Nexera HPLC Troubleshooting the Autosampler

Browser lacks capabilities required to play.

Upgrade or switch to another browser.

Loading…

<div class="container"><h1 class="title">Troubleshooting the Autosampler</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/power-indicator.jpg" alt="Troubleshooting the Autosampler"></div> <h3>Examine the power indicator on the front of the autosampler.</h3> The power indicator bar is located on the right side of the display panel. <ul> <li>Green indicates that the power is ON.</li> <li>Red indicates that an error has been generated.</li> <li>Orange indicates that the unit is in sleep mode.</li> </ul> <div class="ts-question">Is the indicator illuminated?</div><div class="buttons"> <span class="btn-yes">[[Yes->Was an Injection Made?]]</span> <span class="btn-no">[[No->Power Connections]]</span> </div> </div>

Config.debug = false;

<div class="container"><h1 class="title">Was an Injection Made?</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/solvent-peak.jpg" alt="Was an Injection Made?"></div> By closely examining the first few minutes of the chromatogram, it can be determined whether an injection is being made. The injection process typically produces a baseline disturbance caused by the detectors response to: <ul> <li>The sample diluent.</li> <li>A pressure fluctuation from the rotation of the injection valve.</li> <li>A change in the refractive index.</li> </ul> The retention time of this disturbance is indicative of the column volume and is referred to as the void time (time=0). If there is no indication of a successful injection, the problem is most likely with the signal cable, the injection process or the detector. If the solvent peaks are present, it is an indication that something was injected and the detector is responding to it. If there is a chromatography problem, it may be related to the injector, detector, or sample solution. <div class="ts-question">Are there any peaks in the chromatogram?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Peak Shape]]</span> <span class="btn-no">[[No->Sample Vial Position]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Power Connections</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/power.jpg" alt="Power Connections"></div> <h3>Examine the connections of the power supply cord.</h3> One end of the power cable is inserted into the power connector on the back of the autosampler. The other end of the power cable is inserted into the power supply outlet. <img class="warning" style="float: left; margin-bottom:25px;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: Ensure that the power switch is in the OFF position before connecting the power cable. WARNING: The three-line type power cable supplied as an accessory includes the grounding wire. Connect this wire to ground to prevent electrical shock and to secure stable operation of the instrument.</strong> <div class="ts-question">Is the power cord properly connected to the autosampler and power supply outlet?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Power Cord Damage]]</span> <span class="btn-no">[[No->Making the Power Connection]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Peak Shape</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/peak-shape.jpg" alt="Peak Shape"></div> The most optimal peak shape is a Gaussian curve. A Gaussian curve is a perfectly symmetrical curve that represents a normal distribution pattern. Compare the current analyte peak shape to the established historical peak shape. Determining whether the current chromatogram is equivalent to the historical peak shape allows us to further isolate the problem and find an effective corrective action. The tailing factor is the most common way to determine whether the peak shape is optimal. Most computer-based chromatography software applications can automatically calculate the tailing factor. Consult the software documentation or use the formula above to calculate the tailing factor. Tailing Factor=1.0 is perfect. <div class="ts-question">Is the shape of the sample peaks normal?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Fluctuating Retention Time]]</span> <span class="btn-no">[[No->Autosampler Tubing]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Sample Vial Position</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/vial-position-sil.jpg" alt="Sample Vial Position"></div> The upper surface of the sample vial rack is marked with numbers at the vial positions. These numbers correspond to the sample vial positions in the chromatography software. Ensure that the sample vial is actually in the position designated in the injection sequence. <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>NOTE: The vial position numbers start with 1. Ensure that the sample vial is actually in the position designated in the injection sequence and that the proper rack type is designated.</strong> <div class="ts-question">Has the sample vial been correctly placed into the sample vial rack?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Sample Vial Volume]]</span> <span class="btn-no">[[No->Vial Position in the Autosampler]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Power Cord Damage</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/power-cord-damage.jpg" alt="Power Cord Damage"></div> Handle the power cord with care. Observe the following precautions to avoid the possibility of cord damage that may result in fire, electric shock or instrument malfunction. <ul> <li>Do not place heavy objects on the cord.</li> <li>Keep hot items away from the power cord.</li> <li>Do not modify the power cord in any way.</li> <li>Do not bend the cord excessively.</li> <li>To unplug the cord, pull on the plug itself, NOT on the cord.</li> <li>If the cord becomes damaged, replace it with a new one immediately.</li> </ul> <div class="ts-question">Is the power cord damaged?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Power Cord Replacement]]</span> <span class="btn-no">[[No->Power Requirements]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Making the Power Connection</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/power.jpg" alt="Making the Power Connection"></div> <ol> <li>Turn the power switch on the detector OFF before plugging the instrument into the power supply.</li> <li>Insert the plug end of the power cable directly into the power supply outlet. </li> <li>Insert the connector end of the power cable into the power connector on the back of the detector.</li> </ol> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>WARNING: The three-line type power cable supplied as an accessory includes the grounding wire. Connect this wire to ground to prevent electrical shock and to secure stable operation of the instrument.</strong> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Fluctuating Retention Time</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/retention-time-change.jpg" alt="Fluctuating Retention Time"></div> <h3>Compare current chromatography with historical results.</h3> Several injections must be observed to diagnose a retention time problem. Different patterns in the retention times indicate different diagnoses. <ul> <li>Drifting (shorter retention times) - indicates column degradation.</li> <li>Drifting (longer retention times) - may indicate a steadily decreasing column temperature.</li> <li>Irregular - indicates a random shifting of the flow rate of the analytes through the column matrix.</li> <li>Steady but Different - indicates a change in the volume of the system flow line.</li> </ul> <div class="ts-question">Are the retention times of known peaks fluctuating?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Waste Discharge (1595)]]</span> <span class="btn-no">[[No->Ghost Peaks (1610)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Autosampler Tubing</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/flow-sil.jpg" alt="Autosampler Tubing"></div> The tubing into and out of the autosampler may cause some confusion. The inlet tubing brings mobile phase from the pump or mixer into port 6 of the high-pressure valve.This inlet tubing IS marked with a blue band. The outlet tubing carries the mobile phase and sample from port 5 of the high-pressure valve to the column inlet. This outlet tubing IS NOT marked with a blue band. <div class="ts-question">Has any of the plumbing between the pump and column been reversed?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Autosampler (SIL) Plumbing]]</span> <span class="btn-no">[[No->Column Performance (1582)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Sample Vial Volume</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/vial-choice.jpg" alt="Sample Vial Volume"></div> Although the autosampler uses a direct volume injection with no sample loss, the minimum required sample volume is dependent on a number of factors. Consider all of these parameters when selecting the type of sample vial to be used and the final volume of sample in each vial. <ul> <li>Sample vial type</li> <li>Injection volume</li> <li>Needle stroke</li> </ul> It is also important to consider the maximum volume of sample in the sample vial. If the sample volume is too large, the sample may not cool (SIL-20AC only) properly. It is also possible that if the sample volume is too large, it could impede the sample aspiration process. <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>WARNING: The amount of sample introduced into the vial should not exceed the corresponding heights displayed in the diagram.</strong> <div class="ts-question">Is there a sufficient supply of sample in the sample vial?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Needle Stroke Depth]]</span> <span class="btn-no">[[No->Filling the Sample Vial]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Vial Position in the Autosampler</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/vial-position-sil.jpg" alt="Vial Position in the Autosampler"></div> <ol> <li>Open the door of the autosampler.</li> <li>Place the vial in the rack with the cap facing up.</li> <li>Slide the sample vial rack along the guides all the way to the back of the instrument. The sample vial rack clicks into place when it is inserted correctly.</li> <li>Close the door.</li> <li>Enter the proper sample vial positions into the injection sequence.</li> </ol> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>NOTE: The vial position numbers start with 1. Ensure that the sample vial is actually in the position designated in the injection sequence and that the proper rack type is designated.</strong> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Needle Stroke Depth</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/needle-stroke-depth.jpg" alt="Needle Stroke Depth"></div> The distance that the needle descends into the sample vial is determined by the NEEDLE STROKE parameter. The parameter resets to the default position (2 mm from the bottom of the vial) when the sample vial rack is changed. This parameter must be adjusted for each of the different rack types. <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>NOTE: The table above displays the default values for needle stroke.</strong> <div class="ts-question">Is the sample needle depth adequate for the vial type?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Autosampler Flow Line]]</span> <span class="btn-no">[[No->NEEDLE STROKE Parameter]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Filling the Sample Vial</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/filling-sample-vial.jpg" alt="Filling the Sample Vial"></div> Ensure that the final volume of sample in the vial is sufficient for the injection volume and the number of repeat injections. Verify that the sample solution does not contain particulate matter. Filter the sample or adjust the sample preparation procedure to ensure adequate sample quality and volume. Ensure that the sample vial is properly covered and sealed to avoid sample evaporation. The table above displays the residual volume left in the bottom 2 mm of the vial <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Autosampler Flow Line</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/flow-sil.jpg" alt="Autosampler Flow Line"></div> The autosampler flow line is detailed above. Ensure that the flow lines are plumbed correctly. Please examine these flow lines and connections for blockage or leaks. <div class="ts-question">Are the flow lines in the injection flow path clogged?