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Uv Food Ana
Uv Food Ana
INTRODUCTION
of solutions, transparent or opaque solids, such as polished glass, or gases. The instrument
operates by passing a beam of light through a sample and measuring the intensity of light
reaching a detector. When a photon encounters an analyte molecule (the analyte is the
molecule being studied), there is a chance the analyte will absorb the photon. This absorption
reduces the number of photons in the beam of light, thereby reducing the intensity of the light
beam. Click on the Start button to start the simulation and the Stop button to stop the
simulation. The light source is set to emit 10 photons per second. The intensity of the light
reaching the detector is less than the intensity emitted by the light source.
However they can also be designed to measure the diffusivity on any of the listed light ranges
that usually cover around 200nm - 2500nm using different controls and calibrations. Within
these ranges of light, calibrations are needed on the machine using standards that vary in type
solution. A certain chemical reaction within a solution may occur in a forward and reverse
direction where reactants form products and products break down into reactants. At some
point, this chemical reaction will reach a point of balance called an equilibrium point. In order
to determine the respective concentrations of reactants and products at this point, the light
transmittance of the solution can be tested using spectrophotometry. The amount of light that
passes through the solution is indicative of the concentration of certain chemicals that do not
PROCEDURE
1. (200 X 10-4 g/100ml ) of dye stock solution ( erythrosine ) was diluted into a series of
different concentration as below :
BLANK:
5 X 10-4 g/100ml
10 X 10-4 g/100ml
15 X 10-4 g/100ml
20 X 10-4 g/100ml
25 X 10-4 g/100ml
allow it to warm up .
2. The cuvette is filling about 2/3 full with the blank solution and another with 10 X 10-4
3. The uv visible absorbance spectrum for erythrosine was determined .the range of
Spectrophotometer type:
2. The absorbance for each standard solution and the unknown solution was measured.
CALCULATION
M1V1=M2V2
a. 5 X 10^-4 g/100 ml
( 200 )( V1 ) = ( 5 )( 100 )
V1 = 2.5 ml
b. 10 X 10^-4 g/100 ml
( 200 )( V1 ) = ( 10 )( 100 )
V1 = 5.0 ml
c. 15 X 10^-4 g/100 ml
( 200 )( V1 ) = ( 15 )( 100 )
V1 = 7.5 ml
d. 20 X 10^-4 g/100 ml
( 200 )( V1 ) = ( 20 )( 100 )
V1 = 10.0 ml
e. 25 X 10^-4 g/100 ml
( 200 )( V1 ) = ( 25 )( 100 )
V1 = 12.5 ml
GRAPH SINGLE-BEAM SPECTROPHOMETER ABSORBANCE
Concentration Vs Absorbance
0.16
0.14 R = 0.9721
0.12
Absorbance
0.1
0.08
0.06
0.04
0.02
0
0 2 4 6 8 10 12 14
Concentration (mL)
Concentration VS Absorbance
0.14
0.12 R = 0.1297
0.1
Absorbance
0.08
0.06
0.04
0.02
0
0 2 4 6 8 10 12 14
Concentration (mL)
DISCUSSION
In this experiment, we have learned how to determine the erythrosine concentration by using
must be freshly prepared and must be prepared with extreme care because the accuracy of
analyte determination is depending on the accuracy of the standard. As the mineral elements
concentration in foods are at trace level, thus it is essential to use highly pure chemical
reagent and water for preparation of samples and standard solutions. Thats why the deionized
measuring the transmittance of the sample and solvent at each wavelength. Single-Beam
the UV-Vis spectral region. The concentration of an analyte in solution can be determined by
measuring the absorbance at a single wavelength and applying the Beer-Lambert Law. Single-
beam spectrophotometers can utilize a fixed wavelength light source or a continuous source.
The simplest instruments use a single-wavelength light source, such as a light-emitting diode
(LED), a sample container, and a photodiode detector. Instruments with a continuous source
have a dispersing element and aperture or slit to select a single wavelength before the light
The double-beam design greatly simplifies this process by measuring the transmittance of the
sample and solvent simultaneously. The detection electronics can then manipulate the
measurements to give the absorbance. The dual-beam design greatly simplifies this process by
simultaneously measuring P and Po of the sample and reference cells, respectively. Most
spectrometers use a mirrored rotating chopper wheel to alternately direct the light beam
wavelength.
The linearity of the Beer-Lambert law is limited by chemical and instrumental factors. Causes
the sample. Next, changes in refractive index at high analyte concentration, shifts in chemical
be minimized by using a relatively flat part of the absorption spectrum such as the maximum
From our result it show that the higher the concentration, the higher the absorbance value.
spectrophotometer that is 0.1297.In double-beam, error has occur because it suppose the
higher the concentration, the higher the absorbance value. But, start at 10mL, the absobance
value decrease. The error came from Spectrophometer (Perkin-Elmer Lambda 35).
R^2 gives a formula for the line most closely matching those point. It also gives an R^2 value
to say how well the resulting line matches the original data points. Single-beam
spectrophotometer only have a single beam from the light source. The reference standard is
measure to standardize the instrument. For single-beam configuration to perform well, the
light source, detector and electronics must be reasonably stable over time.
The cuvette with erythrosine that is used must be placed in the right slot. The clear side must
face the light source in order to allow the light fully penetrate the sample. While handling the
cuvette, we must not let our fingers to touch the clear side or the light will just penetrate our
The advantage of using the single-beam spectrophotometer is that there are fewer moving part
which make it easier to handle. The beam from the light source of double-beam
spectrophotometer is split into two. One beam illuminates the reference standard while the
other one illuminates the sample. The beam recombined before they reach the single
monochromator. The cuvette containing the reference standard and sample can be placed
together simultaneously but the result obtained will be a bit unorganized and might be
Lastly the precaution step is the cuvette is put one by one. Although it takes longer time, but
the result obtained is more organized and accurate. Double-beam spectrophotometer is more
complicated compare to single-beam because there parts that are not static and moveable. So
each time the parts is removed, it has to be tide back correctly to its place or the reading
CONCLUSION
At the end we know the graph absorbance versus concentration show a linear perpendicular
graphs. As concentration increase absorbance also increase. Result show, the higher the
concentration the higher the absorbance. Comparison between single and double beam have
1) www.chm.davidson.edu/vce/spectrophotometry/Spectrophotometry.html
2) http://www.files.chem.vt.edu/chem-ed/spec/uv-vis/singlebeam.html
3) http://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/Spectrpy/UV-
Vis/spectrum.htm
4) http://elchem.kaist.ac.kr/vt/chem-ed/spec/beerslaw.htm
5) http://www.files.chem.vt.edu/chem-ed/spec/uv-vis/uv-vis.html
6) http://www.chemguide.co.uk/analysis/uvvisible/spectrometer.html