ABSORPTION SPECTRA AND BEER

LAMBERTS LAW
G1 (GROUP 3)
Bolotaolo, Trisha Mae V.
Buluran, Cherri Marie
Boo, Dahee
Bulaong, Gem Bernadette
Caba, Kaye
Objectives:
To demonstrate the principles of
spectrophotometry
To plot the absorption spectra of
Bromphenol Blue and Methyl orange
To identify the point of maximal
absorption of BPB and MO.
Ultraviolet-Visible Absorption Spectrophotometry
 Wavelength

= c/v

Plancks formula: E= hv
Rayleigh Scattering and Absorption
Rayleigh scattering- photon is
diffracted/scattered with unchanged
frequency

Absorption- photon transfers its energy to the

molecule
SPECTROPHOTOMETRY
Spectrophotometer
- measures amount of light
transmitted by a sample.
 Principle
Light absorbed= concentration
How does a spectrophotometer work?
 Components of a
spectrophotometer
LIGHT SOURCE
 LIGHT SOURCE
provides polychromatic light and must generate
sufficient radiant energy to measure the
analyte of interest.

Examples: Tungsten bulb, Deuterium lamp and

Hydrogen lamp
 ENTRANCE SLIT

- minimizes unwanted or stray light and

prevents the entrance of scattered light into
the monochromator system
MONOCHROMATOR
 MONOCHROMATOR
Isolates specific or individual wavelength of
light

Kinds:
1. Prism- refraction
2. Diffraction gratings- diffraction
3. Filter- constructive interference of waves
 EXIT SLIT

Controls the width of light beam

Allows only a narrow fraction of the spectrum to
reach the sample cuvette
CUVETTE (SAMPLE CELL)
 CUVETTE (SAMPLE CELL)
absorption cell/analytical cell/ sample cell
Holds the solution whose concentration is to
be measured
 CUVETTE (SAMPLE CELL)
Kinds:
Soft Glass Cuvette: for acidic solution; UV
Borosilicate Cuvette: for strong alkaline
solution
Quartz or Plastic Cuvette: for UV and VIS
Disposable: polymethacrylate and polstyerene
 DETECTOR

- detects and converts transmitted light into

photoelectric energy
 Examples:
Photomultiplier tube- most common
Photodiodes- modern spectrophotometers
READ-OUT DEVICE
 Read-out Device
- displays output of the detection system

Examples: Galvanometer, Ammeter, Recorder,

Monitor