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Tubing Replacement (1545 / 1591)]]</span> <span class="btn-no">[[No->Check/Recheck Method]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">NEEDLE STROKE Parameter</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/IapCWjZ0UFk?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>NOTE: When the rack is changed, this parameter is automatically set to the default value. The default setting causes the needle to descend to 2 mm from the bottom of the sample vial.</strong> The NEEDLE STROKE parameter sets the distance that the needle descends into the sample vial. <ol> <li>From the Initial screen, press the FUNC key.</li> <li>The Parameter setting screen is displayed. Press the ENTER key.</li> <li>Then press the FUNC key seven times. The NEEDLE STROKE parameter screen is displayed.</li> <li>Enter the numeric value for the parameter and press the ENTER key.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Tubing Replacement</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/tubing.jpg" alt="Tubing Replacement"></div> <ul> <li>Use caution when replacing any tubing on the autosampler. </li> <li>Ensure that all tubing is replaced with tubing of the same internal diameter.</li> <li>Increasing the inner diameter of the tubing increases the volume of the flow path.</li> </ul> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Check/Recheck Method</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/sample-table.jpg" alt="Check/Recheck Method"></div> It is a very good practice to check and recheck the method even if this is a method that is run often. Things to recheck are listed below: <ul> <li>Column type and size</li> <li>Mobile phase composition and gradient conditions</li> <li>Flow rate</li> <li>Wavelength</li> <li>Sample preparation</li> </ul> Re-examine these items and verify that they are appropriate. <div class="ts-question">Are the injection parameters properly set?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Flow]]</span> <span class="btn-no">[[No->Changing the Injection Parameters]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Flow</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/waste.jpg" alt="Flow"></div> Examine the waste tubing as it enters the waste container. Is the flow of the discharge appropriate for the flow rate? <div class="ts-question">Is the mobile phase flowing?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Detector Function 1 (1558)]]</span> <span class="btn-no"><a href="https://www.ssi.shimadzu.com/service-support/virtual-advisor/nexera/advanced-troubleshooting/troubleshooting-the-pump/index.html">Troubleshoot the Pump</a></span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Changing the Injection Parameters</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/XX7kxC-RWV0?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>NOTE: If only one vial is to be injected, use the same number entered in the 'From' field.</strong> <ol> <li>From the Initial screen, press EDIT. The Editing Sample Table screen is displayed briefly.</li> <li>The cursor is active in the 'Rack' field.</li> <li>Using the numeric keypad, enter the rack number and press ENTER.</li> <li>The cursor moves to the 'From' field.</li> <li>Using the numeric keypad, enter the first vial number to inject, and press ENTER.</li> <li>The cursor moves to the 'To' field.</li> <li>Using the numeric keypad, enter the last vial to inject, and press ENTER.</li> <li>The cursor moves to the 'Rep' (repetitions) field.</li> <li>Using the numeric keypad, enter the number of times the autosampler will inject from the specified vial, and press ENTER.</li> <li>The cursor moves to the 'Inj Volume' field.</li> <li>Using the numeric keypad, enter the desired injection volume (µL) and press ENTER.</li> <li>The cursor moves to the 'Runtime' field.</li> <li>Enter the analysis time (min) for each injection, and press ENTER.</li> <li>The next line of the sample table is displayed.</li> <li>Press CE to return to the Initial screen.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Detector Function</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/detector-saturation.jpg" alt="Detector Function"></div> <h3>Press the appropriate button below to connect to troubleshoot your detector.</h3> It is possible that a problem with the detector may manifest itself as a problem with the autosampler. Press the appropriate button below to connect to troubleshoot your detector. <div class="ts-question">Is the detector functioning properly?</div> <span class="btn-link"><a href="https://www.ssi.shimadzu.com/service-support/virtual-advisor/nexera/advanced-troubleshooting/troubleshooting-pda-detector/index.html">Troubleshoot the PDA Detector</a></span> <div class="buttons"> <span class="btn-yes">[[Yes->Needle Position in Sample Vial]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Needle Position in Sample Vial</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/needle-position.jpg" alt="Needle Position in Sample Vial"></div> <h3>Examine the needle as it enters the sample vial.</h3> The ADJUST RACK parameter is used to adjust the needle position as it enters the vial opening. If the needle is not centered over the center of the vial opening, the needle may be damaged during an extended sample sequence. Examine the needle as it enters the sample vial. <img class="warning" style="float: left; margin-bottom: 25px;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>WARNING: During this adjustment procedure, operation does not stop when the door is open. To avoid injury, do not reach into the autosampler during this procedure. NOTE: It is not necessary to adjust the vertical position.</strong> <div class="ts-question">Is the sample needle correctly entering the sample vial?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Injection Port Position]]</span> <span class="btn-no">[[No->ADJUST RACK Parameter]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Injection Port Position</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/injection-port-sil.jpg" alt="Injection Port Position"></div> <h3>Examine the needle as it enters the injection port.</h3> The ADJUST INJ PORT parameter is used to adjust the needle position as it enters the injection port. If the needle is not centered over the center of the injection port, the needle may be damaged. <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>WARNING: During this adjustment procedure, operation does not stop when the door is open.To avoid injury, do not reach into the autosampler during this procedure.</strong> <div class="ts-question">Is the sample needle correctly entering the injection port?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Measuring Pump (1646 / 1571 /1673)]]</span> <span class="btn-no">[[No->Injection Port Adjustment]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">ADJUST RACK Parameter</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/WwYIR2O124Q?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left; margin-bottom:25px;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>WARNING: During this adjustment procedure, operation does not stop when the door is open. To avoid injury, do not reach into the autosampler during this procedure. NOTE: It is not necessary to adjust the vertical position.</strong> <ul> <li>← Moves the needle 0.1 mm to the left.</li> <li>→ Moves the needle 0.1 mm to the right</li> <li>. ↑ Moves the needle all the way up.</li> <li>↓ Moves the needle 6.0 mm in a downward direction.</li> <li>FUNC Moves the needle forward 0.1 mm.</li> <li>BACK Moves the needle backward 0.1 mm.</li> </ul> <ol> <li>From the Initial screen, press the VP button (x4). The Calibration Support Group screen is displayed.</li> <li>Press the FUNC key to display the Input PASSWORD screen.</li> <li>Enter the password (0,0,0,0,0) and press ENTER. The ADJUST MTP screen is displayed.</li> <li>Press FUNC until the ADJUST RACK screen is displayed.</li> <li>Press ENTER.</li> <li>Set a sample vial in position 1 of the vial rack. Do not attach a septum to the vial.</li> <li>Press ENTER. The needle moves and stops above position 1 of the sample vial rack.</li> <li>Using the keys, adjust the lateral and longitudinal position of the needle so that the needle is lowered into the center of the sample vial.</li> <li>Press ENTER. Adjustment is complete and the needle moves back to the injection port.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Measuring Pump</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/measuring-pump.jpg" alt="Measuring Pump"></div> <h3>If the measuring pump plunger becomes soiled or scratched, it can interfere with the injection precision.</h3> The measuring pump pulls the sample solution into the sample loop and pushes it into the flow path. If the measuring pump is not functioning properly, it is possible that sample is not being injected into the flow path. If no sample is injected, there will not be any peaks in the chromatogram. Inspect the plunger for scratches or debris. Scratches or foreign matter on the plunger may interfere with the seal and prohibit sample aspiration. Some of the things that may cause the plunger to be scratched are: <ul> <li>Particulate matter in the rinse solution</li> <li>Running out of rinse solution</li> <li>Worn plunger seal.</li> </ul> If the measuring pump plunger becomes soiled or scratched, it can interfere with the injection precision. <div class="ts-question">Has the measuring pump plunger been inspected recently?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Low-Pressure Valve]]</span> <span class="btn-no">[[No->Plunger Inspection and Replacement - SIL-30AC]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Injection Port Adjustment</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/3Hgefu6z8ec?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>WARNING: During this adjustment procedure, operation does not stop when the door is open.To avoid injury, do not reach into the autosampler during this procedure.</strong> <ol> <li>From the Initial screen, press the VP button four times. The Calibration Support Group screen is displayed</li> <li>Press the FUNC key to display the Input PASSWORD screen.</li> <li>Enter the password (0,0,0,0,0) and press ENTER. The ADJUST MTP screen is displayed.</li> <li>Press FUNC until the Injection Port fine adjustment screen is displayed.</li> <li>Press ENTER. The needle moves to the front center position.</li> <li>Remove the thumbscrews that secure the clear front panel. Remove the clear front panel.</li> <li>Loosen the three thumbscrews that secure the Z-mount cover. Pull the Z-mount cover forward to remove it.</li> <li>Press ENTER. The Set New Position screen is displayed.</li> <li>Using the following keys, adjust the lateral and longitudinal needle position to align with the center of the top edge of the injection port.</li> <li>Press ENTER. The needle moves up and down about 2 mm.</li> <li>Enter the set value.</li> <li>Verify that the adjustment is correct and press 1. The needle moves back to the front center position.</li> <li>Re-attach the Z-mount cover.</li> <li>Press ENTER.The fine adjustment of the needle at the injection port is complete and the needle moves back to the injection port.</li> <li>Re-attach the clear front panel and secure it with the 5 thumbscrews.</li> <li>Press 1 to start the aging process. The Injecting screen is displayed.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Plunger Seal Replacement - SIL-30AC</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/x24Do1EqZ74?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: If the pump head is pulled out or pushed in diagonally, the plunger could be broken.