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Quick Start Guide To Shimadzu UV

This document provides a quick start guide for using the Shimadzu UV-2450 spectrophotometer. It outlines the startup process and describes the basic functions and modules for collecting absorbance data, creating standard curves, and performing kinetic analyses. Instructions are provided for baseline correction, peak picking, area calculations, data manipulation, and generating reports.

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719 views

Quick Start Guide To Shimadzu UV

This document provides a quick start guide for using the Shimadzu UV-2450 spectrophotometer. It outlines the startup process and describes the basic functions and modules for collecting absorbance data, creating standard curves, and performing kinetic analyses. Instructions are provided for baseline correction, peak picking, area calculations, data manipulation, and generating reports.

Uploaded by

chemchemha
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Quick Start Guide to Shimadzu UV-2450

1. Login to the computer with your UVM netid and password.


2. Turn on the instrument, making sure the software is not open. Allow 20 – 30 minutes
for the lamps to warm up.
3. Open the UV Probe software from the desktop.
4. Go to the Window menu and pick a module (1,2 or 3). The module options are
spectrum, photometric and kinetic. Spectrum is suitable for wavelength range scans.
5. Click Connect on the bottom of the window to begin communication with the
spectrophotometer. The connection and testing process will take a few minutes. Be
patient. If the integrating sphere is in the sample compartment the deuterium lamp
may fail. This is OK.
6. Create a data collection method – specific to each module, see below.

Spectrum Module – absorbance, transmittance, reflectance or energy readings scanned


through a range of wavelengths.

1. Select Edit > Method to specify data collection parameters.


a. Enter the initial and final wavelengths to be scanned, scan speed, sampling
intervals, and any repetitions.
b. Click the Instrument Parameters tab and input a measuring mode, slit width and
light source change wavelength. Make sure that the S/R Exchange is set to
normal.
c. Click OK and then save your method by going to File > Save As and naming the
file as you wish. Choose Method File (*smd) and then click Save. If no folder
exists for you, please create a new folder with your last name and store all data
therein.
2. Make sure that there is nothing in the sample compartment. Next, click Baseline on the
button bar to set the background of your spectral region of interest to zero by
performing a baseline correction.
a. In the dialog box, enter the wavelengths you wish to scan; this should be the
same region as the one specified in your recently created method. Click OK.
i. Look in the lower left hand corner for instrument status and be patient.
b. To verify that the baseline measurement was stored, click on the Instrument
History tab on the Output Window and see that it is listed.
3. You will need to configure the graph before you acquire data. Click the Overlay tab.
Click the minimum absorbance value on the Y-axis and change it to -1. Click the
maximum absorbance value on the Y-axis and change it to 3.
4. Place your sample in the proper cuvette holder. Click the Start button on the
Instrument bar to initiate data collection. The scan will take a few minutes. When the
scan is complete a New Data Set dialog box will pop up.
a. Enter the file name in the New Data Set dialog box that appears. Click OK.
b. Now the data is saved in memory only and not to disk. If you close the
program, you will lose your data.
c. Save the data by selecting File > Save As. Select the appropriate directory for
your data and enter the file name. Select .spc in the Save As Type list. Click
Save.
d. To view the data set parameters, double click on the data set in the Legend
Window.
5. To pick peaks, first right click on the graph and choose Auto Scale to display all of the
data. Each peak and valley should now be numbered.
a. Select Operations > Peak Pick. If necessary, use the View menu to hide the
Method pane and Output window to create more room onscreen.
b. To remove numbers from peaks or valleys, right click on the Peak Pick table and
select Mark Peaks or Mark Valleys as desired.
c. To adjust the Peak Pick threshold value, right click on the Peak Pick table and
select Properties. Click on the thumb tack to pin the dialog box and adjust the
values as appropriate.
6. To calculate the peak area, you must first define the region of interest by selecting
Operations > Peak Area. Select Peak Area table, click the Start column for Region 1.
a. Enter the starting and ending wavelengths of the peak(s) of interest and press
Enter to define the peak area region.
7. To manipulate a data set, select Operations > Manipulate. Pick the type of
manipulation you desire (ex. Arithmetic) and select the proper data set.
a. In the Operation list select the subtype manipulation (ex. Subtraction) and
populate any other desired fields.
b. Enter any notes in the New Data Set dialog box.
c. Click the Stacked tab in the Graph pane to display both the raw and manipulated
data sets.
8. To build a report, first close the Instrument bar, Photometer status bar and the Output
Window (go to View and uncheck). Ensure that all toolbars are displayed by going to
the View menu.
a. Go to Window > Spectrum. Right click on the spectrum and click Auto Scale.
Then go to Edit > Copy. Go to Window > Report Generator and select Edit >
Paste. Drag the corner to resize the graph. Look at the last set of buttons on the
left side toolbar. These are options to add to your report from Spectrum mode.
b. Create a report title by clicking on the Text Object icon. Enter the desired title in
the text box. Position the text box where you would like it.
c. Right click the text object and select Properties. Click the Fonts tab, and choose
your Font settings.
d. When your report has been configured, select File > Print Preview to view the
report as it will appear.
i. If unsatisfied, click Close to return to the Report Generator.
ii. If satisfied, click Print. Select HP Laserjet 4L. Click OK.
e. Save the report go selecting File > Save As and entering the file name. Make sure
to save in the directory with your last name under C: > UVVis data. If no folder
exists, please create one.
Before you end your session, be sure to transfer your files to either your zoo
account or active directory (see details below). Data files will be wiped once a
semester.
i. Go to My Computer and locate the directory called “winhome”; this will
save directly to your folder on active directory:
\\files.campus.ad.uvm.edu\netid.
ii. Your active directory folder is available from anywhere on campus. To
access this folder from a PC, click Start > Run… and type the address
underlined above. From a Mac click Go > Connect to Server and type the
address to access the drive.

When finished, close the program and log off the computer. Press control + alt + delete and
select Log Off.

Photometric Module – determine concentrations, create standard curves and derive values
from customized equations. See chapter 3 in the instruction manual for a tutorial.

Kinetic Module – time dependent sample changes in absorbance, transmittance,


reflectance or energy. See chapter 4 in the instruction manual for a tutorial.

Notes about data acquisition:

Slower scan speeds result in higher resolution. Medium speed is usually sufficient.
Typical slit width settings are 1.0 and 2.0 nm.
Full range of instrument is 1100 nm – 190 nm.
Light source change wavelength may be set anywhere from 364 – 295 nm. Choose a
region that won’t interfere with your spectrum.
We have no control over the PMT gain.

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