Page 1 228-97194B Sep. 2022 Shimadzu Liquid Chromatograph Nexera CL series System Guide Read this manual thoroughly before you use the product. Keep this manual for future reference.
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Page 3 • The Safety Guideline specify important precautions for ensuring the system is operated safely. Be sure to read the Safety Guideline before using the product. • To ensure safe operation, contact your Shimadzu representative if product installation, adjustment, re-installation (after the product is moved), or repair is required.
Page 4: Introduction
• All rights are reserved, including those to reproduce this manual or parts thereof in any form without permission from Shimadzu Corporation. • Illustrations in this manual may differ from the actual product used. • Windows and Internet Explorer are registered trademarks or trademarks of Microsoft Corporation in the United States and/or other countries.
Page 5 Instruction Manuals Included with Respective Components These describe the accessories and options for each component, the name and function of various parts of the component, how to prepare, operate, and maintain the component, and so on. Nexera CL series...
Page 6 The following symbols are used in this manual: Indication Meaning Indicates an action that must not be performed. Indicates an action that must be performed. Indicates information provided to improve product performance. Indicates the location of related reference information. Nexera CL series...
Page 7: Operating Nexera Cl System
When, for example, analyzing samples that contain complex sample matrices such as serum, plasma or urine, it may be possible to obtain stabilized data by using an appropriate pre-treatment or an internal standard. Nexera CL series...
Page 8: Table Of Contents
Placing Samples ..............2.5.1 Preparing Sample Vials (or Sample Plates) ......2.5.2 Placing Samples in the Autosampler Performing a Single Run Analysis ....... 2.6.1 Check List to Confirm before Starting Analysis (while ........Waiting for Stabilization) Nexera CL series...
Page 9 Shutdown Function ................ 3.3.1 Executing the Shutdown Function 3.3.2 Executing a Batch File with a Shutdown Function ........... Specified in the File Auto-Diagnostics (Pulsation Check) and Auto-Recovery ................3.4.1 Auto-Diagnostics (Pulsation Check) ..........3.4.2 Auto-Recovery Nexera CL series...
Page 10 Inspection Item ........4.5.2 Testing Flowrate Accuracy Periodic Inspection of Autosamplers ..............4.6.1 Inspection Item 4.6.2 Injection Volume Accuracy Test (Total Injection Method ............Only) ......... 4.6.3 Temperature Accuracy Test Periodic Inspection of Plate Changers ....viii Nexera CL series...
Page 11 5.2.7 Carryover Does Not Disappear Troubleshooting ..........5.3.1 Symptoms Presumably Due to an Error, Based on the ..........Hardware Status 5.3.2 Symptoms Presumably Due to an Error, Based on a ..........Chromatogram List of Error Messages ........Nexera CL series...
Page 12 Connecting a Degassing Unit (DGU-405 CL) to the ........Solvent Delivery Pump Tubing Connections ........... 7.3.1 Installing Tubes and Joints for Draining Leakage .... 7.3.2 Connecting Low-Pressure Flow Channel Tubing 7.3.3 Connecting Tubing for High-Pressure Flow Channels ........(Connection Methods) Nexera CL series...
Page 13 Other Installation Processes ............... 7.5.1 Connecting to LabSolutions ..7.5.2 Connecting the Computer to the System Controller 7.5.3 Installing Optional Column Management Functionality ....(Column Management Device or “CMD”) Addendum Mobile Phase Properties ........Index ............Nexera CL series...
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Page 15: Overview
Overview Shimadzu Nexera CL series liquid chromatographs are designed to offer even higher levels of performance for meeting the increasingly diverse needs of liquid chromatograph users. System Features n A New Benchmark of Intelligence Mobile phase monitor Startup (FlowPilot) and shutdown at specified times Self-diagnostics (Pulsation Check) and auto-recovery　...
Page 16: Sil-40C Xr Cl Or Sil-40C X3 Cl Autosampler
1152 analytes (three MTP384 plates). Furthermore, up to three optional plate changers can be connected to load up to 44 microplates. In combination with temperature-control functionality, that enables continuous multianalyte analysis with high reliability. Nexera CL series...
Page 17: Cto-40C Cl Column Oven
600 mm 228-77025-06 Link Cable, 600 mm Link Cables 800 mm 228-77025-08 Link Cable, 800 mm 1000 mm 228-77025-10 Link Cable, 1000 mm 071-60821-08 For use in North America Power Cord 071-60845-01 For use in Europe Nexera CL series...
Page 18 The switch can be installed in front of the reservoir tray. The outlet has six sockets. • The switch can only be installed Power Outlet Unit, 6P on Nexera CL series or reservoir 228-65523-58 tray. Use the power outlet unit in combination with a Nexera CL series reservoir tray.
Page 19 The net is placed over the reservoir bottles Bottle Net Set for Earthquakes 228-54791-91 to prevent them from falling. This cable (approx. 1 m) extends the length Bottle holder extension cable 228-76080 of the bottle holder cable. Nexera CL series...
Page 20: Configuration List
Same as Kit A, except 170 mm wide for System Base Kit C 228-77036-43 plate changers For IVDR systems (for Europe),MPM-40 cannot be used. Configuration List Shimadzu Nexera CL series liquid chromatographs can be connected to the following units. Model Model Name High Performance Liquid 225-31400-55...
Page 21 LabSolutions Insight LCMS CL (P/N 225-45812-55) as multi-analyte quantitation software. LabSolutions LCMS or LabSolutions Insight software cannot be used. • In FDA compliant areas, LabSolutions LC/GC, LabSolutions LCMS, LabSolutions Insight, LabSolutions DB and LabSolutions CS can be used. Nexera CL series...
Page 22 Place samples in the autosampler. P.36 Perform a single run analysis. P.41 Perform a single run analysis. Perform postrun analysis. P.43 Perform realtime batch (sequential) analysis. P.48 Perform realtime batch Analyze multiple sets of data using Data (sequential) analysis. P.51 Browser. Nexera CL series...
Page 23 Refere Procedure Operation Purge columns with a column storage solution. P.55 Stop analysis and shut OFF the power supply. Shut down the instruments. P.56 Nexera CL series...
Page 24: Switching On The Power Supply And Opening The [Realtime Analysis] Window
If an optional PO-6P power outlet unit is used, then the main power can be switched ON/OFF from the front of the instruments. The switch is installed in the left front of the reservoir tray. Nexera CL series...
Page 25 Various keys are illuminated for directly specifying settings or Direct Key controlling the unit. *1 Different content is displayed for each unit. *2 Illumination may be different for some units. *3 Different content is displayed for each unit. Nexera CL series...
Page 26: Logging In To Labsolutions And Opening The [Realtime Analysis] Window
If logging in for the first time, select [Admin] in the [User ID] field and then click [OK]. For instructions on how to specify a user ID and password, refer to the LabSolutions Instruction Manual—System Users Guide. Nexera CL series...
Page 27 Displays icons for frequently used menu choices and analytical Toolbar instrument operations. Assistant Bar Displays icons for frequently used analysis operations. Status Displays the instrument status. Chromatogram Displays monitors for the detector signal, pressure values, and Monitor other measurement values. Nexera CL series...
Page 28 2 Analysis Name Description Control Panel Displays instrument parameter setting values and measurement (Instrument values. Parameter View) The layout and icons displayed can be customized. Nexera CL series...
Page 29: Specifying Analytical Conditions
This window is used to edit instrument parameters in the currently open method file. Analytical methods are edited in the instrument parameter editing window. Click [Data Processing Parameters] on the [Method] menu and specify analysis parameters before an analysis. Nexera CL series...
Page 30 Sets the end time of data acquisition. The data volume can be Detector Data reduced by specifying a shorter time than [LC Stop Time]. To Acquisition [End change the data acquisition start time, switch to the [Advanced] Time] Setting setting mode. Nexera CL series...
Page 31 % over 3 minutes, hold it there for 4 minutes, and then return it to the initial value, specify the time program shown. If the LC end time and pump mode settings are not specified correctly in the [Simple Settings] window, then the desired setting might be disabled in some cases. Nexera CL series...
Page 32 The [Init conc-replacement] setting is only displayed when the gradient pump mode is specified. The setting, if specified, purges flow channels from the specified pump to the autosampler with a mobile phase composition specified in initial gradient parameter settings. Nexera CL series...
Page 33 To only perform warm-up, select only the [Warm up] checkbox. If a column, mobile phases, and rinse solution are already installed, select the [Pump Purge], [Autosampler Purge], and [Warm up] checkboxes to perform the steps from auto-purging to warm-up at the same time. Nexera CL series...
Page 34 If the [Wait for Column Oven Temperature Stabilization] checkbox is not selected, then solvent is delivered at the flowrate specified in [Warm up Flow] for the time specified in [Warm up Time]. For precautions for auto-purging, refer to "2.3.2 Performing Auto-Purging". Nexera CL series...
Page 35 High-speed analysis in about 6 3.0 - 4.8 minutes High-speed analysis in about 12 4.8 - 7.2 3.333 minutes 7.2 - 13.0 13.0 - 26.0 1.428 26.0 - 36.0 1000 Over 12 minutes 36.0 - 45.0 2000 45.0 - 2000 Nexera CL series...
Page 36: Placing Mobile Phases And Autosampler Rinse Solutions
Insert suction filters into reservoir bottles containing the mobile phases and rinse solutions. Tag Displayed Port Name RINSE: R0 to R3 Autosampler rinse solutions SOLVENT:A Mobile phase A SOLVENT:B Mobile phase B SOLVENT:C Mobile phase C SOLVENT:D Mobile phase D Nexera CL series...
Page 37 In some cases, to prevent the formation of precipitates or emulsions, mobile phases must be purged with a separate solvent before purging the mobile phase remaining in the instrument or mixing it with another mobile phase. "2.9.3 Precautions When Replacing the Mobile Phase" P.64 Nexera CL series...
Page 38: Performing Auto-Purging
Click [AutoPurge] in the LabSolutions [Realtime Analysis] window. If the column is disconnected, clear the [Warm up] and [Init conc-replacement] checkboxes on the [AutoPurge] tab page in method settings. "2.2.1 Editing Methods" P.15 Nexera CL series...
Page 39 2.3 Placing Mobile Phases and Autosampler Rinse Solutions When a window appears to confirm executing auto-purging, click [OK]. Auto-purging is executed based on method settings. Nexera CL series...
Page 40: Manual Purging The Solvent Delivery Pump
Pull the solvent delivery pump drain tube out from the waste tank and insert a syringe, with a needle attached, into the end of the drain tube. Nexera CL series...
Page 41 When finished manually purging all flow channels, close drain valve (A) by turning it clockwise all the way until it stops. Close the right panel on the solvent delivery pump. Remove the syringe. Nexera CL series...
Page 42 • To manually purge , do not leave the column disconnected but connect the tubing with the coupling. Because the mobile phase flows into the analysis flow channel, liquid leaks in the column oven when the tubing is not connected, causing a leak error. Nexera CL series...
