**Liquid Chromatography Operating Procedures**
Liquid chromatography is a widely used analytical technique for separating, identifying, and quantifying compounds in a mixture. Proper operation of the liquid chromatography system ensures accurate and reliable results. Below is a detailed guide to operating the system.
**1. Power On Sequence**
Before starting the instrument, ensure that all components are properly connected. Turn on the power in the following order:
- Pump
- Detector
- Computer host
- Monitor
- Printer
**2. Launching the Chromatography Workstation Software**
Double-click the "Chromatography Workstation" icon on the desktop. Enter the password “SAGE†and click OK. Then, select the “Alltech-426†icon to begin the session.
**3. Setting Chromatographic Conditions**
- **Pump Settings**: Adjust the flow rate using the arrow keys. Press RUN/STOP to start the pump.
- **Detector Settings**: After the detector passes its self-test, set the required wavelength. Use the ZERO button to reset the baseline before analysis.
**4. Preparing the Mobile Phase**
Mix the mobile phase according to the desired volume ratio. Ensure thorough mixing, then filter and sonicate for 20 minutes to remove any air bubbles.
**5. System Preparation**
- Insert the filter into the mobile phase reservoir.
- Open the bypass valve on the pump and press the PURGE button. Use a syringe to draw and flush the mobile phase until no air bubbles remain in the tubing.
- Press RUN/STOP to activate the pump.
**6. Sample Injection and Analysis**
- On the computer, click the blue “RUN/SINGLE†icon. Rename the data file (e.g., SAM01.001) and click OK.
- Load the injection valve to the LOAD position. Inject the sample using a microliter syringe. Press the detector’s ZERO key, then move the injection valve to the INJECT position and press the spacebar on the keyboard. The system will begin analysis.
**7. Data Collection and Reporting**
- Once the chromatogram is complete, the system stops automatically. Click the “Analyze†icon to calculate peak areas.
- Select “Reports†and choose “Area%†to view the results. To print, click the printer icon, select “Print Report P,†and choose “Custom C.â€
**8. Repeating the Process**
Repeat the injection and analysis steps for each sample. Allow about 5 minutes between samples for system stability.
**9. Shutting Down the System**
Before turning off the system:
- Flush the column with the mobile phase for 20–30 minutes. If the column will not be used for an extended period, rinse it with pure acetonitrile.
- Turn off the pump, detector, monitor, and computer in reverse order. Always exit the software before powering down.
**Notes:**
- Never turn off the computer while the software is running.
- The pump flow rate should not exceed 1.5 mL/min under normal conditions.
- The pump will not start if the mobile phase is depleted.
**Modifying Analysis Conditions**
- **Adjusting Run Time**: Go to “Setup,†set the sampling frequency to 2, and define the run time (e.g., 10 or 20 minutes). Save the method after setting the channel status and trigger options.
- **Integral Parameters**: Modify the Minimun Area, Width, and Threshold values in the Integral Parameter Table to refine peak detection.
- **Report Format**: Customize the report by selecting fields like File Name, User Name, Date, and Peak Information. Set column widths and decimal places as needed.
- **Accessing Previous Methods**: Use the Method and Data icons to load previous methods or data files.
**Internal Standard Method**
- Select the appropriate method and enter the standard.
- After recording the spectrum, adjust the calibration settings and input the internal standard concentration.
- Recalculate and verify the correction factor. Repeat for unknown samples and print the results.
**External Standard Method**
- Follow similar steps as the internal standard method, but use external standards for calibration.
- Input the standard concentrations and verify the calibration curve. Calculate the standard deviation and proceed with sample analysis.
**Company Information**
Shanghai Qite Analytical Instrument Co., Ltd.
Website: http://www.qite.com.cn
Contact: Tel/Fax: +86-21-XXXXXXX
For more information, visit: http://news.chinawj.com.cn
This guide provides a comprehensive overview of liquid chromatography operations. Following these procedures ensures consistent and accurate results. Always refer to the manufacturer's instructions for specific details related to your equipment.
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