Read this manual thoroughly before using the product and use the product in accordance with the instructions in this manual. If the user or usage location changes, ensure that this manual is always kept with the product.
LC-40D XR CL, LC-40D X3 CL, or LC-40B X3 CL
SIL-40C XR CL or SIL-40C X3 CL Autosampler
CTO-40C CL Column Oven
SPD-40 CL UV-VIS Detector
Optional Parts
Can be connected to up to six sets of 1L bottle holders or up to four sets of large bottle holders. Used in combination with an earthquake system anchor strap, this kit can be used to secure the system to the base stand.
Configuration List
Switching ON the Instruments Power Supply
When the main power switch is switched from OFF (○) to ON ( ), the instrument starts without pressing the (power) button. If an optional PO-6P socket unit is used, the main power can be switched on/off from the front of the instruments.
Logging in to LabSolutions and Opening the [Realtime
Displays the menu choices enabled for the current window, based on the user permissions for the currently logged in user.
Specifying Analytical Conditions
Editing Methods
Clicking this button sets the stop time for all detectors at the same time as the [LC Stop Time] setting. Pump gradient programs can be specified by changing the pump mode to high pressure gradient mode.
Placing Mobile Phases and Autosampler Rinse
Placing Mobile Phase and Rinse Solution Bottles on the
Parameters for monitoring the levels of the remaining mobile phase and the rinse solution can be set by clicking [Control Panel]-[Mobile Phase Settings]. If the optional mobile phase level monitor is used, the remaining level is monitored in real time using a scale, eliminating the need to reset the volume each time you place the mobile phase bottle.
Performing Auto-Purging
Manual Purging the Solvent Delivery Pump
1 Insert the suction tube for the flow channel to be flushed into the reservoir bottle containing the mobile phase and place the bottle on the reservoir tray. Continue automatically purging each manually flushed mobile phase port until all bubbles have cleared from the flow channel.
Attaching the Column
Attaching Columns in the Column Oven
In such cases, turn the cell temperature control OFF and wait for the temperature to stabilize before proceeding with the analysis (approximately 60 minutes). Disabling cell temperature control can increase the range of baseline oscillation in response to room temperature fluctuations.
Warming Up the System
Specify the date and time when booting should begin, select the created method file, and select the [Run LC Autopurge] check box. If the [Perform LC Autopurge] check box is selected, the automatic purge and warm-up actions specified in the selected method begin.
Placing Samples
Preparing Sample Vials (or Sample Plates)
For sample vials with a separate septum and cap, attach the cap with the PTFE side of the silicone septum facing down (toward the liquid). If samples containing a volatile acid are used, be sure to cap the sample vials.
Placing Samples in the Autosampler
Attempting to remove the sample rack when the status indicator is flashing amber will pause injection operations. The temperature control function automatically restarts when the sample rack is returned to its correct position.
Performing a Single Run Analysis
Performing a Single Run Analysis
Specifies the number indicating the position of the sample well and the number indicating the position of the wells filled with samples. Running” is displayed at the top of the main analysis window so that the analysis status can be confirmed.
Performing Postrun Analysis
Retention times can be entered automatically by clicking on a peak in the [Chromatogam View] with the [Ret. By using the method settings specified here for the rest of the analysis, data analysis results can be checked immediately after the analysis is complete.
Displaying and Printing Data Reports
Performing Realtime Batch (Sequence) Analysis
Performing Realtime Batch (Sequence) Analysis
Click the vial number to start the analysis and drag the cursor to the vial number to end the analysis to specify the starting vial number, the ending vial number, and the number of samples. This setting is related to the [Sample Type] setting on the left side of [Batch Setting].
Analyzing Multiple Sets of Data Using Data Browser
4 Drag the batch file containing the data to be analyzed into the [Quantitative Results View]. To add parameters to display, right-click the [Quantitative Results View] and select [Table Style].
Outputting Summary Reports
To use a different report format, drag and drop the report format file onto the report. Double-clicking on an item in the report displays the [Properties] window, where you can specify the display settings.
Stopping Analysis and Shutting OFF the Power
Check List to Confirm after Analysis is Finished
Purging Columns with a Column Storage Solution
Shutting Down the Instruments
Setting Up Startup for Next Use
Key Points for Analysis
Before Analysis
Use an autosampler rinse solution that is the same as the mobile phase or has a similar composition. 7 Check that the flushing solution drains from the end of the FEP tube at the outlet end of the automatic seal flushing kit.
