Determination of 4-hydroxy-2-nonenal in primary rat hepatocyte cultures by liquid chromatography with laser induced fluorescence detection

4‐Hydroxynonenal: A membrane lipid oxidation product of medicinal interest

“…A sensitive HPLC method with laser-induced fluorescence detection was developed for the determination of free HNE in cells. 57 Carbonyl compounds were fluorescently labeled by incubating rat hepatocytes with a tagging reagent, 4-(2-carbazoylpyrrolidin-1-yl)-7-nitro-2,1,3-benzoxadiazole. The hydrazone derivatives were extracted with a solid phase extraction cartridge and separated on a reversed-phase HPLC column.…”

Shotgun Lipidomics Analysis of 4-Hydroxyalkenal Species Directly from Lipid Extracts after One-Step in Situ Derivatization

“…4-HNE could be derivatized with 2,4-dinitrophenylhydrazine (DNPH) 13,15 or fluorescently labeled with 4-(2-carbazoylpyrrolidin-1-yl)-7-nitro-2,1,3-benzoxadiazole. 14 The derivatives were then separated by HPLC and detected by electrochemical or laser-induced fluorescence detection to increase the sensitivity of the methodologies. Those methods did not adequately provide structural details regarding the 4-hydroxyalkenals.…”

“…Varieties of methodologies have recently been developed and used to identify and quantitate 4-HNE alone or along with other aldehydes present in biological samples, including gas chromatography/mass spectrometry (GC/MS), − high-performance liquid chromatography (HPLC), − HPLC combined with GC/MS, electrospray ionization (ESI)-MS after direct infusion, and LC–MS/MS. − In the methods using GC/MS, 4-HNE and other aldehydes were converted into their O -pentafluorobenzyloxime derivatives and then detected by selected ion monitoring (SIM) , or stable isotope dilution MS. , These approaches involved laborious multistep chemical modification. 4-HNE could be derivatized with 2,4-dinitrophenylhydrazine (DNPH) , or fluorescently labeled with 4-(2-carbazoylpyrrolidin-1-yl)-7-nitro-2,1,3-benzoxadiazole . The derivatives were then separated by HPLC and detected by electrochemical or laser-induced fluorescence detection to increase the sensitivity of the methodologies.…”

The fabrication and cell culture of three-dimensional rolled scaffolds with complex micro-architectures

“…Hepatocytes from SD rats of weight 160-180 g (First Affiliated Hospital of Medical Collage of Xi'an Jiaotong University, China) were isolated using a two-step collagenase perfusion procedure [33]. The density of the cell suspension was 2 × 10 6 cells ml −1 ; trypan-blue staining showed that the cell viability was over 80%.…”