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HPLC- High-Performance Liquid Chromatography HPLC-High-Performance Liquid Chromatography

The High-Performance Liquid Chromatography Technique High-Performance Liquid Chromatography Technique

The analysis of peptides and proteins has been revolutionized by the introduction of HPLC. En analyse af peptider og proteiner er blevet revolutioneret af indførelsen af HPLC. HPLC allows for the rapid and sensitive analysis of peptide and protein structure. HPLC giver mulighed for hurtig og følsom analyse af peptid og protein struktur. It has simplified our analysis in understanding of biological processes and in the development of peptide and protein based pharmaceuticals. Det har forenklet vores analyse i forståelse af biologiske processer og i udviklingen af peptid og protein-baserede lægemidler.

But HPLC doesn't stop there, it's applications in peptide and protein purification continues to expand at an extremely rapid rate. Men HPLC stopper ikke her, det er ansøgninger i peptid og protein rensning fortsætter med at ekspandere i en meget hurtig sats. For example solid-phase peptide synthesis and recombinant DNA techniques have allowed the production of large quantities of peptides and proteins that need to be highly purified. For eksempel fast-fase peptid syntese og rekombinant DNA-teknik har tilladt produktion af store mængder af peptider og proteiner, som det er nødvendigt at være meget renset. Also another very important application for HPLC in the post-genomic age is that its techniques are used extensively in the isolation and characterization of novel proteins. Også et andet meget vigtigt ansøgning om HPLC i post-genomiske alder er, at dens teknikker er udbredt i isolation og karakterisering af nye proteiner.

HPLC's most significant feature would be its excellent resolution that is easily achieved under a wide range of conditions for very closely related molecules, as well as structurally quite distinct molecules. HPLC's vigtigste funktion ville være sin fremragende beslutningsforslag, som er let opnået under en bred vifte af betingelserne for meget nært beslægtede molekyler, såvel som strukturelt er meget forskellige molekyler. This feature is from the fact that all interactive modes of chromatography are based on recognition forces that can be subtly manipulated through changes in the elution conditions that are specific for the particular mode of chromatography. Denne funktion er fra det faktum, at alle interaktive former for kromatografi er baseret på anerkendelse kræfter, der kan raffineret manipuleres gennem ændringer i det eluerede betingelser, som er specifikke for den særlige form for kromatografi. Peptides and proteins interact with the chromatographic surface in an orientation specific manner, in which their retention time is determined by the molecular composition of specific contact regions. Peptider og proteiner interagerer med de chromatografiske overflade i en orientering særlige måde, hvorpå deres retentionstid bestemmes af den molekylære sammensætning af specifikke kontakt regioner. For larger polypeptides and proteins that adopt a significant degree of secondary and tertiary structure, the chromatographic contact region comprises a small proportion of the total molecular surface. For større polypeptider og proteiner, som indfører en betydelig grad af sekundære og tertiære struktur, chromatografisk kontakt region omfatter en lille del af den samlede molekylære overflade. All biological processes depend on specific interactions between molecules and affinity chromatography exploits these specific interactions to allow the purification of a biomolecule on the basis of its biological function or individual chemical structure. Alle biologiske processer er afhængige af specifikke interaktioner mellem molekyler og affinitet kromatografi udnytter disse specifikke interaktioner at tillade rensning af en biomolecule på grundlag af dets biologiske funktion eller individuelle kemiske struktur.

Retention and Bandwidth Relationships of HPLC Lagring og Båndbredde Forhold af HPLC

Retention time or tr in HPLC is the time taken for a solute to pass through a chromatographic column . Retentionstid eller tr i HPLC er den tid, det tager for et opløst til at passere gennem en kromatografikolonnen. This retention time is measured as the time taken by the solute, following injection, to emerge from the column and to be detected. Denne retentionstid er målt som den tid, som opløst, efter injektion, at komme ud af kolonnen og til at blive opdaget. In order to allow retention times to be compared to different columns or under different conditions, the retention time of a solute is normally compared with the retention time of a molecule which is not retained on the specific column of interest. For at muliggøre retentionstider skal sammenlignes med forskellige kolonner eller under forskellige betingelser, retentionstiden af et opløst stof er normalt sammenlignes med retentionstid på et molekyle, der ikke beholdes om den særlige kolonne af interesse. This allows the unitless capacity factor k' of a solute to be expressed in terms of the retention time tr, through the relationship Dette gør det muligt for unitless kapacitet faktor k 'af et opløst stof, der skal udtrykkes i retentionstiden tr, gennem forholdet

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