Page 43: Attaching The Column
Two rubber column clamps (B) for columns with an outer diameter from 6.3 to 9.5 mm are included as standard with the column oven. To clamp columns with an outer diameter from 9.5 to 12.7 mm, use an optional metal column clamp B8 (P/N: 228-15617-42). Nexera CL series...
Page 44 OFF cell temperature control and wait for the temperature to stabilize before resuming analysis (about 60 minutes). Switching the cell temperature control OFF can increase the baseline fluctuation range in response to room temperature fluctuations. Keep the room temperature constant to stabilize the baseline. Nexera CL series...
Page 45 Fully close the column oven door. To make sure the door does not open accidentally during analysis or open from contact with tubing inside the column oven, close the door until it makes an audible locking click sound. Nexera CL series...
Page 46: Warming Up The System
If auto-purging was already performed, clear the checkbox. User-specified parameter settings can be used for startup by editing the startup method. "2.2.1 Editing Methods" P.15 Click [Startup] on the toolbar in the LabSolutions [Realtime Analysis] window. Nexera CL series...
Page 47 Specify the date and time to begin startup, select the created method file, and select the [Perform LC Autopurge] checkbox. If the [Perform LC Autopurge] checkbox is selected, the auto-purge and warm-up actions specified in the selected method begin. If not selected, then [Pump] and [Oven] actions are started immediately. Nexera CL series...
Page 48: Placing Samples
In such cases, use the sample diluted or specify a lower sample suction speed setting value. Completely dissolve sample (A) by using a solvent composition that is equivalent to the mobile phase composition. Filter the sample through membrane filter (B). Nexera CL series...
Page 49 2.5 Placing Samples Fill the sample vial or well in a microplate or deep-well plate with the sample. Use genuine Shimadzu brand or Shimadzu-recommended sample vials and microplates. Using non-genuine or non-recommended vials or plates could cause flow channel blockage from septum debris or could prevent the needle from piercing the septum.
Page 50: Placing Samples In The Autosampler
After ten minutes, the temperature control mode is forcibly shut OFF. Plates for 1.5 mL sample vials are included as standard. To use other plates, be sure to specify the plate ID, specify the needle stroke distance, and adjust the plate position. Nexera CL series...
Page 51 　 Not doing so could cause part damage or equipment failure. Always pull out the sample rack by pulling horizontally with one hand and supporting the rack with the other hand. Nexera CL series...
Page 52 If the sample plate is shifted out of position, with respect to the sample rack, then the needle cannot be lowered properly into the specified vial or well, which could cause needle damage or other equipment failure. Nexera CL series...
Page 53 • If the main power switch is switched OFF on the back of the instrument during sample injection (while the status indicator is flashing yellow), the needle might stop in the lowered position in some cases. Pulling out the sample rack in that position could bend the needle. Nexera CL series...
Page 54: Performing A Single Run Analysis
(including tubing connections, for example) for any leaks. If a leak is discovered, tighten the leaking connection or Check for liquid replace the applicable consumables. Nexera CL series systems include leaks. a leak sensor, but it can take time before it responds to very small leaks, so also confirm that the pressure has reached the normally used level.
Page 55: Performing A Single Run Analysis
Performing a Single Run Analysis in the LabSolutions [Realtime Analysis] Window Specify analytical conditions. "2.2 Specifying Analytical Conditions" P.15 In the LabSolutions [Realtime Analysis] Window, click the (Single Run) icon. The [Single Run] window is displayed. Specify settings for a single run analysis. [Single Run] Window Nexera CL series...
Page 56 • If an emergency stop occurs during injection, the needle is moved to the drain port to discharge any suctioned sample remaining inside the needle and then the needle rinsing action starts. The analysis is finished when “Ready” is displayed in the main analysis window. Nexera CL series...
Page 57: Performing Postrun Analysis
Start LabSolutions and click [Postrun] in the main window. Double-click the [Postrun] icon. In LabSolutions DB and CS versions, a window for selecting the project is displayed. Select the project where the data to be analyzed was saved. Nexera CL series...
Page 58 • Data files can be listed by displaying Data Explorer and clicking the [Data] tab. • In LabSolutions DB and CS versions, the files can be filtered based on file name, instrument type, analysis date, or other criteria. Switch the Method View to the [Edit] mode and click the [Compound] tab. Nexera CL series...
Page 59 • Compound table setting parameters differ for quantitation methods other than [External Standard]. For more details about analysis parameters, refer to Help in LabSolutions or the LabSolutions Instruction Manual—Operators Guide. When finished editing method parameters, save the data by overwriting. Nexera CL series...
Page 60 By using the method settings specified here for the remainder of the analysis, data analysis results can be checked immediately after the analysis is finished. Nexera CL series...
Page 61: Displaying And Printing Data Reports
The data report is displayed in accordance with the previously specified report format settings. • To apply a different report format, drag and drop the report format file to the report. • Double-clicking an item in the report displays a [Property] window, where display settings can be specified. Nexera CL series...
Page 62: Performing Realtime Batch (Sequence) Analysis
Performing Realtime Batch Analysis in the LabSolutions [Realtime Analysis] Window Specify analytical conditions. "2.2 Specifying Analytical Conditions" P.15 Click [Quick Batch] in the LabSolutions [Realtime Analysis] window. The [Quick Batch] window is displayed. Specify the quick batch settings. Nexera CL series...
Page 63 This setting is linked to the Sample Type [Sample Type] setting on the left side of the [Batch Setting] window. Adds a new row to the batch table based on the specified Add Batch Table parameter settings. Nexera CL series...
Page 64 To add a batch row with no injection, set the [Vial Number] value to “-1”. Click [Start]. If the batch file is not saved, the [Save Batch File As] window is displayed. If the batch file is saved, the realtime batch analysis is started. Nexera CL series...
Page 65: Analyzing Multiple Sets Of Data Using Data Browser
Double-click the [Browser] icon. In LabSolutions DB and CS versions, a window for selecting the project is displayed. Select the project where the data to be analyzed was saved. Click [Quant Browser] on the assistant bar. Nexera CL series...
Page 66 [Table Style]. If method parameter settings need to be changed, switch the [Method View] to the [Edit] mode and then change the setting values. Parameter setting changes are applied to all data displayed in the [Quantitative Results View]. Nexera CL series...
Page 67: Outputting Summary Reports
Quantitation values for unknown samples are calculated based on the created calibration curve. 2.7.3 Outputting Summary Reports Check the [Quantitative Results View] content and then click [Summary Report] on the assistant bar. Nexera CL series...
Page 68 The data report is displayed in accordance with the previously specified report format settings. • To apply a different report format, drag and drop the report format file to the report. • Double-clicking an item in the report displays a [Property] window, where display settings can be specified. Nexera CL series...
Page 69: Stopping Analysis And Shutting Off The Power Supply
If a buffer solution was used as a mobile phase, rinse the flow channels with water after the analysis is finished. If the buffer solution dries, it forms crystals that could cause flow channel blockage, part wear, or flow cell contamination. Nexera CL series...
Page 70: Shutting Down The Instruments
ON. "3.3 Shutdown Function" P.75 To specify a shutdown method, create a method. Specifying a shutdown method enables the system to be shut down after cooling down, according to any specified parameters. "2.2 Specifying Analytical Conditions" P.15 Nexera CL series...
Page 71 2.8 Stopping Analysis and Shutting OFF the Power Supply Click [Shutdown] in the LabSolutions [Realtime Analysis] window. Specify [Shutdown] settings. "3.3 Shutdown Function" P.75 Click [OK]. The system is shut down according to the specified method. Nexera CL series...
Page 72: Setting Up Startup For Next Use
If an absorbance detector is used with a lower grade mobile Use HPLC grade or better phase than HPLC grade, mobile phase absorbance can overlap reagents for mobile with peaks at detected wavelengths, which can interfere with phases. quantitation of target components. Nexera CL series...
Page 73 Such bubbles can also solutions with mobile cause fluctuations or variability in retention times. phase. Nexera CL series...
Page 74 Nexera CL series...
Page 75 Replacing the Rinse Solution in the Solvent Delivery Pump Automatic Seal Rinsing Kit The automatic seal rinsing kit is used to automatically rinse the plunger seals and plungers in solvent delivery pumps. Select the rinse solution used for rinsing as described below. Nexera CL series...
Page 76 (B). Connect the coupling to the detector inlet tubing (A). Remove the cap (B) from the rinse bottle (A) in the automatic seal rinsing kit and fill the bottle with 100 to 200 mL of 10 % aqueous 2-propanol solution. Nexera CL series...
Page 77 Verify that rinse solution is discharged from the end of the FEP tube on the outlet end of the automatic seal rinsing kit. If rinse solution is discharged, stop pumping. Return the automatic seal rinsing kit to its previous configuration. Nexera CL series...
Page 78: Precautions For Selecting And Handling Mobile Phase Solvents
In addition if using the above solvents, rust on the stainless steel suction filter may contaminate the check valve and cause pressure fluctuations. Therefore, it may be necessary to change to a ceramic one, etc. Contact your Shimadzu representative for details.
Page 79: Precautions For Selecting An Autosampler Rinse Solution
Be sure to switch the degassing unit ON before rinsing. Leaving the degassing unit OFF during rinsing could cause air bubbles to accumulate in the rinse solution pipe, which can decrease sample injection accuracy. Select the autosampler rinse solution based on the mobile phase used, as follows. Nexera CL series...
Page 80 If using a solution containing more than 1 % formic or acetic acid or more than 0.1 % trifluoroacetic acid (TFA), for example, volatile components from the solution could corrode metal surfaces inside the instrument after long periods of continuous analysis, which could cause an instrument failure. Be careful of the following. Nexera CL series...
Page 81 [Move Autosampler needle to Z Home] checkbox in the shutdown settings. Using this function can minimize the impact of corrosion by having the Z-mount wait at a location away from the rinse port during shutdown. "2.8.3 Shutting Down the Instruments" P.56 Nexera CL series...
Page 82: Using Buffer Solutions As A Mobile Phase
5 and 6. Depending on the pumping pressure, rinse solution can mix with mobile phase to form salt precipitates. Nexera CL series...
Page 83 Replacing the tube with the LPGE-OUT tube will increase the system delay volume, so be sure to change the delay correction volume setting accordingly. For more information, refer to the solvent delivery pump instruction manual "3.5.4 [SYSTEM] Setting Group, Inputting the tube volume". Nexera CL series...
Page 84: Application Operations
-purge the autosampler, and warm up the system (from starting solvent pumping to column equilibration). Create a method to be specified for the startup process. "2.2 Specifying Analytical Conditions" P.15 The following describes an example of method settings required for the startup function. Nexera CL series...
Page 85 Specify the method file created in the [Startup] setting window, select the [Perform LC Autopurge] checkbox, and click [OK]. That schedules the startup process. Because method files currently being used in the [Data Acquisition] window cannot be specified, specify any other method than the one currently in use. Nexera CL series...
