Standard Operating Procedure

For PE Lambda 2 UV-VIS Spectrophotometer

Load UV WinLab Software:
From Windows, double click on the WinLab icon.
Methods:
UV WinLab offers different method types:
Method

Description

Scan

For scanning spectra. Scans a spectrum over a

preset wavelength range.

Time Drive

For ordinate value measurement over a time

period, e.g. for kinetics. Measures the ordinate
values of a sample over a time period.

Wave Prog.

For ordinate value measurement at different

wavelengths, and calculating differences and
ratios of these ordinate values. Measures the
ordinate values of a sample at several
wavelengths, and calculates ratios, or differences
of these values.

Conc.

For concentration measurement and calibration

curves. Measures the concentration of a
component in a sample. With Concentration
methods a calibration curve is first established,
followed by measurement of the sample.

To Access Methods:
You can access the various method types via the Applications menu or via the
method editor icons in the toolbar. You can open specific prepared methods for
the Methods window or via the File menu.
The Setup Button:
Sets the spectrometer to the required starting conditions entered on the parameter
pages of the respective method. The button does not show when the spectrometer
conditions already correspond to those of the method. Once the spectrometer is

setup, the Start button turns green to indicate that the spectrometer is ready to
start.
The Autozero Button:
Sets the baseline to zero. The Autozero button turns to a red Stop AZ button
when activated. When you click on the Stop AZ button, the spectrometer and the
method return to the start conditions.
The Start Button:
Starts the corresponding method. The Start button turns to a red Stop button
when activated. When you click on the Stop button, the spectrometer and the
method return to the start conditions. If the Start button was activated before the
Setup button (i.e. not green), the text dims to gray, the spectrometer is set up to
the required start conditions and then the method starts. This may take several
seconds before the method is started, and is therefore not recommended for
kinetic methods.
Scan, Time Drive(Td), Wavelength Program(Wp), or Concentration(Conc):
A. To create or open a method:
1. In the Methods window, double-click on the desired method file.
...or...
In the toolbar, click on the Scan, Time Drive, Wavelength Program, or
Concentration icon.
...or...
In the menu bar, display Applications and select Scan, Time Drive, Wav.
Prog., or Conc.
...or...
In the menu bar, display File, select Open and open the required method.
2. Click on the tabs at the bottom of the method editor window to open the
various parameter pages.
3. Set the parameters on each page as required.
4. Once all the parameters are properly set, click on Setup. The new
parameters are set on the instrument. The Start button turns green to
indicate the spectrometer is ready.
B. To perform Autozero:
1. Open the sample compartment cover.
2. Insert matched cells into the sample and reference cell holders.
3. Close the sample compartment cover.

4. In the toolbar, click on Autozero. Autozero (background correction) is

performed. The Autozero button turns to a red Stop AZ button.
C. To measure the sample (Scan, Time Drive or Wavelength Program):
1. Open the sample compartment cover. Remove the blank from the sample
cell holder.
2. Insert the sample cell in the sample cell holder.
3. Close the sample compartment cover.
4. In the toolbar, click on Start. The spectrometer starts the scan. The Start
button turns to a red Stop button. A graph window or results window
opens showing the scan/results.
5. Repeat for each sample
D. To establish a calibration curve (Concentration only):
1. Open the sample compartment cover. Remove the blank from the sample
cell holder.
2. Insert the first reference solution cell in the sample cell holder.
3. Close the sample compartment cover.
4. In the toolbar, click on Start. The spectrometer starts the calibration. The
Start button turns to a red Stop button. Graph and Results windows open
on the screen to show the calibration data.
5. Insert the other reference solutions as requested by the software.
6. Accept or change the calibration data as required. A Conc-Calib dialog
box appears with a selection of buttons to allow you to do this.
E. To measure the sample (Concentration only):
1. Open the sample compartment cover. If necessary, remove the blank or
reference from the sample cell holder.
2. Insert the sample cell in the sample cell holder.
3. Close the sample compartment cover.
4. In the Conc-Calib dialog box, click on Measure Samples. The
spectrometer starts the concentration method. The Start button turns to a
red Stop button. Data is shown on the Graph and Results windows.

5. Insert the samples as requested by the software. The results are shown on
the Results window, and spectra on the Graph window.
*To take repeated readings at a single wavelength:
Click on Application and then select Manual. Set the Wavelength to the
value you require and close the dialog box.
F. To Save Data:
To save the data from a Results window:
1. Select the window that contains the data to save.
2. c. This saves the data with the same filename.
...or...
In the menu bar, display File and select Save As. This saves the data
with a different filename.
To save the data automatically after data collection:
1. On the method page, select Autosave.
G. To Print:
Select the item you want to print (window, file).
In the menu bar, display File and select Print.
To print out the data automatically after data collection:
1. On the method page, select Autoprint.