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Chem Lab 3 a Photo Metric Equipment

Chem Lab 3 a Photo Metric Equipment

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Published by biddyusmc
clinical chemistry
clinical chemistry

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Published by: biddyusmc on Jan 13, 2010
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07/21/2013

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MLAB 2401 - Clinical Chemistry Lab Manual
C
B 15
UNIT: Electromagnetic Radiation andPhotometric Equipment
3photo.wpd
Instrumentation I
Task
To review the theory of electromagnetic radiation and the principle and use of common laboratoryinstruments using electromagnetic radiation.
Objectives
Upon completion of this exercise, the student will be able to:1.State the wave and particle theory of electromagnetic radiation.2.Know what colors correspond to visible wavelengths.3.Identify the regions of the electromagnetic spectrum occupied by gamma, x-rays, UV, visible,IR, and microwaves indicating relative
wavelength
,
frequency 
, and
energy 
.4.Know basic principles of how instruments determine results.5.List and compare the principles/applications of different photometric equipment.
Principle
1.Properties of light and radiant energy.
Radiant energy 
behaves as if it has electric and magnetic fields and is found as
discretebundles
of energy (photons) traveling in
wave form
(particle and wave theories). The moreenergy contained, the more frequent the wave and therefore, the shorter the wavelength.
Wavelengths
are measured in nanometers (
nm
) or 10 meters.
-9
2.Interactions of light and matter. A.
Absorption
– (absorption spectroscopy – UV, visible, infra-red, atomic)When an atom, ion, or molecule absorbs a photon, the additional energy results in analteration of state (it becomes excited). Depending on the individual “species,” this maymean that a valence electron has been put into a higher energy level, or that thevibration or rotation of covalent bonds of the molecule have been changed.In order for a ray of radiation to be
absorbed 
it must:1.have the same frequency of the rotational or vibrational frequency in themolecules it strikes, and;2.be able to give up energy to the molecule it strikes.
 
UNIT: Electromagnetic Radiation and Photometric Equipment (continued)
B 16
C
MLAB 2401 - Clinical Chemistry Lab Manual
 
EnergyFrequencyWavelength(nm)Type of RadiantEnergyReflectsAbsorbs
>************>************
<0.1gammanon-visible0.1 - 10x-raysnon-visible<380ultra-violetUV380-435435-500500-570570-600600-630630-700visiblegreen-yellowyellowredbluegreen-bluegreenvioletbluegreenyelloworangered>750infra-red (heat)non-visibleIR>25 x 10radiowaves (uwaves)
7
continuousnon-visibleThe energy absorbed is distributed throughout the structure. The absorption pattern of complex organic molecules will be the cumulative sum of the absorption of all of theindividual covalent bonds as a result of the energy's distribution throughout the molecule. As the energy is released from the atoms, molecules, or incandescent solids, an emissionspectra is formed.B.
Emission
– spectra are of 
three
types.1.
Line spectrum
is light of one wavelength formed as the result of excited electronsof 
atoms
returning to their ground state. The
atom
emits the same wavelengthsas those absorbed by the atom.2.
Band spectrum
is a group(s) of wavelengths formed from the return of excitedelectrons in
molecules
returning to their ground state. An array of wavelengths areemitted as the process is complicated by the internal energy of vibration androtation (all substances except rarefied gas and atoms).3.
Continuous spectrum
is a very broad and continuous group of wavelengthsemitted by incandescent solids (tungsten lamps).Electromagnetic radiation in its various forms is measured in common laboratory instruments: A.Absorption of EM radiation1.atomic absorption2.spectrophotometryB.Emission of EM radiation1.flame photomete2.fluoromete3.scintillation counter 
 
UNIT: Electromagnetic Radiation and Photometric Equipment (continued)
MLAB 2401 - Clinical Chemistry Lab Manual
C
B 17
Transmission and Absorbance InstrumentsFluorescence andInstrumentsNephelometersReflectance Spectrophotometers (diagram above left), as found in many modern instrumentssuch as UA dipstick readers and Kodac’s Ectachem: A, light source;B, slit;C, filter or wavelength selector;D, collimating lens or slit;E, test surface;F, collimating lens or slit;G, detector;H, readout device.

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