Page 86: Executing A Startup Process And Starting A Batch Analysis At A Specified Date/Time
"3.2.1 Executing the Startup Process" P.70 Open and edit the batch file. Open the batch file to be executed after the startup process. Open the [Settings] tab page. In the LabSolutions [Realtime Batch] window, click [Settings]. Batch settings are displayed. Nexera CL series...
Page 87 Specifies using the method file specified in the first the first row row of the batch file for executing the startup process. If selected, the auto-purging setting specified in the Perform LC Autopurge method file is executed during startup. Nexera CL series...
Page 88 When batch analysis starts based on a batch file that specifies a startup process, the batch analysis is registered in the batch queue. The registered batch analysis first executes the startup process at the date/time specified by the startup function. Nexera CL series...
Page 89: Shutdown Function
• If the column is to be rinsed, then a method is specified that rinses the column according to the rinsing method specified in the column instruction manual. Display the shutdown settings. Click [Shutdown] in the LabSolutions [Realtime Analysis] window. Shutdown settings are displayed. Nexera CL series...
Page 90 ・ If the [Power Off after shutdown] checkbox is Shutdown Settings cleared, the system switches to the sleep mode. Settings can be specified for switching OFF actions by individual instruments. For more information about specifying settings for individual instruments, refer to Help in LabSolutions. Nexera CL series...
Page 91: Executing A Batch File With A Shutdown Function Specified In The File
To cancel a shutdown function while a batch analysis is being executed, the batch analysis must be stopped. To specify the shutdown function in a shutdown method, create the method. "3.3.1 Executing the Shutdown Function" P.75 Nexera CL series...
Page 92 3 Application Operations Open the [Settings] tab page. In the LabSolutions [Realtime Batch] window, click [Settings]. Batch settings are displayed. Nexera CL series...
Page 93 Sets whether or not to perform the cool-down process Shutdown Method File using the specified method file when the shutdown process is executed. Specifies how long to operate the system before Cool down Time executing the shutdown process. Nexera CL series...
Page 94 For more information about specifying settings for individual instruments, refer to Help in LabSolutions. Click [OK]. Shutdown settings are saved in the batch file. Start the batch analysis. The shutdown process is executed after the batch analysis is finished. Nexera CL series...
Page 95: Auto-Diagnostics (Pulsation Check) And Auto-Recovery
Recovery from Pressure Fluctuations The Auto-Recovery function is used to recover from solvent delivery problems resulting from air bubble ingress. • This function is only available when using LabSolutions. • This function is only enabled for batch analysis using LabSolutions. Nexera CL series...
Page 96 3 Application Operations Make sure the solvent delivery pump Auto-Diagnostics (Pulsation Check) function is ON. The [Auto-Diagnostics] setting (using the Auto-Diagnostics function) must be “1” in the [CALIBRATION] information group. Nexera CL series...
Page 97 LabSolutions [Realtime Analysis] program. Click the (Settings) icon on the [Realtime Batch] assistant bar. Click the [Auto Recovery] tab and specify Auto-Recovery settings. Then click [OK]. For more details about the parameter settings, refer to Help for LabSolutions. Nexera CL series...
Page 98 LabSolutions will stop the batch analysis. After stopping the batch analysis, solvent delivery and column temperature control continue. Click the (Start Realtime Batch) icon on the [Realtime Batch] assistant bar to start the batch analysis. Nexera CL series...
Page 99: Autosampler Functions
Reagents in vials can be added to vials for analysis. Up to three types of reagents can be added and a mixing action can be included after adding reagents, so that analysis can be performed by adding reagents with no concentration non-uniformities. Nexera CL series...
Page 100 Stack Injection Samples can be detected based on multiple peaks in the same data file by successively injecting the same sample. Nexera CL series...
Page 101 3.5 Autosampler Functions n Overlap If the same analysis is performed successively multiple times, then analysis cycle time can be shortened by executing the next pretreatment action before the previous analysis is finished. Nexera CL series...
Page 102: Cut Off Loop Function
Solvent delivery pump High-pressure valve Sample loop Needle Injection port Injection port tube Flow channel to column Nexera CL series...
Page 103: Low Dispersion Injection
Batch action functions in batch tables can be used to select actions to execute if an analysis fails a system check or system suitability test. This section describes an example of operations used to cancel a realtime batch analysis when an analysis fails the system suitability test. Nexera CL series...
Page 104 Click the [Action] cell and then select [Stop (Wait Next Queue)] in the list. The [Test] cell can also be used to determine system suitability results based on QA/QC calculation or system check results. For information about QA/QC calculations, refer to Help. Nexera CL series...
Page 105 Returns to the row where [Goto] was previously Return executed. Click (Save Batch File) on the toolbar. The settings are saved to batch file. For information about operations used to create batch tables, refer to the LabSolutions Instruction Manual—Operators Guide. Nexera CL series...
Page 106 If an analysis fails the test with settings configured that way, the last row of the batch table is executed after any row with an [Action] setting specified. That means the instrument can be shut down according to the instrument parameters specified in the method file specified in the last row. Nexera CL series...
Page 107: Using Acto (Analytical Condition Transfer And Optimization) Functions
The setting the internal volume of the system controller by the field engineer. (Contact your Shimadzu representative.) • LC-40 CL Series Solvent Delivery Unit Instruction Manual "3.5.4 [SYSTEM] Setting Group/Inputting the tube volume《INNER VOLUME》"...
Page 108 [After in [Adjustment volume]. Injection] This setting is used for migrating from a system with a large delay volume to a system with a small delay volume. Nexera CL series...
Page 109: Method Migration Functions
In LabSolutions [Realtime Analysis] window, set the pumping mode to the target gradient mode. In LabSolutions [Realtime Analysis] window, click [File]-[Method Transfer] and select the method file to migrate (the method file used for the previous system). Nexera CL series...
Page 110 • To adjust for differences in the gradient delay volumes of the respective systems, select the [Adjust the gradient start] checkbox. • To use a time program to adjust the concentration gradient delay time, select the [Gradient program] checkbox in [How to adjust]. Nexera CL series...
Page 111 • Because the [P.Max of Pump] value is not applied either, also specify [Maximum] on the [Pump] tab page, based on the given column and flowrate. If the [Gradient start adjustment] setting is used, make fine adjustments to the delay volume, as necessary. Nexera CL series...
Page 112: Useful Data Analysis Functions
Parameter settings must be adjusted for baselines with large undulations that exceed the automatic baseline estimation range, as shown below. In the [Method View], click [Edit]. Change the [Detection Threshold] setting from the default value of 5 to 100. Nexera CL series...
Page 113 3.8 Useful Data Analysis Functions In the [Method View], click [View]. Nexera CL series...
Page 114: Column Management
• The column manager is not available for LabSolutions LC/GC and LabSolutions LCMS. Available in LabSolutions DB and LabSolutions CS. • The column manager is not available for IVDR System. • For details on CMD, refer to the instruction manual (1.3 Optional Parts) of the oven (CTO-40C CL). Nexera CL series...
Page 115: Column Management Workflow
"Configuring the column (from the column manager)" "Analysis" "Viewing the acquisition log in the column manager" "Viewing the deterioration status of columns in "3.9.5 Viewing the Acquisition Log" the column browser" "Viewing chromatograms in the column browser" Nexera CL series...
Page 116: Setup For Labsolutions
In the [Main] assistant bar, click the [System Configuration] icon. In [Available Modules], select [Column] and then click the [→] button before clicking the [OK] button. After a link beep sounds, the [Column] information appears in the [Column Oven] tab. Nexera CL series...
Page 117 "Installing your CMD and column to the instrument" P.108 Start the [Realtime Analysis] program of LabSolutions. "2.1.2 Logging in to LabSolutions and Opening the [Realtime Analysis] Window" P.12 In the LabSolutions tool bar, click [CMD Information]. The [CMD Information] window appears. Nexera CL series...
Page 118 3 Application Operations After the message "A CMD specified in a previous version was inserted. Set the column information again after clearing the CMD information." appears, click the [OK] button. Click the [Clear CMD Info.] button. Nexera CL series...
Page 119 The initialization of the CMD is complete when the display of [CMD 1 (Oven)] changes from [Previous Version and CMD-Instrument Inconsistency] to [Not registered]. According to "Registering the column in the CMD" P.109, register a new column in the CMD. Nexera CL series...
Page 120: Column Registration And Replacement (Discard)
Starting the column manager and registering a column In the [Administration Tools] window, double-click the (Column Manager) icon. This starts the column manager. Select [LC] and then, from the [Add] menu, click [New]. Nexera CL series...
Page 121 [Add]-[Copy] button, change [Column ID], and then edit necessary items before registering the column. • Perform this procedure before installing the column to the instrument. • Exit the column manager as needed. (It is not always necessary to exit the column manager.) Nexera CL series...
Page 122 Connect the CMD cable to the "CMD" connector of CTO-40C CL. Fix the CMD to the column with a cable tie coming with the CMD. Install the column equipped with the CMD to the column oven. Connect the CMD to the CMD cable. Nexera CL series...
Page 123 For details on configuration of each unit, refer to the instruction manual of each unit. Start the [Realtime Analysis] program of LabSolutions. "2.1.2 Logging in to LabSolutions and Opening the [Realtime Analysis] Window" P.12 In the LabSolutions tool bar, click [CMD Information]. The [CMD Information] window appears. Nexera CL series...
Page 124 • For details on the setting items, refer to the help of [CMD Information]. • Be sure to enter the name and column ID. • Some setting items cannot be registered in CMD. Use the column manager to set the items that cannot be registered. Nexera CL series...
Page 125 After the following message appears, click the [OK] button. After registration in CMD, the LC status changes to [CMD-Method Column Mismatch]. In the [Column Oven] tab, click the [Read from CMD] button. Click the [Close] button. The [CMD Information] window closes. Nexera CL series...
Page 126 This section describes how to replace a column together with the CMD after the separation capability deteriorates. Connect the CMD of the column to be discarded to the CMD cable. In the LabSolutions tool bar, click [CMD Information]. The [CMD Information] window appears. Click the [Discard Column] button. Nexera CL series...
Page 127 If the new column has the same specifications as the old one, after selecting the old column, click [Add]-[Copy], which saves you the trouble of data entry. Be sure to change the column ID at this time. Nexera CL series...
Page 128 If the new column has the same specifications as the old one, after selecting the old column, click [Add]-[Copy], which saves you the trouble of data entry. Be sure to change the column ID at this time. Nexera CL series...
Page 129: Analysis With Labsolutions
After a link beep sounds, the column information is reflected and the [CMD-Method Column Mismatch] status changes to one such as [Ready]. Without the mobile phase purged, the status may change to another such as [Pump A Wait Purge]. Nexera CL series...
Page 130 Configuring the column (from the column manager) This section describes how to configure the column without using the CMD. In the [Column Oven] tab, click the [Settings] button. Click the [Refer to Column List] button. The [Column List] window appears. Nexera CL series...