</strong> Remove any contamination present on the pump head and seal holder. Wipe the measuring pump head, inside of the pump head and seal holder using soft cloth or paper soaked with 2-propanol. <ol> <li>Press [CE] repeatedly to display the initial screen.</li> <li>Press [VP] repeatedly to display the Maintenance Information screen.</li> <li>Press [func] repeatedly to display the [P-SET] screen.</li> <li>Press [enter] to fully retract the plunger.</li> <li>Open the doors.</li> <li>Use the provided wrench to remove the tubing from the top and bottom of the pump head.</li> <li>Remove the tubing from port No. 2 and the center port of the low-pressure valve.</li> <li>Remove the manual prime valve.</li> <li>Use the provided Allen wrench to remove the two hex socket head bolts in the pump head.</li> <li>Gently remove the pump head.</li> <li>Carefully remove the plunger seal holder.</li> <li>Insert the flanged end of the seal installer/remover tool into the seal vertically.</li> <li>Pull out the used seal vertically. Then remove the seal from the seal tool.</li> <li>Place the new seal on the straight end of the seal installer/remover tool.</li> <li>Lower the seal into the seal holder, then remove the seal installer/remover tool.</li> <li>Reinstall the plunger seal holder.</li> <li>Reattach the pump head with the two hex socket head bolts.</li> <li>Tighten them alternately and uniformly with the provided Allen wrench.</li> <li>Then hand-tighten the manual prime valve.</li> <li>Reattach the tubing to the top and bottom of the pump.</li> <li>Close the doors.</li> </ol> <span class="btn-link">[[Plunger Inspection and Replacement - SIL-30AC->Plunger Inspection and Replacement - SIL-30AC]]</span> <span class="btn-link">[[Plunger Inspection and Replacement - SIL-30AC MP->Plunger Inspection and Replacement - SIL-30AC MP]]</span> <span class="btn-link">[[Plunger Seal Replacement - SIL-30AC MP->Plunger Seal Replacement - SIL-30AC MP]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Waste Discharge</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/waste.jpg" alt="Waste Discharge"></div> <h3>Determine whether the problem is related to the solvent delivery system.</h3> Examine the waste tubing as it enters the waste container. Is the flow of the discharge appropriate for the flow rate? <div class="ts-question">Is the mobile phase flowing?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Room Temperature (1597)]]</span> <span class="btn-no"><a href="https://www.ssi.shimadzu.com/service-support/virtual-advisor/nexera/advanced-troubleshooting/troubleshooting-the-pump/index.html">Troubleshoot the Pump</a></span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Ghost Peaks</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/ghost-peak.jpg" alt="Ghost Peaks"></div> It is possible that the magnitude of the analyte peak may hide any carryover peak. Make an injection of blank sample solvent to verify whether there is a carryover peak eluting at the same time as the analyte peak. If the extra peak has a different retention time from the analyte peak, it is most likely a system peak that could be caused from a variety of system contamination sources. Some of these sources include: <ul> <li>Injector contamination</li> <li>Detector contamination</li> <li>Column contamination</li> <li>Gradient issues</li> </ul> <div class="ts-question">Are extra peaks or ghost peaks appearing in the chromatogram?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Rinse Solution 1 (1612)]]</span> <span class="btn-no">[[No->Relative Standard Deviation]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Autosampler (SIL) Plumbing</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/plumbing.jpg" alt="Autosampler (SIL) Plumbing"></div> <ol> <li>Connect the autosampler inlet tubing to the pump outlet. The inlet tubing, marked with a blue band, emerges from the right side of the autosampler and is connected to port 6 of the HPV.</li> <li>Pass the autosampler outlet tubing through the tubing port of the column oven and connect it to the column inlet. The outlet tubing emerges from the right side of the autosampler and is connected to port 5 of the HPV.</li> <li>Connect the SS rinse inlet tubing on top of the autosampler to the degasser outlet using the fittings supplied with the degasser. The rinse phase inlet tubing is connected to port 4 on the low-pressure valve.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Column Performance</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/column-param.jpg" alt="Column Performance"></div> It is necessary to use a column logbook to record the performance of the column. Record the initial column parameters such as pressure, analyte retention time, Resolution (R), Plate count (N) and Tailing Factor (T). By periodically recording this information, it is possible to determine when a column needs to be flushed, regenerated or retired. <div class="ts-question">Has the column performance deteriorated?</div> <div class="buttons"><span class="btn-yes">[[Yes->Column Regeneration (1311)]]</span> <span class="btn-no">[[No->Tubing Connections]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Column Regeneration</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/column.jpg" alt="Column Regeneration"></div> <strong>Reversed-Phase Column Regeneration</strong> <ol> <li>Disconnect the column and reconnect it to the pump in the reverse direction. Do not connect it to the detector.</li> <li>Flush out the salts and or buffers with 25 mL of mobile phase (without buffer) at 1mL/min.</li> <li>Flush the column with 25 mL of HPLC grade water.</li> <li>Flush the column with 25 mL of isopropanol.</li> <li>Flush the column with 25 mL of methylene chloride.</li> <li>Flush the column with 25 mL of hexane.</li> <li>Remove the hexane by flushing again with 25 mL of methylene chloride.</li> <li>Remove the methylene chloride by flushing again with 25 mL of isopropanol.</li> <li>Reconnect the column in the proper direction and flush the column with the mobile phase (without buffer), then reintroduce the buffer.</li> <li>Equilibrate the column with 25-50 mL of mobile phase.</li> <li>Inject a standard solution to re-evaluate column performance.</li> </ol> <strong>Normal-Phase Column Regeneration</strong> <ol> <li>Disconnect the column and reconnect it to the pump in the reverse direction. Do not connect it to the detector.</li> <li>Flush the column with 50 mL of 50:50 methanol:chloroform.</li> <li>Flush the column with 50 mL of ethyl acetate.</li> <li>Reconnect the column in the proper direction.</li> <li>Equilibrate the column with 50 mL of mobile phase.</li> <li>Inject a standard solution to re-evaluate column performance.</li> </ol> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>NOTE: In some cases, dimethyl-formamide might be a better choice of 'cleaning' solvent than the methylene chloride and hexane. Always refer to the column vendor for the recommended washing technique for that column type.</strong> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Tubing Connections</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/flow.jpg" alt="Tubing Connections"></div> <h3>Improper tubing connections may be allowing dead volume to be incorporated in the flow path.</h3> The tubing into and out of the autosampler may cause some confusion. The inlet tubing brings mobile phase from the pump or mixer into port 6 of the high-pressure valve.This inlet tubing IS marked with a blue band. The outlet tubing carries the mobile phase and sample from port 5 of the high-pressure valve to the column inlet. This outlet tubing IS NOT marked with a blue band. Improper tubing connections may be allowing dead volume to be incorporated in the flow path. <div class="ts-question">Are the autosampler tubing connections correct?</div> <div class="buttons"><span class="btn-yes">[[Yes->Flow Line Leaks 1 (1587)]]</span> <span class="btn-no">[[No->Tubing - Flow Line Connections (1954)]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Flow Line Leaks</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-pump/flow-lines.jpg" alt="Flow Line Leaks"></div> A leak in the flow line before the detector flow cell could explain the absence of sample peaks. Visually inspect the flow line from needle to flow cell looking for signs of a leak: <ul> <li>Puddles</li> <li>Drips</li> <li>Crystallization of buffer</li> </ul> <div class="ts-question">Are the flow line connections leaking?</div><div class="buttons"> <span class="btn-yes">[[Yes->Tubing - Flow Line Connections (1954)]]</span> <span class="btn-no">[[No->Clogged Flow Lines (1590)]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Tubing - Flow Line Connections</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-pump/tubing-flow-line-connections.jpg" alt="Tubing - Flow Line Connections"></div> <ol> <li>Place a male nut and ferrule on the tubing.</li> <li>Insert the end of the tubing with the ferrule completely into the appropriate fitting.</li> <li>Push the tubing into the fitting until it butts up against the end of the fitting (tubing stop position).</li> <li>While making sure the tubing remains in the tubing stop position, securely tighten the male nut. The ferrule is secured to the tubing.</li> </ol> <img class="warning" style="float: left; " src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>NOTE: It is important to apply a significant amount of pressure to the tubing while tightening the male nut.</strong> <span class="btn-link">[[Cutting Resin ->Cutting Resin]]</span> <span class="btn-link">[[Cutting SS ->Cutting SS]]</span> <span class="btn-link">[[Tighten SS ->Tighten SS]]</span> <div class="buttons"><span class="btn-back"><<back "Back">></span></div> </div>

<div class="container"><h1 class="title">Tubing - Cutting Resin</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-pump/tubing-cutting-resin.jpg" alt="Tubing - Cutting Resin"></div> When using PEEK tubing or equivalent resin tubing, use a straight edge razor knife to cut the tubing. To avoid the creation of void volumes, verify that the cut edges are clean right-angled cuts. <span class="btn-link">[[Flow Line Connections ->Tubing - Flow Line Connections (1954)]]</span> <span class="btn-link">[[Cutting SS ->Cutting SS]]</span> <span class="btn-link">[[Tighten SS ->Tighten SS]]</span> <div class="buttons"><span class="btn-back"><<back "Back">></span></div> </div>

<div class="container"><h1 class="title">Tubing - Cutting SS</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-pump/tubing-cutting-ss.jpg" alt="Tubing - Cutting SS"></div> When using HPLC stainless steel tubing, it is best to use precut stainless steel tubing or employ a cutting technique that produces a smooth and even surface to the cut edge. If the cut edge of the tubing is not smooth and square, a void volume is created in the connection. <span class="btn-link">[[Cutting Resin ->Cutting Resin]]</span> <span class="btn-link">[[Flow Line Connections ->Tubing - Flow Line Connections (1954)]]</span> <span class="btn-link">[[Tighten SS ->Tighten SS]]</span> <div class="buttons"><span class="btn-back"><<back "Back">></span></div> </div>

<div class="container"><h1 class="title">Tubing - Tighten SS</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-pump/tubing-tighten-ss.jpg" alt="Tubing - Tighten SS"></div> Verify that the stainless steel unions are securely connected. Use two wrenches to tighten stainless steel couplings. <span class="btn-link">[[Flow Line Connections ->Tubing - Flow Line Connections (1954)]]</span> <span class="btn-link">[[Cutting SS ->Cutting SS]]</span> <span class="btn-link">[[Cutting Resin ->Cutting Resin]]</span> <div class="buttons"><span class="btn-back"><<back "Back">></span></div> </div>

