Isoprostanes Analysis Service


Isoprostanes are a newly  discovered class of compounds formed in vivo as a consequence of non-enzymatic  oxidation of polyunsaturated fatty acids. The isoprostanes produced by free  radical-catalyzed peroxidation of arachidonic acid, so called F2-isoP, were the  first to be introduced by Mihelich. The discovery of isoprostanes has important  evidences for medicine. Their occurrence has been established as the most  predictable marker of oxidative stress, while the measurement of their  concentration in biological samples is the most recommended approach to the  assessment of redox status in vivo, providing a crucial tool to explore the  role of oxidative processes in the pathogenesis of human disease. The elevated  levels of F2-isoP  have been observed in many conditions associated with increased abundance of  reactive oxygen species such as asthma, neurodegenerative diseases (Huntington,  Alzheimer or multiple sclerosis), alcoholic and non-alcoholic liver diseases,  pulmonary disease or diabetes.

Biological function for isoprostanes  Figure  1 Biological function for isoprostanes.

Except  for F2-isoP,  there exist other types of isoprostanes found in vivo, including F3-isoP formed from  eicosapentaenoic acid (EPA) and the most structurally diverse  products of docosahexaenoic acid (DHA) non-enzymatic oxidation. The EPA- and  DHA-derived isoprostanes seem to have multiple biological roles, so F2-isoP remains the primary  indicators of oxidative stress.

The  isoprostanes are usually measured in plasma or urine by well-established gas  chromatography–mass spectrometry (GC-MS) and enzyme-linked immunosorbent assays  (ELISA). Even if the GC-MS approach is sensitive, the procedure is usually  tedious and time-consuming due to the fact that a derivatization step is  required prior to separation. Typically, isoprostanes are converted to  pentafluorobenzyl esters by treatment with pentafluorobenzyl bromide. The main  disadvantage of commercially available ELISA is that a cross-linking reaction  can be observed and each assay kit can only measure just one isomer of F2-isoprostanes.  However, when sensitivity and selectivity issues in the analysis of  biomolecules arise, among various chromatographic methods, liquid  chromatography-tandem mass spectrometry (LC-MS/MS) is optimal to overcome such  problems.

The  advantages of LC-MS/MS method include relatively higher sensitivity and  selectivity and short run times, especially in the case of such samples as  e.g., plasma that contain a large amount of coeluting interferences. LC-MS/MS  method has been proven to be very useful for simultaneous measurement of  different isomers of F2-isoprostanes.  What’s more, to achieve accurate quantitation at extremely low concentrations,  there is the possibility of using nonradioactive isotope-labeled standards to  compensate for the loss of analyte during sample preparation, which has been  the most important step for eliminating the matrix effect for analysis of  biomarkers. With experienced scientists at Creative Proteomics, we have  developed a reliable and reproducible method using highly sensitive LC-MS/MS method  for the rapid identification and quantification of diverse isoprostanes in  different sample types, which can satisfy the needs of academic and industrial  study in your lab.

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Isoprostanes Quantified in This Service

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Staffed by experienced biological scientists,  Creative Proteomics can provide a wide range of services ranging from the  sample preparation to the metabolite extraction, characterization,  identification and quantification. We are willing to provide our customer the  outstanding isoprostanes analysis service with accurate and reliable results,  quick turnaround, saving you time and resources! Please feel free to  contact us to discuss the wide range of services we can perform.


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