Page 131 Select the column to be used in analysis and then click the [Select] button. The information on the selected column is reflected. Click the [OK] button. After a link beep sounds, information on the selected column is reflected in [Column]. Nexera CL series...
Page 132: Viewing The Acquisition Log
Enables you to specify filtering conditions for analysis results. Displays filtered analysis results. For display contents, refer to the help of the column manager. You can print the results. For details, refer to the "Column Manager Operating Manual" provided with LabSolutions. Nexera CL series...
Page 133 With the history of column analysis results displayed, click the [Column Data Browser] menu. Click the compound name of the component peak you wish to check. This requires that the compound table be already edited (compounds be already identified) in analysis data. Results are displayed for identified data. Nexera CL series...
Page 134 3 Application Operations Click the column parameter you wish to view. Injection time series of change in the parameter appears in the lower section of the window. Nexera CL series...
Page 135 3.9 Column Management n Viewing chromatograms in the column browser Click [Load Chromatogram]. The browser displays the chromatograms of the data listed in the table by overlaying them. Nexera CL series...
Page 136: Hardware Validation
Hardware validation is broadly classified as component validation or overall system validation, but both types of validation are required. Each Shimadzu product is shipped with inspection results obtained when it was manufactured. However, this chapter describes, separately for individual components and the overall system in case the instrument performance must be validated, the inspection procedure and guidelines for inspection pass/fail criteria.
Page 137: Routine Inspections
Used to verify the accuracy of the solvent delivery pump flowrate. Stopwatch The stopwatch specifications must guarantee performance within 5'30" ±0.3 seconds. Used to verify the accuracy of the solvent delivery pump flowrate. Volumetric Flask Have a 5 mL volumetric flask available. Nexera CL series...
Page 138 Thermometer/temperature gauge and recorder specifications must guarantee a rated accuracy of ±1.0 ℃ within the temperature range (0 to 100 ℃) required for inspecting equipment. n Standard Reagents for Performance Inspections Purchase the standard reagents from Shimadzu. Reagent Description Aqueous Caffeine...
Page 139 After obtaining the correction value, calculate the measured temperature using the following formula. [Measured temperature] = [Thermometer reading] + [Set temperature] Nexera CL series...
Page 140 *1 Since the SPD-40 CL wavelength accuracy test is performed in the cell enclosure, the 20 mg/L sample may have a high absorbance value and may not be tested accurately. Therefore, do not use this sample. *2 This item is used with ferrules and male nuts. Nexera CL series...
Page 141: Periodic Inspection Of Solvent Delivery Pumps
Display [COMP] in [PARAMETER] group. Record the current setting value and reset the value to 0.45 Display [FLOW CHECK] in [VALIDATION] group and press the [Enter] key. Specify the flowrate accuracy passing criterion value (%). Use the keypad to enter the setting value and then press [Enter]. Nexera CL series...
Page 142 When prepared for measurement, press [Enter] again and measure the time required for 5 mL to collect in the volumetric flask. Enter the time measurement value (seconds). The flowrate accuracy (%) pass/fail result is displayed. Control Within ±2.5% Criteria Nexera CL series...
Page 143 The flowrate accuracy (%) pass/fail result is displayed Control Within ±3 % Criteria When the test is finished, repeat steps 3 to restore the [COMP] setting value to the value recorded in step 3. Nexera CL series...
Page 144: Periodic Inspection Of Autosamplers
Specify the following analytical condition settings. ANALYSIS TIME 0.1 min RINSE TYPE 0　(no rinsing) ML PURGE VOL 100 µL RINSE SPEED 35 µL/s 5 µL/s (50 µL loop) SAMPLE SPEED 15 µL/s (100 µL loop) AIR GAP Nexera CL series...
Page 145: Temperature Accuracy Test
50 µL injections: Injection accuracy is within ±2.0 % Criteria 5 µL injections: Injection accuracy is within ±5.0 % If control criteria are not satisfied, refer to contact your Shimadzu representative. 4.6.3 Temperature Accuracy Test n Scope of Inspection Sample cooler temperature accuracy is calculated using external temperature sensor measurement results.
Page 146 Place the 1.5 mL sample vial plate at the front of the sample rack (position 1) and then insert the sample rack. Set the sample cooler temperature setting to 4 ℃ and start temperature control. Nexera CL series...
Page 147 Given conditions that do not exceed a maximum 30 ℃ room temperature or 60 % humidity Control Criteria Temperature Using 1.5 mL sample vial plates: 4 ℃ ±3.0 ℃ setting accuracy Nexera CL series...
Page 148: Periodic Inspection Of Plate Changers
When the plate is set and the door is closed, the arm lifts the plate and pulls it into the instrument to identify the plate type and adjust its position. Once the operation is completed, the status indicator on the plate stack illuminates green. Nexera CL series...
Page 149 After that, confirm that the status indicator on the plate stack illuminates green, and then perform the procedure in this step. Set the sample cooler temperature setting to 4 ℃ by the operation panel of Autosampler and start temperature control. Nexera CL series...
Page 150 Given conditions that do not exceed a maximum 30 ℃ room temperature or 60 % humidity Control Criteria Temperature Using 1.5 mL sample vial plates: 4 ℃ ±3.0 ℃ setting accuracy Nexera CL series...
Page 151: Periodic Inspection Of Column Ovens
Connecting Parts for Inspection Calibrate the temperature gauge thermocouple used for measurements and measure the correction value. It does not need to be recalibrated for each measurement, but calibrate it regularly according to a method specified separately. Nexera CL series...
Page 152 Then press the [OVEN] key. Temperature adjustment starts. Wait until [READY] is displayed on the screen and then leave the system undisturbed for ten minutes. On the initial screen, press the [→] (right arrow) key one time. Nexera CL series...
Page 153 Continue by pressing the [Enter] key. Enter the temperature measured using a digital temperature gauge and check the temperature accuracy. Temperature accuracy is within ±2.0 ℃ and Control Criteria [ACCURACY GOOD] is displayed If the accuracy fails test, contact your Shimadzu representative. Nexera CL series...
Page 154: Periodic Inspection Of Uv Absorbance Detectors
205 nm, 245 nm, and 273 nm. The instrument automatically determines whether the absorption peaks for caffeine are within the 205.0 nm ± 2 nm, 245.0 nm ± 2 nm, and 273.0 nm ± 2 nm ranges, respectively. Nexera CL series...
Page 155 Use a syringe to fill the flow cell with caffeine and then press [Enter]. The window on the right is displayed for about 30 seconds. Then the caffeine is measured. When the caffeine measurement is finished, the screens shown on above are alternately displayed. Next, the background is measured. Nexera CL series...
Page 156 Inspection Procedure (2) Inspecting with Manual Wavelength Change. Using this method, wavelength can also be checked based on theophylline. For more details, contact your Shimadzu representative. Scope of Inspection Aqueous caffeine solution is used to manually check wavelength accuracy at 205 nm, 245 nm, and 273 nm.
Page 157 • If the deviation is more than ±2 nm, then follow the instructions in " Corrective Measures if Wavelength Accuracy Does Not Satisfy Control Criteria". • After inspection, be sure to reset the [Z WAVE] setting to "0". Nexera CL series...
Page 158 Inspection Procedure (3) Inspecting with Wavelength Scan Function (Using the LabSolutions) Using this method, wavelength can also be checked based on theophylline. For more details, contact your Shimadzu representative. Scope of Inspection Aqueous caffeine solution is used to check wavelength accuracy at 205 nm, 245 nm, and 273 nm with wavelength scan function .
Page 159 SPD-40 CL Instruction Manual Calibrate wavelengths. Wavelength calibration was not 4.8 Performing Wavelength Calibration in performed properly. the SPD-40 CL Instruction Manual Optical system problem (degradation of Perform optical system maintenance. (Contact a mirrors, window, etc.). Shimadzu representative. ) Nexera CL series...
Page 160: Periodic Inspections Of Systems
2-Propanol and 10 min 2 mL/min solvent as mobile phase then water each Used system with water/organic solvent mixture or water and water-compatible organic solvent Water 2 mL/min 10 min (such as methanol or acetonitrile) as mobile phase Nexera CL series...
Page 161: Inspection Item
Contents in Set of 5 Aqueous Caffeine Solution Concentrations Name Solution 1 Solution 2 Solution 3 Solution 4 Solution 5 Set of 5 aqueous caffeine 20 mg/L 30 mg/L 40 mg/L 50 mg/L 60 mg/L solution concentrations Nexera CL series...
Page 162 If the absorbance for solution 5 does not reach 1.0 AU, remove the 2 m × 0.5 mm I.D. flow restrictor and only use the 4 m × 0.1 mm I.D. flow restrictor. After the baseline stabilizes, successively inject the samples (solutions 1 to 5). Nexera CL series...
Page 163: Noise/Drift Test
CL Instruction Manual Mirror or window plate Recalibrate linearity and perform optical system degradation has occurred. maintenance. (Contact a Shimadzu representative.) 4.10.3 Noise/Drift Test n Scope of Inspection Checks that noise and drift values satisfy specified criteria. n Parts Required for Inspection...
Page 164 Determine noise as the average of noise bands for all 30 intervals. (Refer to: ASTM 　E1657-98) Drift Evaluation Method The method of calculating the drift value differs between automatic and manual calculations. • In the case of automatic calculation Nexera CL series...
Page 165 4, and this is taken as the amount of change over 1 hour, which is taken as drift (AU/h or RIU/h). Determine drift (AU/h) as the fluctuation within an hour of data, calculated as four times the baseline fluctuation for the 15-minute period. Nexera CL series...
Page 166: Gradient Concentration Test (Gradient System)
4 m × 0.1 mm I.D. and 2 m × 0.5 mm I.D. Flow Restrictor Purified Water HPLC grade or equivalent Preparation method (for Caffeine) • Using Shimadzu standard reagents for performance inspection Measure 20 mL of caffeine (250 mg/L, P/N: Aqueous solution of 228-45725-06) and add water to make 500 Caffeine...
Page 167 0 % (B.Conc = 0), 10 % (B.Conc = 10), 50 % (B.Conc = 50), 90 % (B.Conc = 90), 100 % (B.Conc = 100) Measure the signal level when the baseline stabilizes. Nexera CL series...
Page 168 Prepare and execute the following gradient program. High-pressure Gradient Program TIME FUNC VALUE 0.00 B.Conc 0.01 B.Conc 100.0 10.00 B.Conc 100.0 10.01 B.Conc 90.0 15.00 B.Conc 90.0 15.01 B.Conc 50.0 20.00 B.Conc 50.0 20.01 B.Conc 10.0 25.00 B.Conc 10.0 25.01 B.Conc 30.50 Stop Nexera CL series...