<div class="container"><h1 class="title">Clogged Flow Lines</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/flow-sil.jpg" alt="Clogged Flow Lines"></div> <h3>The flow lines in the injection path may be clogged or partially blocked and may need to be replaced.</h3> The autosampler flow line is detailed above. Ensure that the flow lines are plumbed correctly. Please examine these flow lines and connections for blockage or leaks. The flow lines in the injection path may be clogged or partially blocked and may need to be replaced. <div class="ts-question">Are the flow lines in the injection flow path clogged or partially blocked?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Tubing Replacement (1545 / 1591)]]</span> <span class="btn-no"><a href="https://www.ssi.shimadzu.com/forms/tech-support/index.html?product_category=VirtualAdvisor&product_name=Nexera&page=webform_start">Contact Shimadzu</a></span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Room Temperature</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/column-temp.jpg" alt="Room Temperature"></div> <ul> <li>Upward drifting baselines usually result from an inadequately equilibrated column. Allow the column to equilibrate until the baseline stabilizes.</li> <li>Downward drifting baselines are often caused by not allowing the lamp to heat sufficiently. Allow the lamp to heat for 30 minutes before examining the baseline.</li> <li>A meandering baseline may be related to the cyclical temperature fluctuation of the column heating device.Use of a column oven can reduce the magnitude of this temperature cycling.</li> <li>A wandering baseline is often related to ambient temperature fluctuations or contaminants eluting from a column. Thermally isolate the column and other system components that can be affected by temperature changes, and wash the column thoroughly.</li> </ul> <div class="ts-question">Is the room temperature fluctuating?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Column Insulation (1598)]]</span> <span class="btn-no">[[No->Column Performance (1599)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Column Insulation</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/column-temp.jpg" alt="Column Insulation"></div> <h3>A column oven can be used to isolate the column and flow lines from ambient temperature fluctuations.</h3> Maintaining the column temperature at a constant level increases analysis reproducibility and separation performance. There are three different types of thermal insulation devices available for the LC column. <ul> <li>CTO-30A and the CTO-30AS with the SIL-30AC MP Column Oven - Uses a heating block to control the temperature of the LC column.</li> <li>CTO-20A/20AC Column Oven - Uses an air-circulated thermostatic chamber to maintain the temperature of the LC column and flow lines.</li> <li>Jacket-type Devices - Uses plumbing insulation or heat tape to a lesser degree of effectiveness</li> </ul> <div class="ts-question">Does the system include a column oven?</div> <div class="buttons"> <span class="btn-yes"><a href="https://www.ssi.shimadzu.com/service-support/virtual-advisor/nexera/advanced-troubleshooting/troubleshooting-the-columnoven/index.html">Troubleshoot the Column Oven</a></span> <span class="btn-no">[[No->Column Oven (1608)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Column Oven</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/cto-front.jpg" alt="Column Oven"></div> <h3>A column oven can be used to isolate the column and flow lines from ambient temperature fluctuations.</h3> The Shimadzu column ovens (CTO-30A/30AS) use a heating block to control the temperature of the LC column to improve retention time reproducibility and separation performance. The Shimadzu column ovens (CTO-20A/20AC) are air-circulated thermostatic chambers designed to improve retention time reproducibility and separation performance by maintaining the temperature of the LC column and flow lines. In addition to the column, the oven can also house manual injectors, solvent mixers, column switching valves and other flow line accessories. Installing these items into the column oven isolates them from the influence of ambient temperature fluctuations and allows more stable analysis. <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Rinse Solution</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/bottle.jpg" alt="Rinse Solution"></div> <h3>Examine the rinse solution reservoir.</h3> Although the injection process may still function if the rinse solution runs dry, the reproducibility will suffer greatly. When the rinse solution is used up, air may be drawn into the injection tubing. Air in the injection tubing interferes with the reproducibility of the injection process. In such cases, the autosampler must be manually purged. Examine the rinse solution reservoir. <div class="ts-question">Is the volume of rinse solution adequate for the analysis?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Rinse Volume 1 (1615)]]</span> <span class="btn-no">[[No->MANUAL PURGE (10479)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Relative Standard Deviation</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/rsd.jpg" alt="Relative Standard Deviation"></div> Determine whether the peak areas are consistent from one injection to another using the following procedure. Make 3-5 injections from a single vial of a known standard solution. Most computer based chromatography software can automatically calculate %RSD. Consult the software documentation or use the calculations above to calculate the mean (average) area, the standard deviation and the %RSD. Passing %RSD is typically less than 1%. <div class="ts-question">Has the reproducibility of known peaks changed?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Rinse Solution 2 (1657)]]</span> <span class="btn-no">[[No->Baseline Stability]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Rinse Volume</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/rinse-port.jpg" alt="Rinse Volume"></div> Depending on the viscosity of the sample solution, sample carryover from one injection to another can be a problem in HPLC analysis. The most common way of reducing this affect is by rinsing the sampling needle between samples. The Shimadzu Prominence series HPLC autosampler has a number of improvements that greatly minimize this affect, such as the addition of a specially coated needle and a change in the injection port design. It is also possible to add another rinse pump to the SIL-20 autosamplers to add the capability of an active rinse of the outer surface of the needle. Shimadzu autosamplers provide for a dip rinse of the outer surface of the needle. Now, with the addition of the Additional Rinse Pump it is possible to use two different rinse solvents and perform both the dip rinse and the active rinse. <div class="ts-question">Is the RINSE VOLUME parameter adequate for the sample type?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Injection Port Purge]]</span> <span class="btn-no">[[No->RINSE VOLUME Parameter]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">MANUAL PURGE</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/l_BJzmGimOk?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <strong>It is a good practice to follow all manual purges with an automatic purge.</strong> <ol> <li>From the Initial screen, press the FUNC key twice. The CONTROL screen is displayed.</li> <li>Press ENTER. The Clear SMPTBL (clear sample table) screen is displayed.</li> <li>Press the FUNC key four times. The MANUAL PURGE screen is displayed.</li> <li>Insert the manual syringe into the tubing that is connected to port 3 of the low-pressure valve.</li> <li>Press 1 on the numeric keypad.</li> <li>Draw in the rinse solution with the manual syringe.</li> <li>Press 0 on the numeric keypad. The MANUAL PURGE process is finalized.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Injection Port Purge</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/mobile-phase.jpg" alt="Injection Port Purge"></div> <h3>When changing mobile phase, it is important to purge the injection flow path.</h3> A sufficient supply of mobile phase is required to perform LC measurement. Before the degasser can be turned ON, all of the flow lines must be filled with solvent by manually purging the pump. When changing mobile phase, it is important to purge the injection flow path. <div class="ts-question">Has the injection port been purged to remove the old mobile phase?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Rinse Purge]]</span> <span class="btn-no">[[No->MANUAL PURGE (10479)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">RINSE VOLUME Parameter</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/035kyF0K6Ag?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>NOTE: If the rinse volume is set to [0], analysis time is reduced by omitting the purge operation for the measurement flow lines after sample injection.</strong> <ol> <li>From the RINSE MODE screen, press the FUNC key twice. The RINSE VOLUME screen is displayed.</li> <li>Enter the numeric value for the parameter using the numeric keypad and press the ENTER key.</li> </ol> <span class="btn-link">[[RINSE MODE Parameter->RINSE MODE Parameter]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Rinse Purge</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/bottle.jpg" alt="Rinse Purge"></div> <h3>When changing the rinse phase solution, it is important to purge the rinse flow lines.</h3> Although the injection process may still function if the rinse solution runs dry, the reproducibility will suffer greatly. When the rinse solution is used up, air may be drawn into the injection tubing. Air in the injection tubing interferes with the reproducibility of the injection process. In such cases, the autosampler must be manually purged. When changing the rinse phase solution, it is important to purge the rinse flow lines. <div class="ts-question">Have the rinse flow lines been purged to remove the old rinse solution?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Needle Seal 1 (1643)]]</span> <span class="btn-no">[[No->MANUAL PURGE (10479)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Needle Seal</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/needle-seal.jpg" alt="Needle Seal"></div> <h3>The needle seal is a consumable part and needs to be replaced periodically.</h3> The needle seal provides a leak-free injection port for introducing the sample into the HPLC system. The needle seal is a consumable item for the Prominence series of autosamplers. The needle seal should be replaced after about 40,000 injections. Use the NDL SEAL USED function (Maintenance Information group) to monitor the needle seal usage. Use the NDL Seal alert function (Calibration Support group) to set the level of needle seal usage that will cause an alarm to sound. <div class="ts-question">Has the needle seal been replaced recently?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Measuring Pump (1646 / 1571 /1673)]]</span> <span class="btn-no">[[No->Needle Seal Replacement - SIL-30AC]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Needle Seal Replacement - SIL-30AC</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/XN8uHCNKLmI?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: If the needle seal cannot be removed from the housing, do not attempt to pull it out by force. Replace both the needle seal and the housing at the same time. Attempting to pull it out by force may result in injury. CAUTION: When inserting the needle seal into the housing, hold the needle seal using a piece of soft cloth or paper, being careful not to allow dust to enter the hole. The entry of dust may create small gaps or voids that could lead to carryover or leaks.</strong> <ol> <li>Press [ZHOME Key] on the initial screen. The [ZHOME] screen is displayed.</li> <li>Press [enter] to move the needle up fully and to the center of the instrument.</li> <li>Turn OFF the power switch and disconnect the power cord.</li> <li>Open the doors.</li> <li>Remove the knurled thumbscrews (5) from the cooler cover.</li> <li>Pull on the cooler cover and remove it.</li> <li>Loosen the cap using the supplied needle seal XR installer / remover tool, and remove it.</li> <li>Use tweezers to remove the housing and the needle seal from the stator.</li> <li>Remove the needle seal from the housing. See CAUTION below.</li> <li>Insert the housing into the stator.</li> <li>Insert a new needle seal into the housing with the tapered hole on the needle seal facing upward. See CAUTION above.</li> <li>Finger-tighten the cap on the stator.</li> <li>Use the needle seal XR installer / remover tool to tighten the cap an additional 180°.</li> <li>Reinstall the sample cooler front panel.</li> <li>Secure the cooler panel with the knurled thumbscrews (5).</li> <li>Close the doors.</li> </ol> <span class="btn-link">[[Needle Seal Replacement - SIL-30AC MP->Needle Seal Replacement - SIL-30AC MP]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

