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Lecture 19 Spectrophotomet ry-III
16

Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Dec 20, 2015

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Page 1: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Lecture 19

Spectrophotometry-III

Page 2: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Light Source

Sample

Monochromator(filter,

wavelength selector)

Detector

Spectrometer

Data Processing

Page 3: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Single beam

Double beam

Page 4: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

A tungsten lamp

3000 K

Page 5: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Deuterium Lamp

hHHHEH '''202

Deuterium: higher intensity

UV sources

Page 6: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Limits to Beer’s Law

strayP

strayPT

0

1

Incident light

P0

Emergent light

P1

Stray light

Page 7: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.
Page 8: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Solvents for UV/vis Spectrophotometry

Solvent Lower Wavelength Limit

Water 180 nm

Ethanol 220 nm

Hexane and 200 nmCyclohexane

Acetone 320 nm

Page 9: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.
Page 10: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

AClT logBeer’s law Bugert, Lambert and Beer

A

concentration

Straight line!

Page 11: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Wavelength, nm

Aor

B

Other components of the mixture

Measure @this wavelength!

Page 12: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Limits to Beer’s Law

Polychromatic light (several wavelength)

Page 13: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Wavelength, nm

Aor

B

Other components of the mixture

Intensity is too low!

Page 14: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Wavelength, nm

Aor

B

Other components of the mixture

Measurement is difficult

Page 15: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

Wavelength, nm

Aor

C

Other components of the mixture

Measure @this wavelength!

B + reagent CCompound is chemically derivitized (transformed) prior to themeasurement of absorbance

Page 16: Lecture 19 Spectrophotometry- III. Light Source Sample Monochromator (filter, wavelength selector) Detector Spectrometer Data Processing.

C2H5N

S-

SC2H5Cu

S

S S

SN

C2H5

C2H5

NC2H5

C2H52 +

Cu2+

NH

NN

S

N

Hg

N

S

NN

HN

N

H

N N

SH

NHg2+

NO2

NO2

HNNH2

+O

NO2

NO2

HNN