Page 169 15.00 D.Conc 90.0 15.00 C.Conc 10.0 15.01 D.Conc 50.0 15.01 C.Conc 50.0 20.00 D.Conc 50.0 20.00 C.Conc 50.0 20.01 D.Conc 10.0 20.01 C.Conc 90.0 25.00 D.Conc 10.0 25.00 C.Conc 90.0 25.01 D.Conc 25.01 C.Conc 100.0 30.50 Stop Nexera CL series...
Page 170: Reproducibility Test
*1 The flow restrictor is removed for system configurations that result in abnormal pump pressures at the specified flowrate. *2 Be sure to use a degassing unit to degas each mobile phase. Connect the flow restrictor between the injector outlet tube and the detector. Rinse the flow channels with an appropriate solvent. Nexera CL series...
Page 171 Confirm that liquid is flowing from the detector outlet tube and no liquid is leaking from any connections. Monitor the baseline. Once the baseline is stable, adjust the detector zero-point. Inject 10 µL of water and confirm that no peaks are observed. Nexera CL series...
Page 172 : Number of analyses : Retention time (or peak area) of individual peaks : Mean value of X1...Xn : Standard deviation : Coefficient of variation Peak area CV %　is within　1.0 Control Criteria Retention time CV %　is within　 0.5 % Nexera CL series...
Page 173: Injection Linearity Test
Calculate the correlation coefficient contribution rate (R ) based on peak area results. Control Criteria 0.9990 or higher 4.10.7 Carryover Test n Scope of Inspection This validation inspects the carryover based on the measured peak area ratio. Nexera CL series...
Page 174 Add 1 mL of water in one other sample vial and seal it with a septum. "2.5 Placing Samples" Place the sample vials in the sample plate and place the plate on the sample rack. Inject 10 µL of blank water. Inject 10 µL of 50 mg/L aqueous caffeine solution. Nexera CL series...
Page 175: Corrective Measures If Control Criteria Are Not Satisfied
However, refer to the corresponding instruction manual for each instrument for instructions on how to implement the troubleshooting. If not sure what the cause is, contact your Shimadzu representative. Also, if the cause of not satisfying control criteria is unknown or if unsure about the troubleshooting or maintenance process, contact your Shimadzu representative.
Page 176: Troubleshooting
If an Instrument or Workstation Software Error Message is Displayed "5.4 List of Error Messages" P.194 In LabSolutions, error messages can be viewed in the LabSolutions [Output Window]. n Workstation Operating Problems Refer to Help in the workstation software. Nexera CL series...
Page 177: Resolving Common Troubleshooting Issues
ON/OFF? ÈYES Are workstation configuration settings Reconfigure the consistent with the actual workstation configuration Æ instrument configuration in settings. the system? ÈYES Contact your Shimadzu representative. "7.4 Checking Computer and System Controller Connections" P.305 Nexera CL series...
Page 178: Solvent Flow Does Not Stabilize
Is there any leakage around Rinse/replace the check Æ the pump head? valve. ÈYES Replace the plunger seal. Check the plunger status. If any damage or contamination is discovered, replace the plunger and diaphragm at the same time. Nexera CL series...
Page 179: Solvent Delivery Pressure Increases
ÈYES ÈYES Inspect the mixer, solvent Replace the high pressure delivery pump, and mobile valve rotor and then phase bottles. inspect/replace the stator. Nexera CL series...
Page 180 Debris from the debris, it could potentially cause tubing blockage. If needle Needle Seal Installed misalignment at the injection port is observed, the injection port in the Injection Port position must be adjusted. SIL-40 CL Series Instruction Manual Nexera CL series...
Page 181 In such cases, the debris that would cause the tubing blockage can Valve Rotor/Stator cause blockage at the column inlet. Therefore, first try the countermeasures indicated above. These same precautions also apply when this unit is used in combination with a non-Shimadzu LC system. SIL-40 CL Series Instruction Manual Required...
Page 182: Baseline Does Not Stabilize
・ Change in the dissolved oxygen column? content in the mobile phase. Keep the mobile phase temperature constant and degas it thoroughly. ・ Change in mobile phase composition. Seal reservoir bottles and leave them in a location with minimal temperature variations. ÈYES Nexera CL series...
Page 183: Noise Appears In Baseline
While pumping solvent, apply pressure inside the cell by pressing a fingertip, a rubber object, or other item against the end of the cell outlet tube. Repeat that blocking and releasing cycle several times to stretch and compress the bubbles until they flow out. Nexera CL series...
Page 184 (The slot in cell window fastening screws is 1 mm wide and 8 mm long. The regular screwdriver part number is S086-10504-18.)　If tightening the cell window fastening screws does not resolve the leak, contact a Shimadzu representative. Nexera CL series...
Page 185: Poor Area Value Reproducibility
ÈYES Does reproducibility vary depending For some vial materials, samples on the component or do values could adsorb to vial walls. Change Æ decrease when left in vials for a the vial material. while? Nexera CL series...
Page 186 (b) of the tray (B) by the hook (a) at the bottom. Pushing in the switch releases the hook from the tray and enables you to pull up the switch. • The figure is the side view of the instrument seen from the left. Nexera CL series...
Page 187 Stop the purge operation and attach the piping and soft switch as it was. After the manual purge is complete, purge the Autosampler as usual. Nexera CL series...
Page 188: Carryover Does Not Disappear
In the cases of LC and other high-sensitivity analysis, carryover can also occur from sources to which samples are exposed, other than flow channels, such as mobile phase bottle contamination. Nexera CL series...
Page 189: Troubleshooting
(The solvent flowrate display Specify the flowrate. pump delivery pump remain at “0”? does not Is an error function.) displayed (such as Solvent delivery Click [CE] and eliminate the cause “P-MAX” or pump of the error. “P-MIN”)? Nexera CL series...
Page 190 Is the check valve Solvent delivery first. Then rinse it by pumping defective? pump 2-propanol. If rinsing the valve is not effective, clean the check valve ultrasonically or replace it. Nexera CL series...
Page 191 Replace the plunger seal. If the Is there leakage Solvent delivery plunger seal service life was from the plunger pump extremely short, then also replace seal? the plunger. Is the drain valve Solvent delivery Close the drain valve. open? pump Nexera CL series...
Page 192 Autosampler the high stator. stator decreased? pressure valve. There is leakage Is the airtightness from a gap in Inspect/replace the rotor and of the rotor and Autosampler the low pressure stator. stator decreased? valve. Nexera CL series...
Page 193 Specify an appropriate cell temperature-control temperature. The cell temperature-control Cell setting range varies depending on Is the Temperature the room temperature, column temperature Control Failure Detector temperature, solvent delivery setting pump flowrate, and cooling appropriate? tubing. SPD-40 CL Instruction Manual Nexera CL series...
Page 194 Detector SPD-40 CL Instruction alignment performed Manual properly? Is [Autosampler Control] set to The injection [Offline] in Set the [Autosampler Control] action does not Autosampler autosampler setting value to [Online]. occur. [System Setting] settings? Nexera CL series...
Page 195 ・ The flow channel selection valve for the column oven is not displayed. Make sure the [System P.Max] The [System P.Max] A "P.Max" system error setting in system controller setting value is not occurs. configuration settings is reasonable. appropriate. Nexera CL series...
Page 196 Power supply which can sometimes result in injection port. power switched leaking. OFF? Do not switch OFF or shut down the autosampler when the pump is pumping solvent. Nexera CL series...
Page 197 In the event that the [Error] lamp lights up or any liquid continuously flows out even after the restarting is repeated several times, the degassing unit may have failed. Contact your Shimadzu representative. Nexera CL series...
Page 198: Symptoms Presumably Due To An Error, Based On A Chromatogram
If rinsing is not effective, then replace the needle or tubing. Is the needle seal If the needle seal is worn, replace Autosampler worn? Were flow channels rinsed Autosampler Click [PURGE] or [RINSE]. inadequately? Nexera CL series...
Page 199 Autosampler volume is discovered, properly flow channel reconnect the connection. connections? Tighten the connection. If the leak Is there leakage persists after tightening the from a flow Entire system connection, then replace the channel? connection. Nexera CL series...
Page 200 Change the needle stroke to a Autosampler that the hole is smaller or larger value. covered, or not reach the sample solution? Confirm whether the detector is Is the detector functioning properly and functioning Detector implement corrective actions properly? accordingly. Nexera CL series...
Page 201 (solvent delivery contaminated, Autosampler pressure: 1 to 5 MPa). such as the 2 Flush with purified water for needle or sample about 30 minutes at a flowrate loop? of 9 mL/min (solvent delivery pressure: 1 to 5 MPa). Nexera CL series...
Page 202 Check the accuracy of the column times for target oven temperature. If not components are Has the column appropriate, adjust the significantly temperature Column oven temperature accuracy. different from deviated? when measured CTO-40 CL Instruction using a different Manual system. Nexera CL series...
Page 203 If it phase and Mobile phase decreases, the mobile phase might column in stable and column contain impurities or the column condition? might not be fully equilibrated. Check the mobile phase, column, analytical conditions, and other factors. Nexera CL series...
Page 204 Detector cell. cell? The baseline shifts. SPD-40 CL Instruction Manual Has the connection where the lamp or flow If there is looseness or a gap, Detector cell is attached reattach it properly. loosened or developed a gap? Nexera CL series...
Page 205 Click the [ZERO] button in the value dropped to [RUN] window. Adjust the zero the maximum point. UV detector negative value Also, set [ZERO] to [Enter]. Controller (or increased to SPD-40 CL Instruction the maximum Manual positive value)? Nexera CL series...
Page 206 Alternatively, visually check for Are there irregular any bubbles flowing in the flow bubbles intervals. Detector cell. accumulated in If there are bubbles, disassemble the flow cell? and clean the flow cell. SPD-40 CL Instruction Manual Nexera CL series...
Page 207 *2 : Inspect for suction filter blockage: Remove the filter and record the pressure waveform. If the pressure waveform improves when disconnected, that suction filter is clogged. *3 : An “air cell” refers to a flow cell with all cell lenses and cell gaskets removed and not containing mobile phase. Nexera CL series...
Page 208: List Of Error Messages
An error occurred in 0x2F18 (LAN cable, HUB etc). workstation. Check the communication with 0x2F20 If the error persists, restart communication the workstation. 0x2F21 both the system controller and environment. 0x2F22 the workstation. 0x2F23 0x2F24 0x2F25 0x2F26 0x2F27 0x2F28 Nexera CL series...
Page 209 0x4001 etc.). workstation. Check the communication with 0x4004 If that does not resolve the communication the workstation error, contact a Shimadzu sales environment. could not be or service representative. established. Check the instrument There was no connection status and perform...
Page 210 If an error occurs each time settings were The backup parameter of the system controller is reinitialized the system controller has started, contact a Shimadzu (cleared). been damaged. The sales or service representative. 0x0006 method file and sequence file were changed. Check on the method file and sequence file.
Page 211 0x5D10 Measured value was not the mobile phase If the error still occurs, unplug correctly obtained. monitor. the power cord for the main unit and contact a Shimadzu service or sales representative. Nexera CL series...
