Princip i izvedba tekućinske kromatografije-masene spektrometrije (LC-MS)
1. Princip i izvedba tekućinske
kromatografije-masene
spektrometrije (LC-MS)
2. Tekuća kromatografija - masena spektrometrija (LC-MS)
• Tehnika analitičke kemije koja kombinira fizičke mogućnosti razdvajanja
tekućinske kromatografije (LC) sa sposobnostima masene analize masene
spektrometrije (MS)
• uvedenih 1980-ih godina
• može se mjeriti u jednom analitičkom postupku ^ ^ vrlo visoko osjetljiva i
selektivna
• korisna u mnogim aplikacijama
6. Masena spektrometrija
Jedinica za
ubrizgavanje uzorka
Ionizacija
Izvor
Analizator mase
Sustav detektora
Sustav podataka
Ionska generacija
Analiza podataka
Razvrstavanje iona prema masi (m / z)
Otkrivanje
• Krutina
• tekući
• pare
12. Analizatori mase
• Komponenta masenog
spektrometra
• uzima i odvaja ionizirane mase
na temelju omjera naboj i masa
• ih izlazi u detektor gdje se
detektira i kasnije pretvara u
digitalni izlaz.
Quadrupole
analizator
Vrijeme
analizatora
leta
Ionska
zamka
analizator
Hybrid
analizator
14. Time of Flight Analyzer
Ion Trap Analyzer
Analizatori mase
15. • Iona - električki otkrio detektor (su odvojeni prema omjer njihovih masa/naboj)
• Izbor detektor temelji se na:
• potrebno otkrivanje osjetljivosti i brzini
• Toplinska i kemijska postojanost
Detectors
Najmanje
osjetljiv
Faraday kup
detektor
1000 x veću
osjetljivost u
odnosu na
Faradaya kup
Fotomultiplikacij
skom detektor
Najosjetljiviji.
Koristi se za
visoke
rezolucije
Fotografski
detektor
19. Snimanje podataka
Snimanje potpunog masenog spektra - SCAN tehnika
Praćenje odabranih iona - SIM tehnika
Praćenje odabranih iona - SIM tehnika
20. Prijave Farmaceutska
analiza
Studije
biodostupnosti
Analiza produkta
razgradnje lijeka
Probir i
karakterizacija
potencijalnih
lijekova
Istraživanja
metabolizma
lijekova,
farmakokinetika
Identificiranje
ciljeva lijeka
Karakterizacija
biomolekula
Proteini i
peptidi
Oligonukleotidi
Forenzička
analiza
Analiza
okoliša
Pesticidi na hrani
Onečišćenje tla i
podzemnih voda
Hinweis der Redaktion
ESI operates according to the following scheme:
1) the column effluent from the LC containing analytes is pumped through a capillary which is held at high potential (2-6 kV) and nebulized. The applied voltage can be either positive or negative, depending on the analytes; nebulization is usually assisted pneumatically (except in nanospray);
2) the droplets that detach from a tip of the capillary contain an excess of positive or negative charge as a result of the applied high voltage;
3) electrical field gradient attracts charged droplets towards the entrance of the mass spectrometer;
4) charged analyte molecules are generated from the small charged droplets either by the charged residue model or by the ion evaporation model. Ion formation from the droplets is promoted by a flow of drying gas (usually heated nitrogen);
5) the ions, solvent vapor and drying gas molecules are sampled through a capillary into a first pumping stage (0.08-0.75 Torr) where they are supersonically expanded;
6) the ions and some other neutral molecules are sampled via a skimmer into the second pumping stage (0.001-0.01 Torr) containing an ion focusing and transfer device (usually RF hexapole or octapole and a set of lenses);
7) ions enter mass analyzer region (< 10-5 Torr).
ESI operates according to the following scheme:
1) the column effluent from the LC containing analytes is pumped through a capillary which is held at high potential (2-6 kV) and nebulized. The applied voltage can be either positive or negative, depending on the analytes; nebulization is usually assisted pneumatically (except in nanospray);
2) the droplets that detach from a tip of the capillary contain an excess of positive or negative charge as a result of the applied high voltage;
3) electrical field gradient attracts charged droplets towards the entrance of the mass spectrometer;
4) charged analyte molecules are generated from the small charged droplets either by the charged residue model or by the ion evaporation model. Ion formation from the droplets is promoted by a flow of drying gas (usually heated nitrogen);
5) the ions, solvent vapor and drying gas molecules are sampled through a capillary into a first pumping stage (0.08-0.75 Torr) where they are supersonically expanded;
6) the ions and some other neutral molecules are sampled via a skimmer into the second pumping stage (0.001-0.01 Torr) containing an ion focusing and transfer device (usually RF hexapole or octapole and a set of lenses);
7) ions enter mass analyzer region (< 10-5 Torr).