Combined two "Measuring the Pump" https://nexerava.ssi.shimadzu.com/node/1646 https://nexerava.ssi.shimadzu.com/node/1571 https://nexerava.ssi.shimadzu.com/node/1673 In regard to above, I linked to Plunger Inspection and Replacement - SIL-30AC" instead of "Plunger Seal Replacement - SIL-30AC" since it is more comprehensive https://nexerava.ssi.shimadzu.com/node/668 https://nexerava.ssi.shimadzu.com/node/670 Combined "Clogged Flow Lines" https://nexerava.ssi.shimadzu.com/node/1590 https://nexerava.ssi.shimadzu.com/node/1700 Combined "Tubing Replacement" https://nexerava.ssi.shimadzu.com/node/1591 https://nexerava.ssi.shimadzu.com/node/1545 Combined "Room Temperature" ???????????? https://nexerava.ssi.shimadzu.com/node/1597 https://nexerava.ssi.shimadzu.com/node/1688 Combined "Column Insulation" ??????????????? https://nexerava.ssi.shimadzu.com/node/1689 https://nexerava.ssi.shimadzu.com/node/1598 https://nexerava.ssi.shimadzu.com/node/1670

<div class="container"><h1 class="title">Low-Pressure Valve</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/low-pressure-valve.jpg" alt="Low-Pressure Valve"></div> <h3>The low-pressure valve rotor and stator are consumable parts and need to be replaced periodically.</h3> The low-pressure valve (LPV) is responsible for the flow control of the rinse solution. If the LPV is not functioning properly, rinse solution may not reach the injection port and needle. The LPV rotor should be replaced after about 60,000 injections. Since the rinse solution removes any residual sample left in the injection areas, extra peaks may become evident if the LPV malfunctions. The low-pressure valve rotor and stator are consumable parts and need to be replaced periodically. <div class="ts-question">Have the low-pressure valve rotor and stator been replaced recently?</div> <div class="buttons"> <span class="btn-yes"><a href="https://www.ssi.shimadzu.com/forms/tech-support/index.html?product_category=VirtualAdvisor&product_name=Nexera&page=webform_start">Contact Shimadzu</a></span> <span class="btn-no">[[No->LPV Rotor and Stator Replacement - SIL-30AC]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Plunger Inspection and Replacement - SIL-30AC</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/4AwNCxeblQM?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: When sliding the plunger tool onto the plunger, be careful not to damage the plunger. </strong> The measurement accuracy may be reduced if there is any seal residue or other matter on the plunger. Screw the bolts in alternately 90°. At the end, tighten the bolts securely with the Allen wrench. <ol> <li>Press [CE] repeatedly to display the initial screen.</li> <li>Press [VP] repeatedly to display the Maintenance Information screen.</li> <li>Press [func] repeatedly to display the [P-SET] screen.</li> <li>Press [enter] to fully retract the plunger.</li> <li>Open the doors.</li> <li>Use the provided wrench to remove the tubing from the top and bottom of the pump head.</li> <li>Remove the tubing from port No. 2 and the center port of the low-pressure valve.</li> <li>Remove the manual prime valve.</li> <li>Use the provided Allen wrench to remove the two hex socket head bolts in the pump head.</li> <li>Gently remove the pump head.</li> <li>Carefully remove the plunger seal holder.</li> <li>Use the plunger tool to remove the plunger ASSY.</li> <li>Use a soft cloth soaked in 2-propanol to wipe the inside of the pump head and seal holder.</li> <li>Inspect the plunger for visible nicks or scratches. Replace the plunger ASSY if necessary.</li> <li>Hand-tighten the plunger ASSY into the pump body.</li> <li>Use the plunger tool to thoroughly tighten the plunger ASSY.</li> <li>Reinstall the plunger seal holder.</li> <li>Reattach the pump head with the two hex socket head bolts.</li> <li>Tighten them alternately and uniformly with the provided Allen wrench.</li> <li>Then hand-tighten the manual prime valve.</li> <li>Reattach the tubing to the top and bottom of the pump.</li> <li>Close the doors.</li> </ol> <span class="btn-link">[[Plunger Seal Replacement - SIL-30AC->Plunger Seal Replacement - SIL-30AC]]</span> <span class="btn-link">[[Plunger Inspection and Replacement - SIL-30AC MP->Plunger Inspection and Replacement - SIL-30AC MP]]</span> <span class="btn-link">[[Plunger Seal Replacement - SIL-30AC MP->Plunger Seal Replacement - SIL-30AC MP]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Plunger Inspection and Replacement - SIL-30AC MP</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/OSE5xi8rCn8?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: When sliding the plunger tool onto the plunger, be careful not to damage the plunger. </strong> The measurement accuracy may be reduced if there is any seal residue or other matter on the plunger. Screw the bolts in alternately 90°. At the end, tighten the bolts securely with the Allen wrench. <ol> <li>Press [CE] repeatedly to display the initial screen.</li> <li>Press [VP] repeatedly to display the Maintenance Information screen.</li> <li>Press [func] repeatedly to display the [P-SET] screen.</li> <li>Press [enter] to fully retract the plunger.</li> <li>Open the right door.</li> <li>Use the provided wrench to remove the tubing from the top and bottom of the pump head.</li> <li>Remove the tubing from port No. 2 and the center port of the low-pressure valve.</li> <li>Remove the manual prime valve.</li> <li>Use the provided Allen wrench to remove the two hex socket head bolts in the pump head.</li> <li>Gently remove the pump head.</li> <li>Carefully remove the plunger seal holder.</li> <li>Use the plunger tool to remove the plunger ASSY.</li> <li>Use a soft cloth soaked in 2-propanol to wipe the inside of the pump head and seal holder.</li> <li>Inspect the plunger for visible nicks or scratches. Replace the plunger ASSY if necessary.</li> <li>Hand-tighten the plunger ASSY into the pump body.</li> <li>Use the plunger tool to thoroughly tighten the plunger ASSY.</li> <li>Reinstall the plunger seal holder.</li> <li>Reattach the pump head with the two hex socket head bolts.</li> <li>Tighten them alternately and uniformly with the provided Allen wrench.</li> <li>Then hand-tighten the manual prime valve.</li> <li>Reattach the tubing to the top and bottom of the pump.</li> <li>Close the right door.</li> </ol> <span class="btn-link">[[Plunger Seal Replacement - SIL-30AC->Plunger Seal Replacement - SIL-30AC]]</span> <span class="btn-link">[[Plunger Inspection and Replacement - SIL-30AC->Plunger Inspection and Replacement - SIL-30AC]]</span> <span class="btn-link">[[Plunger Seal Replacement - SIL-30AC MP->Plunger Seal Replacement - SIL-30AC MP]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Plunger Seal Replacement - SIL-30AC MP</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/SPtS885kZnw?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: If the pump head is pulled out or pushed in diagonally, the plunger could be broken.</strong> <ol> <li>Press [ZHOME Key] on the initial screen. The [ZHOME] screen is displayed.</li> <li>Press [enter] to move the needle up fully and to the center of the instrument.</li> <li>Turn OFF the power switch and disconnect the power cord.</li> <li>Pull on the L-shaped faceplate to remove it.</li> <li>Open the key panel.</li> <li>Remove any sample racks.</li> <li>Open the rack door.</li> <li>Open the right and left doors.</li> <li>Use the provided wrench to remove the tubing from the top and bottom of the pump head.</li> <li>Remove the tubing from port No. 2 and the center port of the low-pressure valve.</li> <li>Remove the manual prime valve.</li> <li>Use the provided Allen wrench to remove the two hex socket head bolts in the pump head.</li> <li>Gently remove the pump head.</li> <li>Carefully remove the plunger seal holder.</li> <li>Insert the flanged end of the seal installer/remover tool into the seal vertically.</li> <li>Pull out the used seal vertically. Then remove the seal from the seal tool.</li> <li>Place the new seal on the straight end of the seal installer/remover tool.</li> <li>Lower the seal into the seal holder, then remove the seal installer/remover tool.</li> <li>Reinstall the plunger seal holder.</li> <li>Reattach the pump head and tighten the two hex socket head bolts alternately and uniformly with the provided Allen wrench.</li> <li>Then hand-tighten the manual prime valve.</li> <li>Reattach the tubing to the top and bottom of the pump.</li> <li>Close the doors and install the sample racks.</li> </ol> <span class="btn-link">[[Plunger Seal Replacement - SIL-30AC->Plunger Seal Replacement - SIL-30AC]]</span> <span class="btn-link">[[Plunger Inspection and Replacement - SIL-30AC->Plunger Inspection and Replacement - SIL-30AC]]</span> <span class="btn-link">[[Plunger Inspection and Replacement - SIL-30AC MP->Plunger Inspection and Replacement - SIL-30AC MP]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">RINSE MODE Parameter</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/18SFhx0YEM8?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> The RINSE MODE parameter selects the needle rinsing method. <ul> <li>0 - No Rinsing</li> <li>1 - Rinsing before sample aspiration</li> <li>2 - Rinsing after sample aspiration</li> <li>3 - Rinsing before and after sample aspiration (default)</li> </ul> <ol> <li>From the Initial screen, press the FUNC key. The Parameter setting screen is displayed.</li> <li>Press the ENTER key and then press the FUNC key twice. The RINSE MODE parameter screen is displayed.</li> <li>Enter the rinse mode number with the numeric keypad and press the ENTER key.</li> </ol> <span class="btn-link">[[RINSE VOLUME Parameter->RINSE VOLUME Parameter]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Needle Seal Replacement - SIL-30AC MP</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/fNWy10xPRuc?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: If the needle seal cannot be removed from the housing, do not attempt to pull it out by force. Replace both the needle seal and the housing at the same time. Attempting to pull it out by force may result in injury. CAUTION: When inserting the needle seal into the housing, hold the needle seal using a piece of soft cloth or paper, being careful not to allow dust to enter the hole. The entry of dust may create small gaps or voids that could lead to carryover or leaks.</strong> <ol> <li>Press [ZHOME Key] on the initial screen. The [ZHOME] screen is displayed.</li> <li>Press [enter] to move the needle up fully and to the center of the instrument.</li> <li>Turn OFF the power switch and disconnect the power cord.</li> <li>Pull on the L-shaped faceplate to remove it.</li> <li>Open the key panel.</li> <li>Remove any sample racks.</li> <li>Open the rack door.</li> <li>Open the right and left doors.</li> <li>Remove the knurled thumbscrews (6) from the cooler cover.</li> <li>Pull on the cooler cover and remove it.</li> <li>Loosen the cap using the supplied needle seal XR installer / remover tool, and remove it.</li> <li>Remove the housing and the needle seal from the stator.</li> <li>Remove the needle seal from the housing. See CAUTION message.</li> <li>Insert the housing into the stator.</li> <li>Insert a new needle seal into the housing with the tapered hole facing upward. See CAUTION message.</li> <li>Replace and finger-tighten the cap on the stator.</li> <li>Use the needle seal tool to tighten the cap an additional 180°.</li> <li>Reattach the sample cooler cover and close all of the doors.