Page 212 If the error still occurs, unplug sensor is connected Check that the the power cord for the main correctly. sensors to be used unit and contact a Shimadzu are properly service or sales representative. connected. Check that the bottle holder is properly connected and then...
Page 213: Pump
Controller Screens FATAL Communication Switch the instrument power CONNECTION A-B failed between 0x5000 OFF and contact a Shimadzu pumps A and B in Pump: System error has sales or service representative. the main system. occurred. WARNING The system can continue to be...
Page 214 If it occurs frequently, switch The instrument was 0x5F05 reasons, because a the instrument power OFF and restarted for safety system error contact a Shimadzu sales or purposes due to the occurred. service representative. system error. Restart the system again. ERROR...
Page 215 0x910A circuit may have stopped because the leak occurred. sensor cannot be Switch the instrument power recognized correctly. OFF and contact a Shimadzu FATAL sales or service representative. LEAK SENSOR A broken circuit Pump: The pumping was 0x910B may have occurred stopped because leak in the leak sensor.
Page 216 Restart the instrument OFF and contact a Shimadzu and start the pumping sales or service representative. again. ERROR LEAK DETECTED Solvent delivery was...
Page 217: Autosampler
If the above measures do not resolve the problem, switch the instrument power OFF and contact a Shimadzu sales or service representative. FATAL Switch the power OFF and SYSTEM ERROR A system error...
Page 218 0x5205 movement or there position of the needle X line of movement, contact a is a mechanical shaft (left-right direction) Shimadzu sales or service problem in the drive failed to be detected. representative. unit. Check for any obstacles. ERROR...
Page 219 0x520C disconnected or If the error is not resolved, information of the there is an electrical contact a Shimadzu sales or high-pressure valve were contact problem. service representative. not able to be read. Check if the high-pressure valve is disconnected.
Page 220 0x5210 disconnected or If the error is not resolved, information of the there is an electrical contact a Shimadzu sales or low-pressure valve were contact problem. service representative. not able to be read. Check if the low-pressure valve is disconnected.
Page 221 OFF and the temperature temperature. Pull out the contact a Shimadzu sales or inside the cabinet is rack and ventilate the service representative. outside the specified cabinet. range.
Page 222 OFF and contact a Shimadzu error for a power fan in sales or service representative. the back. Check the fan for obstacles.
Page 223 If the above measures do not resolve the problem, switch the instrument power OFF and contact a Shimadzu sales or service representative. FATAL The leak sensor LEAK SENSOR cannot be detected Switch the instrument power properly.
Page 224 Autosampler: The encoder is not counting again. If the error is not sensor of needle Y shaft 0x5222 normally in the resolved, contact a Shimadzu (front-rear direction) is needle Y-axis sales or service representative. not operating normally. (front-rear) direction. Press the Error Clear button to retry.
Page 225 [Power Install the front panel button] to switch OFF the properly. power supply and contact a Shimadzu sales or service representative. ERROR ADJUST PLATE The needle position Autosampler: Analysis was was not adjusted...
Page 226 Optimize the program so that exceeded the limit the number of lines of the 0x5233 value (max. 255 Autosampler: pretreatment program is bytes per file or Pretreatment program has reduced. 4,096 bytes for the an error. entire program). Nexera CL series...
Page 227 Change the setting to a value 0x5239 PURGE.ML command or PURGE.ML command in within the specified range. in the pretreatment the pretreatment program program exceeds is out of range. Change the setting range. the liquid volume to within the range. Nexera CL series...
Page 228 Change the setting to a value 0x523E pretreatment program for pretreatment within the specified range. the dispense is out of program exceeds range. Change the the setting range. dispense speed to within the range. Nexera CL series...
Page 229 MIX command Change the setting to a value 0x5243 command of the in the pretreatment within the specified range. pretreatment program is program exceeds out of range. Change the the setting range. setting to within the range. Nexera CL series...
Page 230 APPORT command Change the setting to a value 0x5248 command of the in the pretreatment within the specified range. pretreatment program is program exceeds out of range. Change the the setting range. setting to within the range. Nexera CL series...
Page 231 Autosampler: The wait Change the setting to a value 0x524D pretreatment time in the pretreatment within the specified range. program exceeds program is out of range. the setting range. Change the wait time to within the range. Nexera CL series...
Page 232 Revise the line number for the the FOR command and 0x5252 line number that is GOTO command. NEXT command is within a FOR-NEXT specified with the GOTO range. command. Correct the line number of the GOTO command. Nexera CL series...
Page 233 Autosampler: The wait Change the setting to a value 0x5257 pretreatment time in the pretreatment within the specified range. program exceeds program is out of range. the setting range. Change the wait time to within the range. Nexera CL series...
Page 234 0x525C than 19 IF Set it to 19 or fewer. more than 19 IF commands in its IF commands in its IF command block. command block. Make the number of IF commands 19 or less. Nexera CL series...
Page 235 FOR pretreatment command of the 0x5261 Enter the correct variable. program does not pretreatment program match the variable does not match the for the NEXT variable in the NEXT command. command. Input a correct variable. Nexera CL series...
Page 236 PFILE ASSIGN LVAR assigned by an Autosampler: The assignment variables assigned to the 0x5266 statement in the Enter the correct variable. assignment statement of pretreatment the pretreatment program program is not are not correct. Input correct. correct variables. Nexera CL series...
Page 237 0x526A movement or there position of arm rotation line of movement, contact a is mechanical direction failed to be Shimadzu sales or service problem in the drive detected. Check for any representative. unit. obstacles. ERROR...
Page 238 Plate changer: The arm 0x526D movement or there line of movement, contact a Z-axis (up-down direction) is mechanical Shimadzu sales or service motor slipped. Check for problem in the drive representative. any obstacles. unit. ERROR...
Page 239 OFF and temperature control was the temperature contact a Shimadzu sales or stopped. Open the door inside the cabinet is service representative. and ventilate the cabinet. outside the specified range.
Page 240 OFF and temperature control was the temperature contact a Shimadzu sales or stopped. Open the door inside the cabinet is service representative. and ventilate the cabinet. outside the specified range.
Page 241 Autosampler: The Rotary detected from the 0x5282 switch the power OFF and Valve 1 in the FCV1 unit connected contact a Shimadzu sales or autosampler has detected to the autosampler. service representative. an error. Check Rotary Valve 1. ERROR FCV2 ERROR Check the FCV2 unit.
Page 242 Autosampler: The Rotary detected from the 0x5285 switch the power OFF and Valve 4 in the FCV4 unit connected contact a Shimadzu sales or autosampler has detected to the autosampler. service representative. an error. Check Rotary Valve 4. ERROR FCV5 ERROR Check the FCV5 unit.
Page 243 0x5F02 operation panel that functionality for restoring settings were not set replacement. lost data cannot be used. correctly. The backup Contact your Shimadzu function is not working representative. properly. Nexera CL series...
Page 244 If it occurs frequently, switch The instrument was 0x5F05 reasons, because a the instrument power OFF and restarted for safety system error contact a Shimadzu sales or purposes due to the occurred. service representative. system error. Restart the system again. FATAL...
Page 245 Main Cause Countermeasure Code Lower: Workstation and Controller Screens FATAL The backup Contact our sales office / PCGx BACKUP parameter of PCGx 0x85E9 distributor or our designated could not be Rack changer system error service representative. acquired. Nexera CL series...
Page 246: Column Oven
1 "0x##" indicates a instrument power OFF and is restarted because an valve unit error. contact a Shimadzu sales or error is detected in the Refer to the valve service representative. valve. unit instruction manual.)
Page 247 The leak sensor cannot be detected Switch the instrument power Column oven: The 0x5405 properly. A broken OFF and contact a Shimadzu temperature control was or short circuit may sales or service representative. stopped because the leak have occurred. sensor cannot be correctly recognized.
Page 248 ROOM TEMP SENSOR A broken or short circuit may have Switch the instrument power Column oven: The 0x5409 occurred in the OFF and contact a Shimadzu temperature control was room temperature sales or service representative. stopped because the sensor. room temperature sensor cannot be read.
Page 249 If it occurs frequently, switch The instrument was 0x5F05 reasons, because a the instrument power OFF and restarted for safety system error contact a Shimadzu sales or purposes due to the occurred. service representative. system error. Restart the system again. FATAL...
Page 250 FATAL THERMAL FUSE A broken circuit Switch the instrument power Column oven: 0x9404 may have occurred OFF and contact a Shimadzu Temperature fuse was in the thermal fuse. sales or service representative. disconnected, and the cell temperature control was stopped.
Page 251 Remove anything that contact a Shimadzu sales or interferes with the service representative. rotation of the fans and restart the instrument. FATAL...
Page 252 FATAL AUX HEATER The auxiliary heater Switch the instrument power Column oven: The 0x9410 is not functioning OFF and contact a Shimadzu temperature control was properly. sales or service representative. stopped because the auxiliary heater did not heat up. ERROR...
Page 253: Uv Detector
To replace the current DET UV detector: Detector 0x5803 board is not board with one for a micro board for micro cell properly recognized. cell, contact a Shimadzu sales cannot be recognized or service representative. correctly. ERROR LEAK DETECTED Inspect the detector leak and...
Page 254 CELL TEMP SENSOR A broken circuit Switch the instrument power UV detector: Temperature may have occurred 0x5870 OFF and contact a Shimadzu of the flow sensor did not in the cell sales or service representative. operate normally, so cell temperature sensor.
Page 255 0x58C4 UV detector: W Lamp or the illumination the instrument power OFF and does not light. Replace circuit is not contact a Shimadzu sales or the W Lamp. functioning properly. service representative. WARNING The lamp housing LAMP COVER OPEN Close the lamp housing cover.
Page 256 Shimadzu sales or service representative. FATAL FILTER POSITION Detection of the Switch the instrument power 0x58D1 filter motor home OFF and contact a Shimadzu UV detector: Initialization position failed. sales or service representative. was not able to finish normally. ERROR The illumination...
Page 257 Code Lower: Workstation and Controller Screens FATAL DETECTOR Initialization of Switch the instrument power 0x58DA internal actions OFF and contact a Shimadzu UV detector: Initialization failed. sales or service representative. was not able to finish normally. FATAL DETECTOR Initialization of...
Page 258 MIRROR POSITION motor home Switch the instrument power UV detector: Initialization position failed for 0x58F7 OFF and contact a Shimadzu was not able to finish no the lamp switching sales or service representative. UV detector: Initialization mirror control was not able to finish motor.
Page 259: Selecting Components
UHPLC system with lower capacity, but capable of ultra-fast High-pressure gradient multianalyte analysis or applications Ultra High-Speed Analysis system with an MS front end, using a column with 2 µm or smaller particles. Nexera CL series...
