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A folyadékkromatográfia -
tömegspektrometria (LC-MS)
elve és lehetőségei
Folyadékkromatográfia - tömegspektrometria (LC-MS)
• analitikai kémiai technika, amely a folyadékkromatográfiás (LC) fizikai
elválasztási képességeit és a tömegspektrometria (MS) tömegelemzési
képességeit ötvözi
• az 1980-as években indult
• sok vegyület mérhető egyetlen analitikai futtatással
• nagyon érzékeny és szelektív
• hasznos számos alkalmazásban
Komponensek
Folyadék
kromatográfia
Ion Forrás
Tömeg
elemző
Detektor
Folyadékkromatográfia
a) oldószerbevezető szűrő, (b) szivattyú, (c) inline oldószerszűrő, (d) befecskendező
szelep, (e) előoszlopos szűrő, (f) oszlop, (g) detektor, (h) felvevő, (i)
hátnyomásszabályozó j) hulladéktartály
Tömegspektrometria
Ion forrás
tömeg
elemző
Detektor
PHOTO Dario
Tömegspektrometria
Minta
injektálás Egység
Ionizációs
forrás
tömeg
analizátor
Detektorrendszer
Adatrendszer
Ion létrehozás
Adatok elemzése
Ionok tömeg szerinti osztályozása (m / z)
Érzékelés
• szilárd
• folyékony
• gőz
Ionizáció és interfész
Elektrospray ionizáció (ESI)
Légköri nyomás kémiai ionizáció (APCI).
Légköri nyomás fotionizáció(APPI)
Thermospray ionizáció(TSI)
Részecske sugár ionizáció(PBI)
Ionizáció és interfész
Elektrospray
ionizáció (ESI)
Ionization and Interface
Légköri nyomás
fotionizáció
Légköri nyomás kémiai
ionizáció (APCI)
Ionization and Interface
Thermospray
Ionizáció (TSI)
Ionization and Interface
Részecske sugár ionizáció(PBI)
Tömegelemzők
• A tömegspektrométer egy
összetevője
• az ionizált tömegeket elválasztja
a töltés / tömeg arányok alapján
• továbbítja őket az érzékelőhöz,
ahol azokat észlelik, majd később
digitális kimenetekké alakítják át.
Quadrupole
Analizátor
A
repüléselemző
ideje
Ion csapda
Analizátor
Hybrid
Analizátor
Tömeg analizátorok
Quadrupole analizátor
A repüléselemző ideje
Ion Trap Analyzer
Tömeg analizátorok
• Az ionok - az érzékelő által elektromosan érzékeltek (tömeg / töltési arányuk szerint vannak
elválasztva)
• Az érzékelő kiválasztása a következőkön alapul:
• a szükséges érzékelési érzékenység és a sebesség
Detektorok
Legkevésbé
érzékeny
Faraday kupa
detektor
1000x nagyobb
érzékenység a Faraday
csészéhez képest
Fénysokszorozó
detektor
Legérzékenyebb.
Nagy felbontású
Fényképészeti
detektor
Detektorok
Faraday kupa detektor
Detektorok
Fényképészeti detektor
Detektorok
Fénysokszorozó detektor
Adatfelvétel
Teljes tömegspektrum - SCAN technika
Kiválasztott ionfigyelés- SIM technika
Kiválasztott ionfigyelés- SIM technika
Alkalmazások Gyógyszerészeti
elemzés
Biohasznosulási
vizsgálatok
Gyógyszer
bomlástermék-
elemzés
A potenciális
gyógyszerek
szűrése és
jellemzése
Gyógyszer-
metabolizmus
vizsgálatok,
farmakokinetika
A gyógyszer
targetek
azonosítása
Biomolekula
jellemzése
Fehérjék és
peptidek
oligonukleotidok
Törvényszéki
elemzés
Környezeti
elemzés
Peszticidek
élelmiszereken
Talaj- és talajvíz
szennyezés

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Editor's Notes

  1.   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).
  2.   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).