Precautions for Selecting and Handling Mobile Phase
Precautions When Replacing the Mobile Phase
If the status indicator is yellow without flashing, the sample rack can be removed even during analysis. Injection operations will not start if a sample plate with samples to be analyzed is not placed on the sample rack.
Precautions for Selecting an Autosampler Rinse Solution
Bubbles can sometimes form when you switch the rinse solution from water to an organic solvent. After the analysis is completed, flush the channels to replace the flushing solution with water or methanol.
Using Buffer Solutions as a Mobile Phase
If a high-concentration buffer solution and an organic solvent are used for low-pressure gradient analysis, the tubing must be changed between. The startup function is used to automate processes ranging from preparation to analysis at a specific time to equilibration of columns by heating the system.
Startup Function
Executing the Startup Process
If the system power is OFF at the time specified by the startup function, the system is automatically powered on. By specifying the startup function within batch settings, batch analyzes can be started automatically after the startup function is executed.
Executing a Startup Process and Starting a Batch
Shutdown Function
Executing the Shutdown Function
1 Shutdown Method File Specifies the method file to use when shutting down the system. 2 Cooldown Time Specifies how long to operate the system before performing the shutdown process.
Executing a Batch File with a Shutdown Function
Sets whether or not to execute the cooldown process using the specified method file when the shutdown process is executed. 4 Cool-down time Specifies how long the system must operate before performing the shutdown process.
Auto-Diagnostics (Pulsation Check) and
Auto-Diagnostics (Pulsation Check)
1 Click the (Realtime Batch) icon on the [main] assistant bar in the LabSolutions [Realtime Analysis] program. 4 Click the (Start Real-time Batch) icon on the [Real-time Batch] assistant bar to start the batch analysis.
Autosampler Functions
Pretreatment Functions
Such peak shapes can be improved by simultaneously injecting the sample with a solution containing an ion pair reagent. Samples can be detected from multiple peaks in the same data file by sequentially injecting the same sample.
Cut off Loop Function
Low Dispersion Injection
Batch Action Functions
Stop (Run Next Queue) Stops the current batch process and executes the next batch process in the batch queue. 2 Enter the row number of the last row in the batch table in the [Parameter] cell.
Using ACTO (Analytical Condition Transfer and
Concentration Gradient Start Adjustment Function
The start time of the concentration gradient program is shifted prior to injection, in proportion to the volume specified in [Adjustment Volume]. The start time of the concentration gradient program is delayed after injection, in proportion to the volume specified in [Adjustment Volume].
Method Migration Functions
Select the [Adjust the gradient start] check box to adjust the gradient delay range differences of the corresponding systems. To use a time program to adjust the concentration gradient delay time, select the [Gradient Program] check box in [How to Adjust].
Useful Data Analysis Functions
Column Management
Column Management Workflow
Setup for LabSolutions
After clearing the CMD information, set the column information again." click the [OK] button. CMD initialization is complete when the [CMD 1 (Oven)] display changes from [Previous Version and CMD-Instrument Inconsistency] to [Not Registered].
Column Registration and Replacement (Discard)
Analysis with LabSolutions
Viewing the Acquisition Log
If you have used the column (CMD) with another instrument, [Column] is automatically removed from [Modules used for analysis] when the [Realtime Analysis] program starts. When a link beep sounds, the column information is reflected and the [CMD-Method Column Mismatch] status changes to something like [Ready].
Hardware Validation Classifications
Hardware validation is the process of verifying and documenting that the analytical tools used for analysis function properly and can be used for the intended analysis. Compared to the various other types of validation related to analysis, such as analytical method validation and system suitability testing, hardware validation serves as a prerequisite for and is more important than the other types of validation.
Timing for Hardware Validation
Periodic Inspections
Given that the performance of the analytical instrument may change over time, due to the deterioration of consumables or components, the equipment should be validated periodically between the time it is inserted and disposed of. This is a contract under which a qualified engineer approved by Shimadzu performs periodic component and system verification and provides results reports.
