Samenvatting Scheidingstechnieken

INSTRUMENTELE ANALYSE 2:
SCHEIDINGSTECHNIEKEN
Lector. Opsteyn Ine




2023 - 2024

,INHOUD

1. Chromatografische technieken : inleiding .......................................................................................................... 3

1.1 Indeling van de chromatografische technieken ............................................................................................ 3

2. Gaschromatografie (GC) ...................................................................................................................................... 4

2.1 Verloop van een GC scheiding....................................................................................................................... 4

2.2 GC-scheidingsprincipe ................................................................................................................................... 4

2.3 Instrumentatie .............................................................................................................................................. 5

2.3.1 Dragergas ............................................................................................................................................... 5

2.3.2 Injectie .................................................................................................................................................... 5

2.3.3 Capillaire kolom...................................................................................................................................... 6

2.3.4 Oven ....................................................................................................................................................... 6

2.3.5 Detector – FID ........................................................................................................................................ 8

2.3.6 Registratie detectorsignaal .................................................................................................................... 9

Oefeningen ........................................................................................................................................................ 11

3. Vloeistof chromatografie (LC) ........................................................................................................................... 12

3.1 Algemeen .................................................................................................................................................... 12

3.2 Scheidingsprincipe ...................................................................................................................................... 12

3.2.1 Mobiele fase ......................................................................................................................................... 12

3.2.2 Stationaire fase .................................................................................................................................... 13

3.2.3 Injectie .................................................................................................................................................. 15

3.2.4 Pomp .................................................................................................................................................... 15

3.2.5 Detector ............................................................................................................................................... 16

4. Kwantifacatiemethoden .................................................................................................................................... 18

4.1 Interne standaard methode ........................................................................................................................ 18

4.1.1 Eenpunts kalibratie .............................................................................................................................. 18

4.1.2 Ijklijnmethode ...................................................................................................................................... 19

4.2 Standaardadditiemethode .......................................................................................................................... 19

4.2.1 eenpunts kalibratie .............................................................................................................................. 19

4.2.2 ijklijnmethode ...................................................................................................................................... 19


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, 5. Kwaliteit van scheidingen.................................................................................................................................. 21

5.1 het chromatogram ...................................................................................................................................... 21

5.2 Kwaliteit van scheidingen ............................................................................................................................ 21

5.2.1 Schotelgetal .......................................................................................................................................... 21

5.2.2 Resolutie............................................................................................................................................... 22

5.2.3 Assymetriefactor T (tailing factor) ....................................................................................................... 24

Oefeningen ........................................................................................................................................................ 25

6. hoge resolutie Elektroforese ............................................................................................................................. 27

6.1 Elektroforese als scheidingstechniek .......................................................................................................... 27

6.2 Gelelektroforese.......................................................................................................................................... 27

6.2.1 Scheidingsfactoren ............................................................................................................................... 27

6.2.2 Zuurtegraad van de oplossing .............................................................................................................. 27

6.2.3 Diffusie ................................................................................................................................................. 28

6.3 (Iso)-elektrofocusing (IEF) ........................................................................................................................... 28

6.3.1 Basisprincipe ........................................................................................................................................ 28

6.3.2 pH-gradiënt .......................................................................................................................................... 29

6.3.3 Apparatuur ........................................................................................................................................... 29

6.3.4 Dunlaag elektrofocusing ...................................................................................................................... 29

6.4 Gelelektroforese VS Elektrofocusering ....................................................................................................... 30




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