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Baseline Stability</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/baseline-stability.jpg" alt="Baseline Stability"></div> Make a sample injection to calculate the signal-to-noise ratio and determine if it is acceptable for your application. <ul> <li>All baselines have an inherent amount of noise. Baseline noise is only a problem if the amount of noise is so excessive as to interfere with integration of the analyte of interest. Data systems often have an auto scale function that can make a normal baseline appear to be very noisy.</li> <li>The signal to noise ratio is a measure of the Peak height with respect to the Noise height. This ratio can be increased by decreasing the noise or by increasing the peak (signal).</li> <li>The sensitivity of the method and the scale of the chromatogram output must be determined before determining whether or not the noise is excessive.</li> </ul> <img class="warning" style="float: left; margin-bottom:25px;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>NOTE: Calculating Signal-to-Noise Ratio. The signal-to-noise ratio is calculated using the peak height of the noise and the peak height of the sample peak. EXAMPLE: Noise height 2mV - Sample height 10mV - The signal-to-noise ratio is 5. Typically, a signal-to-noise ratio of 3 is required for detection, and 10 for quantitation.</strong> <div class="ts-question">Is the baseline drifting?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Detector Function 2 (1684)]]</span> <span class="btn-no">[[No->High-pressure Valve Pressure Fluctuation]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">High-pressure Valve Pressure Fluctuation</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/flow-path-ready.jpg" alt="High-pressure Valve Pressure Fluctuation"></div> <h3>Monitor the pressure on the pump display through a complete injection sequence.</h3> The high-pressure valve directs the flow changes during the injection process. Monitor the pressure display on the solvent delivery unit to determine whether the pressure fluctuates widely during the injection process. Monitor the pressure on the pump display through a complete injection sequence. <div class="ts-question">Does the pressure fluctuate during switching of the high-pressure valve?</div> <div class="buttons"> <span class="btn-yes">[[Yes->High-Pressure Valve Rotation]]</span> <span class="btn-no">[[No->Leak in the Autosampler]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Detector Function</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/detector-saturation.jpg" alt="Detector Function"></div> It is possible that a problem with the detector may manifest itself as a problem with the autosampler. <span class="btn-link"><a href="https://www.ssi.shimadzu.com/service-support/virtual-advisor/nexera/advanced-troubleshooting/troubleshooting-pda-detector/index.html">Troubleshoot the Detector</a></span> <div class="ts-question">Is the detector functioning properly?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Bubbles in the Flow Cell (1686)]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Bubbles in the Flow Cell</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/sawtooth.jpg" alt="Bubbles in the Flow Cell"></div> <h3>Purging the flow lines helps to remove any trapped air bubbles.</h3> Generally, the more dissolved air that is removed from the mobile phase the better the entire HPLC system will perform. However, when mobile phase does contain dissolved gas, the gas can escape from solution to form a small bubble in the flow line. These small bubbles can attach to any dirt particles that may be in the flow cell. As more gas escapes from solution, the bubble becomes larger causing a disturbance to the baseline. When the bubble gets large enough to flow out with the mobile phase, the baseline returns to zero. This phenomenon usually causes a very regular sinusoidal (sawtooth) shape to the baseline. It is also possible that the bubbles will manifest themselves as spiking vertical lines in the baseline. <div class="ts-question">Have the flow lines been purged?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Room Temperature (1688)]]</span> <span class="btn-no">[[No->MANUAL PURGE (10479)]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">High-Pressure Valve Rotation</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/flow-path-ready.jpg" alt="High-Pressure Valve Rotation"></div> <h3>Listen to the high-pressure valve through a complete injection sequence.</h3> During the injection process, the high-pressure valve rotates. The valve makes a noise during the rotation. Make an injection and listen for the appropriate noise. Listen to the high-pressure valve through a complete injection sequence. <div class="ts-question">Does it sound like the high-pressure valve is operating properly?</div> <div class="buttons"> <span class="btn-yes">[[Yes->HPV Clogging]]</span> <span class="btn-no"><a href="https://www.ssi.shimadzu.com/forms/tech-support/index.html?product_category=VirtualAdvisor&product_name=Nexera&page=webform_start">Contact Shimadzu</a></span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Leak in the Autosampler</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/leaks.jpg" alt="Leak in the Autosampler"></div> <h3>There are a number of places that leaks can occur in the autosampler.</h3> It may also be necessary to adjust the sensitivity of the leak sensor. Some of the areas that need to be inspected for leaks are listed below. <ul> <li>Flow line connections</li> <li>Sample loop connections</li> <li>High-pressure valve and connections</li> <li>Low-pressure valve and connections</li> <li>Measuring pump and connections</li> </ul> <div class="ts-question">Is there a leak in the autosampler flow path?</div> <span class="btn-link">[[Leak Sensor Adjustment->Leak Sensor Adjustment]]</span> <div class="buttons"> <span class="btn-yes">[[Yes->Sample Loop Replacement - SIL-30AC]]</span> <span class="btn-no"><a href="https://www.ssi.shimadzu.com/forms/tech-support/index.html?product_category=VirtualAdvisor&product_name=Nexera&page=webform_start">Contact Shimadzu</a></span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">HPV Clogging</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/hpv.jpg" alt="HPV Clogging"></div> <h3>It may be necessary to disassemble the high-pressure valve and clean the rotor and stator.</h3> The rotor and stator of the high-pressure valve slide against each other to direct the flow of mobile phase and sample through the injection process to the column. The groove created between the rotor and stator is very small and can become clogged easily. There are a number of factors that may cause a clog in the high-pressure valve. <ul> <li>Precipitation or undisolved buffer in the mobile phase</li> <li>Precipitation or undisolved sample</li> <li>Particles of the injection port septa</li> <li>Worn rotor particles</li> </ul> <div class="ts-question">Is the high-pressure valve clogged?</div> <div class="buttons"> <span class="btn-yes">[[Yes->HPV Rotor and Stator Removal - SIL-30AC]]</span> <span class="btn-no">[[No->High-Pressure Valve Rotor and Stator]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Sample Loop Replacement - SIL-30AC</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/P3SgA8UGBUI?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <ol> <li>Press [ZHOME Key] on the initial screen. The [ZHOME] screen is displayed.</li> <li>Press [enter] to move the needle up fully and to the center of the instrument.</li> <li>Turn OFF the power switch and disconnect the power cord.</li> <li>Open the doors.</li> <li>Remove the knurled thumbscrews (5) from the cooler cover.</li> <li>Pull on the cooler cover and remove it.</li> <li>Remove the male nut on the sample loop at port 1 of the high-pressure valve.</li> <li>Loosen the three screws on the Z mount cover and remove the Z mount cover.</li> <li>Using a wrench, unscrew and remove the male nut on the other end of the sample loop (opposite the needle).</li> <li>Remove the sample loop from the hooks (2 places) on the back of the front upper section of the plastic cover inside the instrument.</li> <li>Remove the sample loop from the hooks on the left side of the interior of the autosampler.</li> <li>Remove the sample loop from the square loop at the back of the Z mount, then remove the sample.</li> <li>Pass the new sample loop through the square loop at the back of the Z mount.</li> <li>Attach a male nut and a ferrule to the sample loop, and secure to the joint on the needle side using a wrench.</li> <li>Attach the sample loop to the hooks on the left side of the interior of the autosampler.</li> <li>Attach the sample loop to the hooks (2 places) on the back of the front upper section of the instrument.</li> <li>Secure the sample loop to port 1 of the high-pressure valve.</li> <li>Replace the Z mount cover, and tighten with its screws.</li> <li>Replace the sample cooler front panel and close the doors.</li> </ol> <span class="btn-link">[[Sample Loop Replacement - SIL-30AC MP->Sample Loop Replacement - SIL-30AC MP]]</span> <span class="btn-link">[[LPV Rotor and Stator Replacement - SIL-30AC->LPV Rotor and Stator Replacement - SIL-30AC]]</span> <span class="btn-link">[[LPV Rotor and Stator Replacement - SIL-30AC MP->LPV Rotor and Stator Replacement - SIL-30AC MP]]</span> <span class="btn-link">[[HPV Rotor and Stator Removal - SIL-30AC->HPV Rotor and Stator Removal - SIL-30AC]]</span> <span class="btn-link">[[HPV Rotor and Stator Replacement - SIL-30AC->HPV Rotor and Stator Replacement - SIL-30AC]]</span> <span class="btn-link">[[HPV Rotor and Stator Removal - SIL-30AC MP->HPV Rotor and Stator Removal - SIL-30AC MP]]</span> <span class="btn-link">[[HPV Rotor and Stator Replacement - SIL-30AC MP->HPV Rotor and Stator Replacement - SIL-30AC MP]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Leak Sensor Adjustment</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/cto-leak-sensor-replacement.jpg" alt="Leak Sensor Adjustment"></div> The leak sensor in the column oven monitors and detects the presence of organic vapor. The sensitivity of the leak sensor can be adjusted to prevent false errors or to ensure the detection of a true leak. Follow the procedure below to check the leak sensor threshold. <ol> <li>From the Initial screen press VP until the CALIBRATION screen is displayed.</li> <li>Press FUNC to display the Input PASSWORD screen.</li> <li>Using the numeric keypad, press 0, 0, 0, 0, 0, then ENTER to enter the Calibration Support Group.</li> <li>Press FUNC until the LEAK THR screen is displayed.</li> </ol> <ul> <li>The leak sensor threshold setting range is between 7000 and 40000.</li> <li>It may be necessary to replace the leak sensor if the threshold is set to the maximum setting and is still not detecting leaks.</li> </ul> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">LPV Rotor and Stator Replacement - SIL-30AC</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/VID4e2CLU0Y?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <ol> <li>Turn OFF the power switch and lower the rinse solution container to a position lower than the low-pressure valve.</li> <li>Open the doors.</li> <li>Use a wrench to remove all of the flow lines from the housing.</li> <li>Use the M4 Allen wrench to remove the three hex socket head bolts.</li> <li>Carefully, remove the stator ASSY (housing, packing, and stator) and the ring. The pieces are not connected.</li> <li>Grip the rotor securely, and pull it out of the instrument.</li> <li>Inspect the sliding surfaces of the rotor and the stator for visible nicks or scratches. Replace if damaged.</li> <li>If not damaged, sonicate the rotor, stator, packing and housing in 2- propanol for five minutes.</li> <li>Reassemble the housing in the order indicated in the figure to the right.</li> <li>Align the notch in the rotor with the mark on the shaft, and push the rotor all the way back onto the shaft.</li> <li>Insert the ring and stator ASSY onto the body. Be careful not to drop the stator.</li> <li>Insert the three hex socket head bolts and tighten them gradually with the Allen wrench.</li> <li>Reconnect the flow lines.</li> <li>Close the doors.