteers (aged 23-38 years). The method was also applied to study the effects of nitrite on the glutathione status in intact and lysed human RBC. Nitrite at mM-concentrations caused instantaneous and considerable GSSG formation in lysed but much less pronounced in intact RBC. GSH externally added to lysed RBC inhibited nitrite-induced methemoglobin formation. Our findings suggest that nitric oxide/nitrite-related consumption rate of GSH, and presumably that of NADH and NADPH, could be of the order of 600micromol/day in RBC of healthy subjects.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"3405-17"},"PeriodicalIF":3.0,"publicationDate":"2009-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jchromb.2009.06.043","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40021565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Proceedings of the 33rd Annual Meeting of the Japanese Society for Biomedical Mass Spectroscopy. September 25-26, 2008. Tokyo, Japan.","authors":"","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"2599-661"},"PeriodicalIF":3.0,"publicationDate":"2009-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40042125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Methodologies for the transfer of analytical methods: a review.","authors":"E Rozet,&nbsp;W Dewé,&nbsp;E Ziemons,&nbsp;A Bouklouze,&nbsp;B Boulanger,&nbsp;Ph Hubert","doi":"10.1016/j.jchromb.2008.12.049","DOIUrl":"https://doi.org/10.1016/j.jchromb.2008.12.049","url":null,"abstract":"<p><p>The transfer of a method from a laboratory to a production site is an important step in the development cycle of new pharmaceutical products. Method transfers are increasingly implemented due to the economical pressure coming from the rationalization of production sites, analytical subcontracting and fusion of pharmaceutical groups. However, no official guidance regarding study design, data analysis, or decision procedures is present neither in FDA documents nor in ICH documents for method transfers. The experiments performed in such a transfer and the methodology used to accept or reject it should be fitted for purpose. In order to provide to analysts a global view of the problematic of analytical method transfer, this paper reviews the documentation available in the scientific literature about the design of transfer studies and the required sample size. Special focus is also made on the statistical methodologies available for decision making with particular emphasis on risk management. Examples of transfer of pharmaceutical, bio-pharmaceutical and biological methods published in the literature are reviewed in order to illustrate the various possibilities among the strategies for methods transfer.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"2214-23"},"PeriodicalIF":3.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jchromb.2008.12.049","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39992202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fatal intoxications by acenocoumarol, phenprocoumon and warfarin: method validation in blood using the total error approach.","authors":"Raphaël Denooz,&nbsp;Zoénabo Douamba,&nbsp;Corinne Charlier","doi":"10.1016/j.jchromb.2008.11.049","DOIUrl":"https://doi.org/10.1016/j.jchromb.2008.11.049","url":null,"abstract":"<p><p>A simple high-performance liquid chromatography (HPLC) method with ultraviolet (UV) detection has been developed and validated for simultaneous identification and quantification of three antivitamin K drugs (acenocoumarol, warfarin and phenprocoumon) in whole blood. The aim of this development was to propose an analytical technique adapted to the situations of forensic toxicology, i.e. intoxication with massive anticoagulant doses, when the usual coagulation tests could not be used. The blood sample, after spiked with prazepam as an internal standard (IS), was submitted to a liquid-liquid extraction (LLE) prior to HPLC analysis. A chromatographic separation was achieved on a C8 Symmetry column with a mobile phase consisting of an acetonitrile and phosphate buffer (pH 3.8) mixture in a gradient mode. Detection was carried out at a wavelength between 200 and 400 nm. This method has been validated with the concept of total error as decision criterion. Trueness ranged from 99.1% to 105.0% and precision was good with RSD between 1.3% and 6.7%. Consequently, this rapid and simple chromatographic technique is well adapted to focus intoxications with most important coumarinic drugs available on pharmaceutical market and is now routinely used in our laboratory for forensic \"general unknown\" screening.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"2344-8"},"PeriodicalIF":3.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jchromb.2008.11.049","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39992203","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}