Page 260: System Installation Arrangement
High-Pressure Gradient Systems (High-Speed Analysis)* Name Model Qty. System Controller CBM-40 CL Solvent Delivery Unit LC-40B X3 CL Degassing Unit Autosampler SIL-40C X3 CL Column oven CTO-40C CL Detector SPD-40 CL Reservoir Tray Mixer MR 40 µL mixer One possible example Nexera CL series...
Page 261 CBM-40 CL Solvent Delivery Unit LC-40D XR CL Low-Pressure Gradient Kit Degassing Unit DGU-405 CL Autosampler SIL-40C XR CL Column oven CTO-40C CL Detector SPD-40 CL Reservoir Tray Mixer MR 300 µL LPGE mixer One possible example Nexera CL series...
Page 262: System Tubing
Male Nut, 1.6 MN Male Nut, 1.6 MN W6 UHPLC Fitting 2, S Ferrule, 1.6F 316L Coupling, 1.6X-0.4 Instruction Manual Connected in place of a column when a column is not used, such as while rinsing flow channels. Nexera CL series...
Page 263 6.1 Selecting a System An example of the analysis flow channel tubing connections for a typical configuration is shown below. (The indicated numbers correspond to numbers in the Part Details table on the previous page.) Nexera CL series...
Page 264: Ultra High-Speed Analysis
Overlapping Injection However, if it affects the baseline, clear the checkbox. The overlapping injection function is only enabled for continuous analysis using the same method. If adequate reproducibility is achieved, then shorten the Time Program equilibration time. Nexera CL series...
Page 265: Selecting A Mixer
If multiple detectors are connected, use the mixer with the largest capacity. For example, if using LC/MS and Absorbance Detector with Containing Trifluoroacetic Acid as mobile phase, use the MR180 µLII. If the type of mobile phase to be used is unknown, use the largest capacity mixer, which is the MR180 µLII. Nexera CL series...
Page 266: Selecting The Cell
Semi-micro cells have a small 2.5 µL volume, but the optical path length is 5 mm, which results in about half the peak intensity of a UHPLC cell. To reduce noise for analytical conditions with flowrates of 1.5 mL/min or higher, use a conventional cell. Nexera CL series...
Page 267: Installation
Does not include materials/devices for Column Oven CTO-40C CL securing the unit during shipping. Remove protective tape, etc. CBM-40 CL or Reservoir Tray Does not include materials/devices for securing the unit during shipping. Nexera CL series...
Page 268: Installing The Autosampler
• Install the autosampler at the bottom level of the system. • Horizon length of the front of autosampler from the end of the lab table should be limited within 100mm. Long horizontal length may prevent waste liquid from being smoothly discharged. Nexera CL series...
Page 269 Install caps firmly all the way in. They could contact the Z-mount if they protrude. Attach rinse port cover (A) to rinse port. Attach the cover securely all the way in. If lifted, it may come in contact with the needle mount. Nexera CL series...
Page 270 For more detailed front panel installation/removal instructions, refer to the SIL-40 CL Series Instruction Manual. Refer to "8.2 Installing/Removing Front Panels" in the SIL-40 CL Series Instruction Manual. Install the upper front panel (C) included in accessories. Install the lower front panel (D) included in accessories. Nexera CL series...
Page 271: Installing The Solvent Delivery Pump
7.1 Installing the System Insert the sample rack (E) included in accessories. Contact a Shimadzu sales or service representative to have the plate changer installed. Plate changer installation involves operations with the main unit cover removed, which is dangerous. Plate changers can only be installed on the left side of a SIL-40 CL series autosampler.
Page 272: Installing The Detectors (Spd-40 Cl)
This hole is used to insert detector inlet tubing into a column oven or other unit installed on the right side. If the cap is difficult to remove, then use a tool or other hard object to push on the cap. Nexera CL series...
Page 273: Installing The Degassing Unit (Dgu-405 Cl)
Installing the Right Panel Insert the two tabs on the right side of the right panel into the holes in the main frame. Fasten the right panel by engaging the latch (1). Nexera CL series...
Page 274: Installing The Reservoir Tray
Insert the tray back into the main unit. Insert the tabs protruding from the front of the tray into the holes in the back side of the front panel. Refasten and hand tighten the knurled screw (A) that was removed in step 1. Nexera CL series...
Page 275: Installing The Column Oven (Cto-40C Cl)
CTO-40C CL cover. It cannot measure room temperature accurately if it is touching the cover. n Installing the Dust Filter (Included) Insert the dust filter (B) into the included filter holder (A). Nexera CL series...
Page 276: Installing Other Instruments
Before installing the column oven, connect a drain tube for draining liquid waste. Connecting the Drain Tube to the Column Oven (CTO-40C CL) 7.1.8 Installing Other Instruments For information about installing other instruments, refer to the instruction manual included with respective instruments. Nexera CL series...
Page 277: Cable Connections
7.2.2 Connecting Optical Cables n How to Connect Optical Cables Connect the optical cable, which is a two-conductor cable that sends and receives signals via the same cable, to the [REMOTE] connector. Nexera CL series...
Page 278 Do NOT insert the optical cable plug with the plug overlapping both channels. Doing so could cause an equipment failure or communication problem. Grip the plug to connect or disconnect the cable. Gripping the cable could damage the plug or cause a broken circuit in the cable. Nexera CL series...
Page 279 [0] (default value) in the system setting group. Confirm that the [CONTROLLER LINK] setting for the solvent delivery pump is [1] (the default value is the external connection mode) in the system setting group. Nexera CL series...
Page 280 A. For example, if [LINK ADDRESS] is set to 4, then the address for pump A is set to 4 and the address for pump B is set to 5. Nexera CL series...
Page 281: Connecting A Degassing Unit (Dgu-405 Cl) To The Solvent Delivery Pump
(vacuum level output) is connected. If so, connect a ground cable to the pump or detector as shown. Only loosen the screw enough to insert the crimped terminal. The screw can be difficult to screw back in if completely removed. Nexera CL series...
Page 282 7 Installation Label or Name GND Cable Crimped Terminal (on DGU unit) Main Unit Ground Terminal Screw Crimped Terminal (on pump/detector unit) Nexera CL series...
Page 283 Secure the cables by installing two covers (C). Install the covers by sliding them up or down. A clamp (072-60658-01) is packed in the cable kit for a spare part. Use it for arranging the cables as necessary. Nexera CL series...
Page 284: Tubing Connections
The reservoir bottle described under optional parts is recommended for the reservoir bottle. To use a different reservoir bottle, prepare a glass bottle with at least a 500 mL volume. Do NOT use a cracked or scratched bottle. Doing so could cause bottle damage. Nexera CL series...
Page 285 Prepare tanks for liquid waste and for liquid waste at risk of infection. Tubing with no risk of infection can become contaminated. Put the label on liquid waste tanks that have a risk of infection to distinguish them. Nexera CL series...
Page 286: Installing Tubes And Joints For Draining Leakage
• To improve liquid waste flow, orient the end of the silicone tubing downward. • Cut the silicone tubing to a length that prevents sagging. Nexera CL series...
Page 287 Silicone rubber tubing, O.D 8.0 ´ I.D. 6.0 016-31350-02 mm, natural color 228-66695 Drain tube Insert the drain tubes firmly all the way. If it is not inserted all the way, fluid may leak from the connection. Nexera CL series...
Page 288 • Cut the drain tube to an appropriate length that prevents sagging. • The connection method varies depending on the type of liquid waste bottle. Read the precautions in the instruction manual included with the drain tube and select the appropriate connection method. Nexera CL series...
Page 289 (A). • When the drain tube is secured, make sure the internal diameter is not flattened. • Position the drain tube in a straight line so that it does not bend. Nexera CL series...
Page 290 Cut the silicone tube (1) at the edge of the table and connect the included elbow joint. Orient the elbow joint downward and connect the silicone tube. Insert the opposite end of the silicone tube into the liquid waste bottle. Nexera CL series...
Page 291 Column ovens are designed so that leakage from instruments above does not flow down. From the front side, insert the included CTO drain outlet tube (1) into the leakage drain port. Rotate the CTO drain outlet tube counterclockwise 45 degrees. That secures the CTO drain outlet tube. Nexera CL series...
Page 292 Connect the included elbow joint (A) and silicone tube (C) to the drain port. To connect multiple plate changers, use the included Y-joint (B) to merge flow channels. Routing the Tube Behind the Installation Routing the Tube in Front of the Installation Base Base Connecting the Plate Changer to Multiple Units Nexera CL series...
Page 293 Use a tube clamp to fasten the condensation drain tube to the liquid waste bottle. n Connecting the Solvent Delivery Pump Drain tube Cut the drain tube to an appropriate length and fix the end to the tank for liquid waste having no risk of infection. "Fastening Tubing" P.292 Nexera CL series...
Page 294 Insert the end of the drain tube into the autosampler drain port and close the clamps on the right panel to secure the tube. Adjust the tube length so that it can be inserted into the autosampler drain block. "Fastening Tubing" P.292 Nexera CL series...
Page 295: Connecting Low-Pressure Flow Channel Tubing
Use suction filter and tubing included with autosamplers. Use tubing included with autosamplers, cut to an appropriate length (1.6 mm O.D. ´ 0.5 mm I.D.). Use flangeless fittings included with autosampler units. Connected to autosampler low-pressure valve (port 3) or multi-rinsing inlet. Nexera CL series...
Page 296 Plumbing Procedure for the degassing unit liquid inlet/outlet ports Slide the nut(A), lock ring(B) and ferrule(C), in that order, onto the tubing. The flattened end(a) of the ring should face towards the nut(A) with the narrow end(b) of the ring towards the nut. Nexera CL series...
Page 297 Before connecting the tubing, remove the stop joints that are installed in the degassing unit liquid inlet/outlet ports. (Leave the stop joint installed for unused flow channels.) Nexera CL series...
Page 298 Use suction filter and tubing included with autosamplers. Use flangeless fittings included with autosampler units. Use tubing included with autosamplers, cut to an appropriate length (1.6 mm O.D. ´ 0.5 mm I.D.). Connected to autosampler low-pressure valve (port 3) or multi-rinsing inlet. Nexera CL series...
Page 299 Use joints included with the autosampler to connect to the autosampler low-pressure valve inlet (port 3) or the multi-rinsing inlet. Rinse solution purging time settings can be shortened by using a rinse solution suction tube that is as short as possible. Nexera CL series...
Page 300 • Do not make the FEP tubing from the degassing unit outlet to the low-pressure gradient unit inlet too long. Unnecessarily long FEP tubing can cause air to be reabsorbed in the tubing. Nexera CL series...
Page 301 Cut the suction tube to the necessary length. Place the mobile phase and rinse solution solvent in reservoir bottles. Insert the filter portion into the reservoir bottle. Insert the filter portion all the way to the bottom of the reservoir bottle. Nexera CL series...
Page 302 Use bushings (B) and ferrules (C) to connect the tubing from the degassing unit to the pump inlet. To orient the tubing exiting the suction filter upward, secure it with the tubing holder. Nexera CL series...
