Integrating Peaks from a DAD or UV Chromatogram in Analyst® Software

For research use only. Not for use in diagnostic procedures.

Answer

Analyst® software Quantitate mode can be used to integrate peaks from DAD or UV chromatograms that were acquired during an LC/MS experiment.

Select Build Quantitation Method under Quantitate, and then select the LC/MS data file and the sample to analyze. A quantitation method building screen will appear with three tabs: Components, Integration, and Calibration. On the Components tab, select DAD as the Data Source.


Then begin to list the peaks for integration under the Analytes section. Give each peak to be analyzed at distinctive name and indicate the wavelength to be evaluated for both DAD and single wavelength data. If a peak needs to be evaluated at multiple wavelengths, enter the peak name twice with the different wavelengths indicated as shown below:



Once all the peaks that need to be evaluated have been entered under the Analytes table, select the Integration tab. The DAD chromatogram will appear showing data for the first wavelength indicated in the table above.



Use the cursor to highlight the peak that corresponds to the first peak entered into the table. Then select the Select Peak icon (circled in red above) and then select Apply. The peak of interest will become highlighted in blue, as shown below:



To select a different peak in the chromatogram or the same peak from a different UV wavelength, select the peak from the Analyte drop down menu (shown circled in red below).



Select peak and apply:

Once all peaks have been selected and the integration applied, go to the File menu and select Save As. Name the quantitation method.

After building the method, select Quantitation Wizard, and then select data file and samples. Then select settings and query. On the next screen, specify the quantitation method to be used, and then select Finish. A results table will appear, but the integration values for the DAD peaks will not be showing. Right click on the table to bring up the settings menu and select Table Settings. Select the folder for Columns, and a Results Table Columns screen appears. Select Analyte from the drop down menu, and then select Analyte Peak Name, Analyte Peak Area for DAD, and Analyte Peak Height for DAD. Then select Done. These columns with the integrated peak areas from the DAD data will now appear in the results table.