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Samenvatting Instrumentele chemie 2de jaar BMLT B. Muyssen
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  • Course
  • Instrumentele Chemie
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  • Hogeschool Gent (HoGent)

Samenvatting Instrumentele chemie 2de jaar BMLT B. Muyssen

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  • February 6, 2023
  • 28
  • 2021/2022
  • Summary

Subjects

  • spectrochemie
  • massaspectometrie
  • elektrochemie
  • chromatografie
  • instrumentele chemie

Written for

  • Hogeschool Gent (HoGent)
  • Biomedische Laboratoriumtechnologie
  • Instrumentele Chemie
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DEEL I: SPECTROCHEMIE
Inhoudsopgave
1) UV/VIS spectrofotometrie.............................................................................................. 3
1. Algemeen.......................................................................................................................................... 3

2. Principe spectrofotometer................................................................................................................. 3
Wet van Lambert-Beer....................................................................................................................... 4

3. Onderdelen toestel............................................................................................................................ 4
4. Instrumentontwerp............................................................................................................................ 4

5. Meetresultaten en datamanipulatie.................................................................................................. 5
6. Toepassing van UV-VIS...................................................................................................................... 5

7. Reflectiefotometrie........................................................................................................................... 5

2) Nefelometrie & turbidimetrie........................................................................................7

1. Principe............................................................................................................................................. 7
2. Onderdelen....................................................................................................................................... 7

3. Nefelometrie wetten.......................................................................................................................... 7
4. Turbidimetrie..................................................................................................................................... 7

5. Toepassingen.................................................................................................................................... 7

3) Infrarood...................................................................................................................... 8

1. Principe............................................................................................................................................. 8
2. Inleiding............................................................................................................................................ 8

3. Apparatuur........................................................................................................................................ 8
4. Toepassingen.................................................................................................................................... 8

4) Vlamfotometrie = Atomaire Emissie Spectometrie (AES)................................................9
1. Principe............................................................................................................................................. 9

2. Processen in een vlam...................................................................................................................... 9
3. Onderdelen....................................................................................................................................... 9

4. Storingen........................................................................................................................................... 9
5. ICP-AES........................................................................................................................................... 10

5) Atomaire Absorptie Spectometrie (AAS).......................................................................11
1. Principe........................................................................................................................................... 11

2. Onderdelen..................................................................................................................................... 11
3. Storingen......................................................................................................................................... 11

4. Toepassing...................................................................................................................................... 11
5. Verschillen met AES........................................................................................................................ 12

6) Fluorimetrie............................................................................................................... 12

, 1. Inleiding.......................................................................................................................................... 12

2. Theorie van fluorescentie................................................................................................................ 12
3. Onderdelen..................................................................................................................................... 12

6. Verschillen met AES........................................................................................................................ 13

7) Nucleaire magnetische resonantie (NMR).....................................................................13

1. Inleiding.......................................................................................................................................... 13
2. Onderdelen..................................................................................................................................... 13

1) Principe..................................................................................................................... 14

2) Onderdelen apparatuur............................................................................................... 14

1. Inbrengen monster.......................................................................................................................... 14
2. Ionisatiebron................................................................................................................................... 14

3. Massascheiding – analysator........................................................................................................... 14
4. Detectie.......................................................................................................................................... 14

1) Inleiding..................................................................................................................... 19

2) Conductometrie.......................................................................................................... 19

3) Potentiometrie........................................................................................................... 19
1. Inleiding.......................................................................................................................................... 19
A) Galvanische cellen....................................................................................................................... 19
B) Elektrodepotentiaal...................................................................................................................... 19
C) Standaard elektrodepotentiaal = SEP = E0..................................................................................19
D) Elektromotorische kracht (EMK) van galvanische cellen..............................................................19
E) Vloeistofjunctiepotentiaal............................................................................................................. 20
2. Potentiometrie................................................................................................................................ 20
A) Inleiding....................................................................................................................................... 20
B) Instellen ionensterkte.................................................................................................................. 20
C) Selectiviteit.................................................................................................................................. 20
D) Meetlimieten................................................................................................................................ 20
E) Responstijd.................................................................................................................................. 20
3. Referentie-elektrode....................................................................................................................... 20
A. Verzadigde calomelelektrode (Hg2Cl2)........................................................................................20
B. Zilver-zilverchloride elektrode (AgCl)........................................................................................... 20
C. Kwik-kwiksulfaat elektrode (Hg2SO4)...........................................................................................20
4. Ion selectieve elektrode (ISE).......................................................................................................... 21
A. Glasmembraan selectief voor H+  pH meter................................................................................21
B. Glasmembraan selectief voor Na+...............................................................................................21
C. Vaste kristalmembraan voor Cl-................................................................................................... 21
D. Vloeistofmembraan voor K+........................................................................................................ 21
E. Glasgevoelige elektrode voor CO2............................................................................................... 21
F. Enzymelektrode............................................................................................................................ 21

5. Analytische potentiometrische methoden.......................................................................................21

4) Ampèro, voltam- en coulometrie..................................................................................22

A. Ampèrometrie................................................................................................................................. 22

, B. Voltammetrie.................................................................................................................................. 22

C. Coulometrie.................................................................................................................................... 22

5) Biosensoren............................................................................................................... 22

1) Inleiding..................................................................................................................... 26

2) Uitwisselingsprincipes................................................................................................26

1. Adsorptiechromatografie................................................................................................................. 26
2. Gebonden fase................................................................................................................................ 26

3. Ionenuitwisselings........................................................................................................................... 26
4. Verdelings....................................................................................................................................... 26

5. Affiniteits......................................................................................................................................... 26
6. Gelfiltratie....................................................................................................................................... 26

