GUCThe German University in Cairo
Instrumental Analysis I
WS2015 - PHCMp561
Technical notes on the use of the UV-Visible Spectrophotometer
1. Turn on the power to the Instrument JASCO V-630.
2. Turn on the computer and monitor.
3. Double click the ‘’ JASCO Spectra manager ‘’ icon located on desktop. Fig 1
Fig 1
4. You can choose between ‘’Instrument icon’’ or ‘’Analysis icon’’. Fig 2
5. You can choose from different 8 modes of measurements.
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
I.
Instrumental Analysis I
WS2015 - PHCMp561
Fixed wavelength measurement (for construction of calibration curves)
1- Set parameters of Measurements
 Click [Measure] – Parameters Fig 3
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Fig 3
On the [General] tab, write the appropriate λ of measurement as mentioned in
your experiment. Fig 4
Fig 4
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
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Instrumental Analysis I
WS2015 - PHCMp561
Before doing any measurement you have to [Auto Zero ] the spectrophotometer
(this sets absorbance value to Zero at this current wavelength)
Make sure no cuvettes are placed inside the cell (Cuvette) holder, only air.
Click on [Control] – [Auto Zero]. Fig 5
3-Measuring absorbance of standard solutions and unknown samples
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Place your Blank into the Blank cuvette holder.
Place the standard/sample solutions in one of the cuvette holder
Arrange your samples in ascending order of their concentration
Make sure you keep record of the position of each concentration within the cuvette holder
Select [Measure] - [Sample]. Fig 6 (or click button; Fig 7)
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
Instrumental Analysis I
WS2015 - PHCMp561
Fig 6
Fig 7
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
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Instrumental Analysis I
WS2015 - PHCMp561
Copy your absorbance values from the result worksheet to your lab report. Fig 8
Plot calibration curve using Microsoft Excel as instructed in your lab report.
Fig 8
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
Instrumental Analysis I
WS2015 - PHCMp561
II. Spectra Measurement
1. Choose from the instrument menu the [Spectra measurement] mode.
2. A spectral scan screen will be displayed showing an absorbance spectrum.
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
Instrumental Analysis I
WS2015 - PHCMp561
1- Set parameters of Measurements
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Click [Measure] – Parameters
On the [General] tab adjust the wavelength range that you need for the scanning
process (ex. 200-400 nm)
The data interval is adjusted to 5nm
Use continuous scanning mode
2- Measuring absorbance of the baseline
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Before doing any measurement you have to [Auto Zero ] the spectrophotometer (this sets
absorbance value to Zero at this current wavelength)
P.S. Make sure no cuvettes are placed inside the cell (Cuvette) holder, only air
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Click on [Control] – [Auto Zero].
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Select [Measure] - [Baseline] to adjust the baseline to zero.
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
Instrumental Analysis I
WS2015 - PHCMp561
3- Scanning a standard solution
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Insert one cuvettes (Quartz or plastic depending on the λ of measurement)
containing the blank into the blank cell holder and another one containing the
standard/sample into sample cell holder
Click on [Measure] - [Sample].
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Spectral scan will be performed and spectra analysis window pops up.
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
Instrumental Analysis I
WS2015 - PHCMp561
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Click on File save data to save the performed spectrum.
To find the λ max of the spectrum see the following figure
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The following window appears
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From the parameters window choose Peak [Both]
Noise level[0.01]
Then click [apply]
The different λ in the spectrum will be displayed on the left of the screen.
Student guide JASCO V-630
,,,
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
Instrumental Analysis I
WS2015 - PHCMp561
3- Comparing Spectrum of two standard solutions
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Perform a spectral scan as mentioned above
Save your spectral scan
Perform a new spectral scan for the new substance
Wait until the new spectra analysis window pops up
Choose [File] [overlay]
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Select the needed spectrum from the saved data
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
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Instrumental Analysis I
WS2015 - PHCMp561
Activate the required spectrum by clicking on view 2 .
Click on [Processing] [Peak Processing] [Peak Find]
Student guide JASCO V-630
Nermin Salah, Nayra Omran
GUCThe German University in Cairo
Instrumental Analysis I
WS2015 - PHCMp561
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Click on apply
The different λ will be displayed on the spectrum
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Determine the λ from the spectrum overlay based on the criteria specified in your lab
manual.
Student guide JASCO V-630
Nermin Salah, Nayra Omran