Page 303 Connect the rinse solvent tube to LPV port #3 (A) with a mail nut. In case of SIL-40C X3 CL Connect 3 tubes for rinse solvent to R0-R2 (A) of the multi-rinse block using mail nuts and PTFE ferrules. Nexera CL series...
Page 304 DGU-405 CL port farthest to the right. Use the flangeless fittings and ferrules included with the DGU-405 CL or LC-40B X3 CL unit to connect the suction filter assembly included with the autosampler to the degassing unit inlet port. Nexera CL series...
Page 305 Remove the front cover (A) from the reservoir tray. Affix the label (b) to the tube clamp (a). Fasten suction tube (D) to the guide bar (C) using the tube clamp (a). Return the front cover (A) to its original position. Nexera CL series...
Page 306 Then insert the front cover latch into the instrument (B). n Fastening Tubing Use the tubing holder to clamp the tubing that passes to the right of the solvent delivery pump and detector. Nexera CL series...
Page 307 Do not insert the drain tube of detector to the inside of the drain port (a). There is a risk of deterioration of drainage. Fix the solvent delivery pump drain tube to the tank for liquid waste having no risk of infection. Nexera CL series...
Page 308: Connecting Tubing For High-Pressure Flow Channels (Connection Methods)
Tubing Kits Four types of tubing kits, which contain a set of tubing and fittings optimized for typical system configurations, are available for Nexera CL series systems. For more details about tubing kits, refer to the instruction manual for tubing kits.
Page 309 Insert tubing in until it hits the back of the connection hole. A tube that does not reach the back of the connection hole creates dead volume that could cause peak broadening. Tighten the male nut hand-tight. Nexera CL series...
Page 310 Connecting UHPLC Fittings UHPLC fittings have a pressure capacity of 130 MPa. They also can be removed from tubing and reused even if they were previously tightened. UHPLC fittings are used for connecting column inlet tubing. Nexera CL series...
Page 311 Insert the tubing all the way in, until it hits the back of the connection hole. If the tubing does not reach the back of the connection hole, it creates dead volume that could cause peak broadening. Nexera CL series...
Page 312 228-56866 specialized tool for UHPLC fittings). a) Using UHPLC Fittings for the First Time Tighten the nut an additional 180 degrees (half turn) from the hand-tightened position. Use the slots in the nut as a reference point. Nexera CL series...
Page 313 Insufficient tightening might cause leakage when solvent is delivered at 130 MPa. Do NOT tighten more than 180 degrees (half turn) from the hand-tightened position. Excessive tightening could deform the fitting or scratch/gouge the tubing, which could decrease the number of available reuses. Nexera CL series...
Page 314: Connecting Tubing For High-Pressure Flow Channels (Gradient Systems)
If the column oven is operated at temperature settings above 85 °C, either use the mixer which is disconnected from the recognition device (CMD) or use the mixer without the mixer recognition device. For information about installing other mixers, contact a Shimadzu representative to confirm the installation procedure. Nexera CL series...
Page 315 Remove the cap (A) from the CMD connector on the main column oven unit. The step is not necessary if the mixer recognition function is not used. Install the mixer (A) inside the column oven using the screws (B) included with the mixer. Nexera CL series...
Page 316 Attach a 1.6 MN male nut and ferrule included with the tubing kit to the opposite end of the stainless steel tubing (0.3 ´ 600 mm), which was connected to the pump, and connect that opposite end to the mixer inlet. Nexera CL series...
Page 317 Connecting Tubing from the Detector Outlet to the Drain "Connecting the Detector Drain tube" P.280 n Other Tubing Connections For information about connecting tubing for other instruments, refer to the instruction manual provided with respective instruments. Nexera CL series...
Page 318: Connecting Tubing For High-Pressure Flow Channels (Isocratic Systems)
Use a W6 male nut and ferrule to connect the opposite end to port 4 on the autosampler high-pressure valve. Use the same procedure as described in "7.3.4 Connecting Tubing for High-Pressure Flow Channels (Gradient Systems)" P.300 to connect tubing from the autosampler to the column and from the column to the detector. Nexera CL series...
Page 319: Checking Computer And System Controller Connections
Do NOT connect two or more system controllers with factory settings to the LAN at the same time. Doing so will cause an IP address conflict. Connect one device to the LAN at a time and change the IP address. Nexera CL series...
Page 320: Setting The Computer Ip Address
On the Start menu, click [Settings]-[Network & Internet]. Click [Change your network settings]-[Change adapter options]. Right-click [Ethernet] and then click [Properties] on the right-click menu. In the [Ethernet Properties] window, select [Internet Protocol Version 4 (TCP/IP)] from the list and click the [Properties] button. Nexera CL series...
Page 321 7.4 Checking Computer and System Controller Connections Specify the [IP address] and [Subnet mask] settings. When finished, click [OK]. If prompted to do so, restart the computer. Nexera CL series...
Page 322: Specifying Computer Browser Settings
On the [Security] tab page, click [Custom level]. Configure settings as indicated in the table below. Item Setting [Run ActiveX controls and plug-ins] [Enable] [Script ActiveX controls marked safe for scripting] [Enable] [Active Scripting] [Enable] [Enable] or [Access data sources] [Prompt] Nexera CL series...
Page 323 Click [Advanced]. Then add the URL address for the system controller (“192.168.200.*”) in the [Exceptions] field. Click [OK]. Specify [Advanced] tab page settings. Click the [Advanced] tab. In the [Settings] field, select the [Browsing]-[Display a notification about every script error] checkbox. Nexera CL series...
Page 324: Specifying System Controller Settings
Click the (Administrator) button in the upper right area of the [Device Monitor] window and log in to the [Group Management] window. When shipped with factory settings, log in by entering “Admin” as the [User Name] and [Password]. Nexera CL series...
Page 325 Click [Use the following IP Adress] and then specify the [IP Adress] and [Subnet Mask] settings. Clicking the [Advanced Settings] button displays the [Advanced Settings] window, so that the [Default Gateway] setting can be specified. Nexera CL series...
Page 326: Other Installation Processes
Connect the straight LAN cable to the Ethernet connector on the back of the CBM-40 CL unit. Connect the other end of the straight LAN cable, which was connected to the system, to the switching hub that is connected to the computer. Nexera CL series...
Page 327: Installing Optional Column Management Functionality
For more details, refer to the instruction manual for the LabSolutions chromatography data system. Part Name CMD (Column Management Device) 228-37281-41 CMD cable (150 mm) 228-39991 CMD cable (400 mm) 228-39991-01 CMD cannot be used at temperature settings above 85 ℃. Nexera CL series...
Page 328 To connect a column with a length of 100 mm or less, use a 400 mm long CMD cable (228-39991-01). Remove the cap (A) from the CMD connector on the main column oven unit. Connect the end of the CMD cable (B) to the CMD connector (C) on the main column oven unit. Nexera CL series...
Page 329 CMD cable during analysis. Doing so could corrupt the data. Install the column with the CMD attached into the column oven, as described in "2.4.1 Attaching Columns in the Column Oven" P.29. "2.4.1 Attaching Columns in the Column Oven" P.29 Nexera CL series...
Page 330: Addendum
In ion pair chromatography, “p'+0.25e” indicates the dielectric constant as a (11) function of p', which is proportional to mobile phase strength. A.M. Krstulovic, P.R. Brown: Reversed-Phase High-Performance Liquid Chromatography (1982) Sources Wiley Interscience: Translated by Makino et al, Reversed-Phase High-Performance Liquid Chromatography (1985), Tokyo Kagaku-Dojin Publishing Nexera CL series...
Page 331 1.521 0.75 Bromobenzene 1.557 1.04 Iodobenzene Phenyl ether 1.58 Phenetole 1.505 1.14 Ethyl ether(**) 1.35 0.24 Benzene 1.498 Tricresyl phosphate Ethyl iodide 1.51 0.57 n-Octanol 1.427 Fluorobenzene 1.46 0.55 Benzylether 1.538 Methylene chloride(**) 1.421 0.41 Anisole 1.514 Nexera CL series...
Page 332 0.35 i-Propyl ether(*) 0.28 0.62 Toluene 0.29 0.046 p-Xylene 0.26 Chlorobenzene Bromobenzene 0.32 Iodobenzene 0.35 Phenyl ether Phenetole Ethyl ether(**) 0.38 Benzene 0.32 0.058 Tricresyl phosphate Ethyl iodide n-Octanol 10.3 Fluorobenzene Benzylether Methylene chloride(**) 0.42 0.17 Anisole Nexera CL series...
Page 333 1.507 0.88 Nitroethane 1.39 0.64 Acetone(*) 1.356 Benzyl alcohol 1.538 Tetramethyl guanidine Methoxyethanol Tris(cyanoethoxy) propane Propylene carbonate Ethanol 1.359 10.8 Oxydipropionitrile Aniline 1.584 3.77 Acetic acid 1.37 Acetonitrile(*) 1.341 0.34 N,N-dimethylaceta-mide 1.436 0.78 Dimethylformamide 1.428 Dimethylsulfoxide 1.477 Nexera CL series...
Page 334 12.4 Nitroethane Acetone(*) 0.71 miscible 13.1 Benzyl alcohol Tetramethyl guanidine Methoxyethanol miscible 19.9 Tris(cyanoethoxy) propane 0.56 Propylene carbonate Ethanol miscible 24.6 Oxydipropionitrile Aniline Acetic acid miscible Acetonitrile(*) miscible 37.5 N,N-dimethylaceta-mide 0.88 37.8 Dimethylformamide 36.7 Dimethylsulfoxide 0.62 miscible Nexera CL series...
Page 335 (℃) (**)η ≦ 0.5 cp, B.P. < 45 ℃ N-methyl-2-pyrolidone 1.468 1.67 Hexamethyl phosphoric 1.457 acid triamide Methanol(*) 1.326 0.54 Nitromethane 1.38 0.61 m-Cresol 1.54 N-methylformamide 1.447 1.65 Ethylene glycol 1.431 16.5 Formamide 1.447 Water 1.333 0.89 Nexera CL series...
Page 336 (*)η ≦ 0.5 cp, B.P. > 45 ℃ Constant e°a 20 ℃ 0.25e (**)η ≦ 0.5 cp, B.P. < 45 ℃ Solvent N-methyl-2-pyrolidone Hexamethyl phosphoric acid 0.65 triamide Methanol(*) miscible 32.7 Nitromethane m-Cresol 11.8 N-methylformamide miscible Ethylene glycol miscible 37.7 Formamide miscible Water 10.2 Nexera CL series...
Page 337: Index
Testing the Temperature Adjustment Function 137 Inspection Environment Timing for Hardware Validation Inspection Equipment Tubes and Joints for Draining Leakage Instruction Manual Components Tubing Connections Isocratic Systems UHPLC Fittings LabSolutions 12, 15, 312 List of Parts Required for Inspection Nexera CL series...
Page 338 When Replacing the Mobile Phase Nexera CL series...

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