Routine Inspections
Inspections during Maintenance
Precautions for Periodic Inspections
Parts Required for Periodic Inspections
Used to verify the accuracy of temperature settings on the column oven, autosampler and other instruments. Used to verify the wavelength accuracy of absorption detector and inspect the system (reproducibility, injection volume linearity, transmission and absorption linearity).
Periodic Inspection of Solvent Delivery Pumps
Inspection Item
Testing Flowrate Accuracy
8 Use the procedures listed below to measure the flow rate accuracy of each measurement method specified in step 6. 1 When ready to measure, press [Enter] again and measure the time required for 5 mL to collect into the measuring flask.
Periodic Inspection of Autosamplers
- Inspection Item
- Injection Volume Accuracy Test (Total Injection Method
- Inspection Item
- Checking Temperature Accuracy
Immediately before closing the door, ensure that the status indicator on the plate stack remains lit green. Next, confirm that the status indicator on the disc stack is green, and then perform the procedure in this step.
Periodic Inspection of Column Ovens
Inspection Item
Testing the Temperature Adjustment Function
Attach the thermocouple of the temperature gauge (A) used for the measurements to the column support with 1-cm square aluminum tape (B). 7 Press the [↓] (down arrow) key repeatedly to place the cursor on [VALIDATION], and then press the [→] (right arrow) key.
Periodic Inspection of UV Absorbance
Inspection Item
Wavelength Accuracy Test
1 Confirm that the absorbance has stabilized and then, with the flow cell filled with water, set the wavelength to 200 nm and record the absorbance at that point. 3 Confirm that the absorbance has stabilized and then scan with the flow cell filled with water.
Periodic Inspections of Systems
- Inspection Item
- Absorbance Linearity Test
- Noise/Drift Test
- Gradient Concentration Test (Gradient System)
- Reproducibility Test
- Injection Linearity Test
- Carryover Test
Check the absorbance linearity of the instrument based on the height of the absorbance peaks detected upon injection of the aqueous caffeine solution. 2 For the high-pressure gradient system, set up 500 mL of water as solution A and aqueous caffeine solution (or acetone solution) as solution B.
Corrective Measures if Control Criteria are Not
10 Record the peak area value obtained for blank water in step 9 and five times the peak area value obtained for the 50 mg/L aqueous caffeine solution in step 7. 11 Calculate the proportion (%) of the blank sample's peak area value relative to the corresponding peak area for the 250 mg/L aqueous caffeine solution injected.
Resolving Common Troubleshooting Issues
Communication fails
Solvent Flow Does Not Stabilize
Solvent Delivery Pressure Increases
Baseline Does Not Stabilize
Noise Appears in Baseline
Poor Area Value Reproducibility
Carryover Does Not Disappear
Troubleshooting
Symptoms Presumably Due to an Error, Based on the
The amount of solvent in the mobile phase bottle is small and sucks in air. Due to incomplete solvent exchange, different solvents mix in the flow line and foam.
Symptoms Presumably Due to an Error, Based on a
System Controller
Pump
Autosampler
Column oven
UV Detector
System Installation Arrangement
System Tubing
Ultra High-Speed Analysis
Selecting a Mixer
Selecting the Cell
Removing Items Used to Secure the System during
Installing the Autosampler
Installing the Solvent Delivery Pump (LC-40D XR CL,
Installing the Detectors (SPD-40 CL)
Installing the Degassing Unit (DGU-405 CL)
Installing the Reservoir Tray
Installing the Column Oven (CTO-40C CL)
Installing Other Instruments
Cable Connections
Connecting the Power Cord
Connecting Optical Cables
Connecting a Degassing Unit (DGU-405 CL) to the
Tubing Connections
Installing Tubes and Joints for Draining Leakage
Connecting Low-Pressure Flow Channel Tubing
Connecting Tubing for High-Pressure Flow Channels
Checking Computer and System Controller
Setting the Computer IP Address
Specifying Computer Browser Settings
Specifying System Controller Settings
Other Installation Processes
Connecting to LabSolutions
Connecting the Computer to the System Controller
Installing Optional Column Management Functionality
5 Place the sample vials in the sample plate and place the plate on the sample rack. Contamination on walls within the flow cell makes it easier for air bubbles to collect in the cell (especially when water-based mobile phases are used).