3) Staalvoorbereiding..................................................................................................... 27

4) Standaardisatie.......................................................................................................... 27

5) Dunne laag chromatografie (TLC)................................................................................27

6) Conventionele LSC...................................................................................................... 27

7) HPLC.......................................................................................................................... 27
Vergelijking conventionele LSC en HPLC............................................................................................. 27

8) Gaschromatografie..................................................................................................... 28

9) Technologische ontwikkelingen...................................................................................28



1) UV/VIS spectrofotometrie
1. Algemeen
- UV licht  200-400 nm
- Zichtbaar licht  400-800 nm
- E=h.v
o v = frequentie
- Blauw  meest energie
- Rood  minste energie

2. Principe spectrofotometer


M + h . v  M*  M + warmte
- Molecule waar je elektromagnetische straling aan toevoegt en dan krijg je een geëxciteerde
molecule die dan terugvalt in de grondtoestand met vrijkomen van een beetje warmte. Het licht
(h.v) is verdwenen/geabsorbeerd en dat gaan we meten met spectrofotometer.
- * = geëxciteerd
- h . v = elektromagnetische straling

,Wet van Lambert-Beer
- A=ε λ . C . b
I0
- A=log
I

3. Onderdelen toestel
1) Lichtbron = stralingsbron
a. VIS  wolfraam(tungsten)-halogeen
b. UV  H2 of deuterium
c. UV/VIS  xenon
2) Monochromator = golflengteselector
a. Filters
i. Glasfilter = absorptiefilter
ii. Interferentiefilter (in fase:
versterken ; niet in fase: uitdoven)
b. Monochromators
i. Rooster
3) Cuvet = staaltje
a. Glas, plastic, kwarts
4) Detector
a. Fotomultiplicatorbuis (gevoelig maar plaatsinnemend)
b. Siliciumfotodiode (minder gevoelig)
i. Fotodiode array
5) Recorder = data output




4. Instrumentontwerp
1) Visuele/vergelijkende colorimetrie
a. Set testbuizen van identieke afmetingen  vergelijken met standaardoplossingen
b. Lovibond schijf comparator  schijf gekleurde glazen filters met kleurschaal geijkt op
conc.
c. Dusbosq colorimeter  1 standaardoplossing door wijziging weglengte (diepte)
2) Foto-elektrische colorimeter  absorptie uit VIS met filters en ronde glazen cuvet (brede A
pieken)
3) Spectrofotometer  toestel met roostermonochromator om elke λ te kunnen selecteren
a. Single beam = enkelstraal  1 ding tegelijk (blanco apart)
i. +  Hoge lichtopbrengst, lage ruis, weinig strooilicht, goedkoop
ii. -  niet voor kin exp en kwalitatieve studies met continue meting A, tijdrovend,
drift
iii. LB  M  C  D
b. Double beam = dubbelstraal  veranderingen in oplossing (blanco tegelijk)
i. +  korte kalibratie, geringe drift
ii. LB (dubbele lichtstraal)  M  CHOPPER  2 C  1D (FMB)
c. Dual beam = gesplitst
i. +  geen chopper (betrouwbaarheid), fotodiode ipv FMV vereist geen hoog V
ii. -  fotodiode minder gevoelig (A>3  verdunnen of kleinere d)
iii. LB (dubbele lichtstraal)  M  BEAM SPLITTER  2 C  2 D (fotodiode)

, d. Stable beam  enkelstraal met λ scan
4) Fototrode  lijkt op elektrode, in oplossing, storingen geëlimineerd, titraties

5. Meetresultaten en datamanipulatie
1) Vaste golflengte analyse
a. Eindpunttesten  A, %T, conc
b. Kinetische testen  enzymactiviteit
c. A verhouding en -verschil  zuiverheid (A260/280  DNA)
d. 3 punten netto-A (Allen basislijnmethode)
e. Multipele golflengte analyse
f. Statistiek en testprogramma’s
2) Scanning analyse  dubbelstraal, enkelstraal met referentiescan
a. Herhaalde scans
b. Automatische pieklocatie (identificatie absorberende comp in staal
c. 3 punt netto 1 slechte basislijn

6. Toepassing van UV-VIS
1) Kwalitatieve analyse
a. Oplosmiddel  beter apolair
b. Spectrale bandbreedte (en spleetopening) beter klein
c. Strooilicht
2) Kwantitatieve analyse
a. Reagentia  A verhogen
b. Selectie golflengte  meten bij λ max (absorptiepiek)
c. Verband A en c  mol abs ε uit ijklijn beter dan uit 1 punt of literatuur
d. Fotometrische precisie
i. Fouten kleinst tussen 10-70 %T en dus 0,155-1 A
3) Fotometrische titraties
4) Problemen bij fotometrische bepalingen
a. Correcties
i. Bij 1 golflengte de A optellen en aftrekken
 A gemeten= A bepaling + A reagens + (A troebeling + A kleur andere
stoffen)
 A bepaling= A gemeten – A reagens – (A troebeling + A kleur andere
stoffen)
ii. A reagens  reagens blanco 1x
iii. A troebeling + A kleur andere stoffen  reagens monsterblanco elke keer
b. Troebelheid
c. Afwijkende kleur
d. Bichromatisch meten ipv blanco  bij 2 golflengtes (best waarbij interferentie gelijk is)

7. Reflectiefotometrie
- Spectrofotometer = filterfotometer
 Heldere oplossing
 Transmissie/absorbantie met monochromatisch licht door cuvet meten
 LB voor conc bepaling
- Reflectiefotometer = droge chemie analysers
 Vaste stoffen
 Hoeveelheid licht teruggekaatst bij bepaalde golflengte
 Teststrookjes
 Kubelka-Munk voor correlatie conc en refl

,

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