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8/6/2019 uv vis spec
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AppliedTechnologyDevelopmentDivision
UV/VIS SPECTROPHOTOMETER
MECASYS, Co., Ltd.
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What is UV/VIS?What is UV/VIS?
Electromagnetic Spectrum
X-Rays UV VIS NIR IR Micro wave
200nm 400nm 700nm 2500nm 500 m
E0
E1
E= E1 -E0
Molecular rotationMicro Wave
Molecular vibrationIR
Outer ElectronUV/VIS
Inner ElectronX-Rays
Type of TransitionType of Radiation
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Principle of UV/VIS SPECTROPHOTOMETERPrinciple of UV/VIS SPECTROPHOTOMETER
Principle
Light Intensity Change : By Absorbance or Transmittance
Quantity : Using Absorbance
I
I
Sample
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LambertLambert--BeerBeers Laws Law
Transmittance, t
t / 0 = t
Absorbance, A
log 1 / t = A
Lambert Beers Law
A = abC, A = bC
b : Sample Path length, C : Sample Concentration,
a : Absorbance Constant
: Molecular Absorbance Constant
I0 It
Sample (Conc.C)
Path lengthb
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Deviations in absorptivity coefficients
at high concentrations (>0.01M) due to electrostatic interactions
between molecules in close proximity
Scattering of light
due to particulates in the sample
Fluorescence or Phosphorescence of the sample
Changes in refractive index at high analysis concentration
Shifts in chemical equilibria as a function of concentration
Stray light
Limitation of LambertLimitation of Lambert--BearBears Laws Law
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Application ofApplication ofUV/VIS SpectrophotometerUV/VIS Spectrophotometer
Quantity analysisQuantity analysisQuantity analysisQuantity analysis
(Using a Spectrum)
Quality analysisQuality analysisQuality analysisQuality analysis
(Standard Curve)
Chemical analysis
Food safety analysis
Blood analysis
DNA/RNA Conc. analysis
Residual Pesticide analysis
Residual chlorine analysis
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Functions of SpectrophotometerFunctions of Spectrophotometer
ABS/%T/CONC
Measuring Absorbance/Transmittance/Concentration
Survey Scan
Measuring Absorbance(Transmittance) Spectrum
Standard Curve (Calibration Curve)
Determining Proportional Constant between ABS and Conc
Simple Kinetics
Measuring Time Dependent ABS Variation
ABS Ratio
Measuring ABS Ratio (DNA/Protein)
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Structure of UV/Visible SpectrophotometerStructure of UV/Visible Spectrophotometer
Light
source
Lens
or
Mirror
Monochromator
Detector
Sample
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ExampleExample
Spectrophotometer
Light
SourceMonochromator
Sample
Compartment
Sensing
Part
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Light SourceLight Source
Deuterium Arc Lamp
UV Region Wavelength Range :
190~420nm
Tungsten LampTungsten LampTungsten LampTungsten Lamp Wavelength Range : Part of the UV and the whole of the VisibleWavelength Range : Part of the UV and the whole of the VisibleWavelength Range : Part of the UV and the whole of the VisibleWavelength Range : Part of the UV and the whole of the Visiblerangerangerangerange ((((350 ~ 2,500350 ~ 2,500350 ~ 2,500350 ~ 2,500nm)nm)nm)nm)
Xenon Lamp Wavelength Range : 190~800nm
Tungsten LampTungsten LampTungsten LampTungsten Lamp
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MonochromatorMonochromator
Accepts polychromatic input light from a lamp and outputs
monochromatic light Components : Entrance slit, Dispersion device, Exit slit
Entrance slit
Exit slit
Dispersion device
White Light
Monochromatic
Light
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Dispersion Device (I)Dispersion Device (I)
Filter
Absorptive Filter Interference Filter : FWHM 10
50 nm
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Dispersion Device (II)Dispersion Device (II)
Prism
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Dispersion Device (III)Dispersion Device (III)
Grating
Plane Grating, Concave Grating
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Monochromator structureMonochromator structure
Littrow mounting
Slit
Grating
Mirror
Czerny-turner mounting
Mirror Mirror
Grating Entrance
slit
Exit slit
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Monochromator DesignMonochromator Design
Modified Czerny-Turner Type
- Focal Length : 200mm- Sine Drive Mechanism
- 1nm Bandwidth
Czerny-Turner Type
- Focal Length : 100mm- Direct Drive Mechanism
- 4nm Bandwidth
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Absorption CellAbsorption Cell
Receptacle for Sample.
Material Quartz, Fused Silica(Using over
190nm)
Glass (Using Over Visible Region)
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DetectorDetector
Photodiode
Photodiode & CCD
PMT(Photo Multiplier tube)
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Type ofType ofUV/VIS SpectrophotometerUV/VIS Spectrophotometer
Single Beam Type Double Beam Type
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Type ofType ofUV/VIS SpectrophotometerUV/VIS Spectrophotometer
Grating
Light Sample
Detector
Scanning type : PDA type :
Grating
Light
Multi channel
DetectorSample
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Specifications ExampleSpecifications Example
Spectral Bandwidth : 1 nm
Wavelength Range : 190 1100 nm
Accuracy : 0.5 nm
Reproducibility : 0.1 nm
Photometric Range : -0.3 3.0 Abs (0 125 %T)
Accuracy : < 1% at 1 Abs
Reproducibility : 0.001Abs
Stray Light :
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Accessories (I)Accessories (I)
Multi-Cell Holder Sipper System
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Accessories (II)Accessories (II)
Micro Volume Cell Round Cell Long Path Cell
Temperature Cell Holder, Film/Thin-Layer Holder
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Thank You!