</li> </ol> <span class="btn-link">[[LPV Rotor and Stator Replacement - SIL-30AC MP->LPV Rotor and Stator Replacement - SIL-30AC MP]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">LPV Rotor and Stator Replacement - SIL-30AC MP</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/B9TiNUVqZEo?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left; margin-bottom:25px;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: When handling the rotor, touch only the outer circumference. When returning the rotor to the instrument, wipe all sliding contact surfaces with 2-propanol. Remove the ring and housing slowly to prevent the stator and packing from dropping out of the stator ASSY.</strong> <ol> <li>Turn OFF the power switch and lower the rinse solution container to a position lower than the low-pressure valve.</li> <li>Open the doors.</li> <li>Use a wrench to remove all of the flow lines from the housing.</li> <li>Use the M4 Allen wrench to remove the three hex socket head bolts.</li> <li>Carefully, remove the stator ASSY (housing, packing, and stator) and the ring. The pieces are not connected.</li> <li>Grip the rotor securely, and pull it out of the instrument.</li> <li>Inspect the sliding surfaces of the rotor and the stator for visible nicks or scratches. Replace if damaged.</li> <li>If not damaged, sonicate the rotor, stator, packing and housing in 2- propanol for five minutes.</li> <li>Reassemble the housing in the order indicated in the figure to the right.</li> <li>Align the notch in the rotor with the mark on the shaft, and push the rotor all the way back onto the shaft.</li> <li>Insert the ring and stator ASSY onto the body. Be careful not to drop the stator.</li> <li>Insert the three hex socket head bolts and tighten them gradually with the Allen wrench.</li> <li>Reconnect the flow lines.</li> <li>Close the doors.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">HPV Rotor and Stator Removal - SIL-30AC</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/OE2Qnvx93sQ?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <ol> <li>Press [ZHOME Key] on the initial screen. The [ZHOME] screen is displayed.</li> <li>Press [enter] to move the needle up fully and to the center of the instrument.</li> <li>Turn OFF the power switch and disconnect the power cord.</li> <li>Open the doors.</li> <li>Remove the knurled thumbscrews (5) from the cooler cover.</li> <li>Pull on the cooler cover and remove it.</li> <li>Remove all of the tubing connections from the stator.</li> <li>Remove the needle seal cap using the supplied tool.</li> <li>Remove the housing and the needle seal from the stator.</li> <li>Loosen the three hex socket head bolts 90°each in sequence using the M4 Allen wrench.</li> <li>Use the long end of the Allen wrench to remove the hex socket head bolts.</li> <li>Remove the stator from the valve body.</li> <li>Carefully remove the rotor from the valve body.</li> </ol> <span class="btn-link">[[HPV Rotor and Stator Replacement - SIL-30AC->HPV Rotor and Stator Replacement - SIL-30AC]]</span> <span class="btn-link">[[HPV Rotor and Stator Removal - SIL-30AC MP->HPV Rotor and Stator Removal - SIL-30AC MP]]</span> <span class="btn-link">[[HPV Rotor and Stator Replacement - SIL-30AC MP->HPV Rotor and Stator Replacement - SIL-30AC MP]]</span> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">HPV Rotor and Stator Replacement - SIL-30AC</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/qvDg562L3mM?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left; margin-bottom:25px;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: Be sure to use the provided jig. Otherwise the stator may not uniformly contact the rotor surface, which may result in leaks or rapid wear of the rotor. CAUTION: If the stator is not leveled, the rotor and stator surfaces may be damaged and cause leaks.</strong> NOTE: When handling the rotor, try to touch only the outer circumference. After installing, wipe the sliding surface with the stator using soft cloth or paper soaked with 2- propanol. Keep the "XR IIA" marked face of the rotor toward the stator. <ol> <li>Align the notch in the rotor with the mark on the shaft, and push the rotor all the way back onto the shaft.</li> <li>Apply one or two drops of 2-propanol from a pipette or syringe onto the sliding surface in the center of the rotor.</li> <li>Place the stator on the high-pressure valve body.</li> <li>Insert the three hex head bolts into the stator and pump housing.</li> <li>Tighten them with the longer side of an Allen wrench until the bolt heads touch the housing.</li> <li>Fully insert the high-pressure rotor replacement jig between the valve body and the stator.</li> <li>Tighten the three bolts using a long leg of an Allen wrench as handle.</li> <li>Loosen the three bolts 45°. Repeat 4 times.</li> <li>Remove the jig and store it in the front door.</li> <li>Tighten the three bolts 45°. Repeat 4 times.</li> <li>Again, loosen the three bolts 45°. Repeat 4 times.</li> <li>Again, tighten the three bolts 45°. Repeat 4 times.</li> <li>Insert a new needle seal.</li> <li>Replace the needle seal cap.</li> <li>Restore the tubing at each port of the high-pressure valve.</li> <li>Reattach the sample cooler front panel.</li> <li>Secure the panel with the thumbscrews.</li> <li>Close the doors.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">HPV Rotor and Stator Removal - SIL-30AC MP</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/GChasoSuyOU?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <ol> <li>Press [ZHOME Key] on the initial screen. The [ZHOME] screen is displayed.</li> <li>Press [enter] to move the needle up fully and to the center of the instrument.</li> <li>Turn OFF the power switch and disconnect the power cord.</li> <li>Pull on the L-shaped faceplate to remove it.</li> <li>Open the key panel.</li> <li>Remove any sample racks.</li> <li>Open the rack door.</li> <li>Open the right and left doors.</li> <li>Remove the knurled thumbscrews (6) from the cooler cover.</li> <li>Pull on the cooler cover and remove it.</li> <li>Remove all of the tubing connections from the stator.</li> <li>Remove the needle seal cap using the supplied tool.</li> <li>Remove the housing and the needle seal from the stator.</li> <li>Loosen the three hex socket head bolts 90° each in sequence using the M4 Allen wrench.</li> <li>Use the long end of the Allen wrench to remove the hex socket head bolts.</li> <li>Remove the stator from the valve body.</li> <li>Carefully remove the rotor from the valve body.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">HPV Rotor and Stator Replacement - SIL-30AC MP</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/bbneC5Ij550?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left; margin-bottom:25px;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>CAUTION: Be sure to use the provided jig. Otherwise the stator may not uniformly contact the rotor surface, which may result in leaks or rapid wear of the rotor. CAUTION: If the stator is not leveled, the rotor and stator surfaces may be damaged and cause leaks.</strong> When handling the rotor, try to touch only the outer circumference. After installing, wipe the sliding surface with the stator using soft cloth or paper soaked with 2- propanol. Keep the "XR IIA" marked face of the rotor toward the stator. <ol> <li>Align the notch in the rotor with the mark on the shaft, and push the rotor fully onto the shaft.</li> <li>Moisten the surface of the rotor with one or two drops of 2-propanol.</li> <li>Place the stator on the high-pressure valve body.</li> <li>Install the hex socket head bolts (3).</li> <li>Gently tighten the three hex head bolts in sequence until the washers just touch the stator.</li> <li>Insert the high-pressure rotor replacement jig between the valve body and the stator.</li> <li>Securely tighten the three bolts using an Allen wrench 45° at a time in the sequence of A, B, C.</li> <li>Once secure, loosen the three bolts 45° at a time in sequence C, B, A (4 times).</li> <li>Remove the high-pressure rotor replacement jig.</li> <li>Tighten the three bolts 45° at a time in the sequence of A, B, C (4 times).</li> <li>Loosen the three bolts 45° at a time in sequence C, B, A (4 times).</li> <li>Tighten the three bolts 45°at a time in the sequence of A, B, C (4 times).</li> <li>Reinstall the housing and the needle seal in the stator.</li> <li>Reconnect the needle seal cap using the supplied tool.</li> <li>Reconnect all of the tubing to the stator.</li> <li>Reinstall the sample cooler front panel.</li> <li>Secure the cooler panel with the knurled thumbscrews (5).</li> <li>Close the doors and replace the sample racks.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Sample Loop Replacement - SIL-30AC MP</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/FbUIp0-mQGg?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <ol> <li>Press [ZHOME Key] on the initial screen. The [ZHOME] screen is displayed.</li> <li>Press [enter] to move the needle up fully and to the center of the instrument.</li> <li>Turn OFF the power switch and disconnect the power cord.</li> <li>Pull on the L-shaped faceplate to remove it.</li> <li>Open the key panel.</li> <li>Remove any sample racks.</li> <li>Open the rack door.</li> <li>Open the right and left doors.</li> <li>Remove the knurled thumbscrews (6) from the cooler cover.</li> <li>Pull on the cooler cover and remove it.</li> <li>Loosen the 3 mounting screws, and remove the Z-mount cover.</li> <li>Unscrew the male nut on the needle side of the sample loop using a wrench.</li> <li>Remove the male nut at port 1 of the high-pressure valve, which secures the sample loop.</li> <li>Remove the sample loop from the 4 hooks on the right side of the autosampler.</li> <li>Remove the sample loop from the square hook at the back of the Z mount, and remove it from the instrument.</li> <li>Pass the new sample loop through the square loop at the back of the Z mount.</li> <li>Attach a male nut and a ferrule to the sample loop, and secure to the joint on the needle side using a wrench.</li> <li>Attach the sample loop to the 4 hooks on the right side of the interior of the autosampler.</li> <li>Secure the sample loop to port 1 of the high-pressure valve.</li> <li>Replace the Z mount cover, and tighten with its screws.</li> <li>Replace the sample cooler front panel and close the doors.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">High-Pressure Valve Rotor and Stator</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/hpv.jpg" alt="High-Pressure Valve Rotor and Stator"></div> <h3>The rotor and stator of the high-pressure valve are consumable and need to be replaced periodically.</h3> If there are leaks in the high-pressure valve, the analysis will be adversely affected. If the high-pressure valve rotor and stator become soiled or scratched, liquid may be leaked from the high-pressure valve. Dirt or dust on the sliding surfaces may adversely affect the seal and cause liquid to be leaked. Inspect the rotor and stator for dirt or damage and clean or replace them as necessary. The rotor should be replaced after about 100,000 injections. The rotor and stator of the high-pressure valve are consumable and need to be replaced periodically. <div class="ts-question">Have the high-pressure valve rotor and stator been changed recently?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Clogged Flow Lines (1590)]]</span> <span class="btn-no">[[No->HPV Rotor and Stator Removal - SIL-30AC]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Power Cord Replacement</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/power-cord.jpg" alt="Power Cord Replacement"></div> A power supply cable is supplied with each autosampler. If the cord becomes damaged, replace it with a new one immediately. Visit <a href="https://store.shimadzu.com" target="_blank">our webstore</a> to order parts. <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Power Requirements</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/power-requirements.jpg" alt="Power Requirements"></div> Verify that the power outlet has a sufficient capacity. If capacity is insufficient, a power outage or voltage drop can occur. Power drops will affect the performance of this instrument and any other instruments that are connected to the same power supply circuit. <div class="ts-question">Does the power supply meet the power supply requirements?