How to perform a scan on the ACQUITY FLR using the ACQUITY UPLC console - WKB2892

OBJECTIVE or GOAL

Determine the optimal excitation and emission wavelengths using the ACQUITY Console, by performing a scan on the ACQUITY FLR.

ENVIRONMENT

• ACQUITY FLR
• Empower 3

PROCEDURE

1. Set the gain on the ACQUITY FLR detector to zero.

• In the ACQUITY UPLC console, navigate to the interactive display
• Select the control icon.
• Once the gain setting link turns blue, click on the value "0", and then click Enter.

2. Remove the flow cell.

3. Install the cuvette cell.

4. Perform a zero scan:

• Remove the cuvette holder from the cuvette cell
• Fill one of the cuvettes with sample diluent and cap the cuvette
• Install the cuvette in the holder, making sure its bottom is flush with the bottom of the groove in the cuvette holder
• Reinstall the cuvette holder in the cuvette cell
• Initiate the scan function: Navigate to the FLR page in the console and select Control > Scan wavelengths
• Select the following parameters
  • Zero Scan
  • The grating to be scanned
  • Scan rate (default is 100 nm/min)
  • Range to be scanned
  • Stationary grating (this is the grating not being scanned)
  • Fixed wavelength of the stationary grating Rule: This wavelength must not be within the scanned range. It must be at least 10 nm from the lowest wavelength (emission scan) or the highest wavelength (excitation scan)
  • Select an appropriate gain setting

5. Once all parameters are set, click Start to initiate the scan.

6. Click Results to view the zero scan; a spectrum window appears in the bottom-left portion of the window.

7. Perform a sample scan:

• Remove the cuvette holder from the cuvette cell and remove the sample diluent cuvette
• Fill the second cuvette with sample and cap the cuvette
• Install the cuvette in the holder, making sure its bottom is flush with the bottom of the groove in the cuvette holder
• Reinstall the cuvette holder in the cuvette cell
• Select Sample Scan; Tip: Do not change any other settings or it deletes the zero scan from memory

8. Click Start to initiate the sample scan.

9. The sample spectrum is displayed in the Results window and the sample scan peaks are displayed in the table. The four most intense peaks are recorded in the table.

10. Repeat the zero and sample scans to obtain the other maximum for the excitation/emission pair.

ADDITIONAL INFORMATION