Optimization of sample recovery from the nitrocellulose support used in plasma desorption mass spectrometry and its use for multiple analyses of insulin
| Autor: | Sonja Jespersen, Gert Talbo, Peter Roepstorff |
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| Rok vydání: | 1993 |
| Předmět: |
Molecular Sequence Data
Analytical chemistry Mass spectrometry Biochemistry High-performance liquid chromatography Mass Spectrometry Iodine Radioisotopes chemistry.chemical_compound Adsorption Desorption Mass transfer Animals Humans Insulin Amino Acid Sequence Cysteine Chromatography High Pressure Liquid Spectroscopy Chromatography Extraction (chemistry) Difluoride Collodion Membranes Artificial Dithiothreitol chemistry Molecular Medicine Cattle Polyvinyls Oxidation-Reduction Nitrocellulose |
| Zdroj: | Biological Mass Spectrometry. 22:77-83 |
| ISSN: | 1096-9888 1052-9306 |
| DOI: | 10.1002/bms.1200220110 |
| Popis: | The parameters for recovery of sample from the nitrocellulose support used in plasma desorption are optimized. The losses in washing, in situ reactions, and extraction procedures are quantitatively evaluated. At least 80% of the sample can be effectively extracted or transferred to a polyvinyl difluoride membrane with 2-propanol-water mixture in the range 1:1-2:3 v/v. Quantitative losses of insulin during washing procedures vary from 0 to 50% depending on the washing procedure used. The losses in in situ reactions are negligible. Optimization of the procedures allows several successive procedures to be carried out after adsorption of 1 nmol of insulin on the nitrocellulose support. These include in situ reaction, extraction of A and B chains followed by S-alkylation, chain separation by high-performance liquid chromatography, mass spectrometric analysis of the separated chains, and finally automatic sequence after transfer to the sequenator. |
| Databáze: | OpenAIRE |
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