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Fuse Inspection - SIL]]</span> <span class="btn-no">[[No->Power Supply Requirements]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Fuse Inspection - SIL</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/fuse.jpg" alt="Fuse Inspection - SIL"></div> <h3>If the circuit has been overloaded, the fuse may blow to protect the system electronics.</h3> The glass should be clear and clean and the wire should be continuous and extend from one end of the fuse to the other. The fuse is blown and must be replaced if the glass is discolored with black or brown smudges or the wire is clearly broken or not visible at all. <ol> <li>Use a coin to open the fuse holders on the back of the autosampler.</li> <li>Remove the fuses from the fuse holders.</li> <li>Inspect each fuse by holding it up to the light.</li> <li>Insert the new fuses into the fuse holders.</li> <li>Reinstall the fuse holders.</li> <li>Close the fuse compartment cover.</li> </ol> Replacement Part - Fuse 3.15AT (5x20) P/N 072-02004-21 <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>WARNING: Turn the instrument OFF and disconnect the power supply before proceeding.</strong> <div class="ts-question">Have the fuses blown?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Fuse Replacement]]</span> <span class="btn-no"><a href="https://www.ssi.shimadzu.com/forms/tech-support/index.html?product_category=VirtualAdvisor&product_name=Nexera&page=webform_start">Contact Shimadzu</a></span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Power Supply Requirements</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/power-requirements.jpg" alt="Power Supply Requirements"></div> Ensure that the power supply meets the power supply requirements. If the power supply does not meet the specifications, the instrument may not function properly or become damaged. If the power supply does not meet specifications, take corrective action before connecting this unit to the power circuit. <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Fuse Replacement</h1> <div class="center"><iframe width="900" height="506" src="https://www.youtube.com/embed/We2QovQK9K0?rel=0" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share" allowfullscreen></iframe></div> <img class="warning" style="float: left;" src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/virtual-advisor-warning-icon.png" alt="warning-icon"><strong>WARNING: Move the needle to the [Z HOME] position, turn the instrument OFF and disconnect the power supply before proceeding.</strong> <ol> <li>Use a coin to unlock the fuse holders.</li> <li>Remove the fuse holders.</li> <li>Remove the old fuses from the fuse holders.</li> <li>Insert the new fuses into the fuse holders.</li> <li>Reinstall the fuse holders.</li> <li>Use a coin to lock the fuse holders into place.</li> </ol> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Solvent Choice</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/solvent-choice.jpg" alt="Solvent Choice"></div> Choosing the correct solvents of suitable quality is the first step in the process. It is also important to use a consistent solvent supplier, because solvents from different suppliers may vary in purity specifications. Always use solvents and reagents (including water) that are specified HPLC grade or better. <span class="btn-link">[[Solvent Preparation->Solvent Preparation]]</span> <span class="btn-link">[[Solvent Delivery->Solvent Delivery]]</span> <span class="btn-link">[[Solvent Mixing->Solvent Mixing]]</span> <div class="ts-question">Is the mobile phase composition changing?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Mobile Phase Troubleshooting (1664)]]</span> <span class="btn-no">[[No->Needle Seal 2 (1665)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Mobile Phase Troubleshooting</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/mixer.jpg" alt="Mobile Phase Troubleshooting"></div> There are a number of ways to solve mobile phase composition problems. If the problem is a mixing issue, install a mixing chamber or premix the mobile phase. If the problem is an air bubble in the solvent delivery system, degas all of the solvents that are included in the mobile phase. Solvent purity may also affect the analyte retention time. Ensure that the solvent used for mobile phase is equivalent to the historical analyses. If the solvent purity is the problem, the improvement options include changing the solvent type or increasing the purity level of the solvents <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Solvent Preparation</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/solvent-prep.jpg" alt="Solvent Preparation"></div> Whether preparing premixed isocratic mobile phases or non-mixed gradient solvents, care should be taken in the preparation process. Use caution to ensure that clean containers are used for solvent reservoirs. Employ a methodical process for buffer preparation to ensure repeatable preparations. All solvents should be filtered through a 0.45 µm (or smaller) pore size. Incorporate a thorough degassing process in the preparation process. <span class="btn-link">[[Solvent Choice->Solvent Choice (1601)]]</span> <span class="btn-link">[[Solvent Delivery->Solvent Delivery]]</span> <span class="btn-link">[[Solvent Mixing->Solvent Mixing]]</span> <div class="ts-question">Is the mobile phase composition changing?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Mobile Phase Troubleshooting (1664)]]</span> <span class="btn-no">[[No->Clogged Flow Lines (1606)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Solvent Delivery</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/lc-press-display.jpg" alt="Solvent Delivery"></div> It is important to use the designated solvent delivery type. Results may vary if results are compared from systems that incorporate different pump types. Differences in high-pressure and low-pressure gradient elution processes make it impossible to interchange them. Be sure to use the correct pump type for the application as specified in the method. <span class="btn-link">[[Solvent Choice->Solvent Choice (1601)]]</span> <span class="btn-link">[[Solvent Preparation->Solvent Preparation]]</span> <span class="btn-link">[[Solvent Mixing->Solvent Mixing]]</span> <div class="ts-question">Is the mobile phase composition changing?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Mobile Phase Troubleshooting (1664)]]</span> <span class="btn-no">[[No->Clogged Flow Lines (1606)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Solvent Mixing</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/solvent-mix.jpg" alt="Solvent Mixing"></div> It is necessary to incorporate a solvent mixing chamber for any gradient elutions. Mixing chambers create a homogeneous mixture from multiple solvent reservoirs. These chambers are necessary in any system used to blend solvents, even those performing isocratic separations. <span class="btn-link">[[Solvent Choice->Solvent Choice (1601)]]</span> <span class="btn-link">[[Solvent Preparation->Solvent Preparation]]</span> <span class="btn-link">[[Solvent Delivery->Solvent Delivery]]</span> <div class="ts-question">Is the mobile phase composition changing?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Mobile Phase Troubleshooting (1664)]]</span> <span class="btn-no">[[No->Clogged Flow Lines (1606)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Rinse Solution</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/bottle.jpg" alt="Rinse Solution"></div> Although the injection process may still function if the rinse solution runs dry, the reproducibility will suffer greatly. When the rinse solution is used up, air may be drawn into the injection tubing. Air in the injection tubing interferes with the reproducibility of the injection process. In such cases, the autosampler must be manually purged. Examine the rinse solution reservoir. <div class="ts-question">Is the volume of rinse solution adequate for the analysis?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Rinse Volume 2 (1659)]]</span> <span class="btn-no">[[No->MANUAL PURGE (10479)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Rinse Volume</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/rinse-port.jpg" alt="Rinse Volume"></div> <h3>Depending on the sample matrix, it may be necessary to increase the volume of rinse solution used to rinse the injection needle and flow path.</h3> Depending on the viscosity of the sample solution, sample carryover from one injection to another can be a problem in HPLC analysis. The most common way of reducing this affect is by rinsing the sampling needle between samples. The Shimadzu Prominence series HPLC autosampler has a number of improvements that greatly minimize this affect, such as the addition of a specially coated needle and a change in the injection port design. It is also possible to add another rinse pump to the SIL-20 autosamplers to add the capability of an active rinse of the outer surface of the needle. Shimadzu autosamplers provide for a dip rinse of the outer surface of the needle. Now, with the addition of the Additional Rinse Pump it is possible to use two different rinse solvents and perform both the dip rinse and the active rinse. <div class="ts-question">Is the RINSE VOLUME parameter adequate for the sample type?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Solvent Choice (1662)]]</span> <span class="btn-no">[[No->RINSE MODE Parameter]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Needle Seal</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/needle-seal.jpg" alt="Needle Seal"></div> <h3>The needle seal is a consumable part and needs to be replaced periodically.</h3> The needle seal provides a leak-free injection port for introducing the sample into the HPLC system. The needle seal is a consumable item for the Prominence series of autosamplers. The needle seal should be replaced after about 40,000 injections. Use the NDL SEAL USED function (Maintenance Information group) to monitor the needle seal usage. Use the NDL Seal alert function (Calibration Support group) to set the level of needle seal usage that will cause an alarm to sound. The needle seal is a consumable part and needs to be replaced periodically. <div class="ts-question">Has the needle seal been replaced recently?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Flow Line Leaks 2 (1667)]]</span> <span class="btn-no">[[No->Needle Seal Replacement - SIL-30AC]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Flow Line Leaks</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-pump/flow-lines.jpg" alt="Flow Line Leaks"></div> A leak in the flow line before the detector flow cell could explain the absence of sample peaks. Visually inspect the flow line from needle to flow cell looking for signs of a leak: <ul> <li>Puddles</li> <li>Drips</li> <li>Crystallization of buffer</li> </ul> <div class="ts-question">Are the flow line connections leaking?</div><div class="buttons"> <span class="btn-yes">[[Yes->Tubing - Flow Line Connections (1954)]]</span> <span class="btn-no">[[No->Room Temperature (1669)]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Room Temperature</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/column-temp.jpg" alt="Room Temperature"></div> <ul> <li>Upward drifting baselines usually result from an inadequately equilibrated column. Allow the column to equilibrate until the baseline stabilizes.</li> <li>Downward drifting baselines are often caused by not allowing the lamp to heat sufficiently. Allow the lamp to heat for 30 minutes before examining the baseline.</li> <li>A meandering baseline may be related to the cyclical temperature fluctuation of the column heating device.Use of a column oven can reduce the magnitude of this temperature cycling.</li> <li>A wandering baseline is often related to ambient temperature fluctuations or contaminants eluting from a column. Thermally isolate the column and other system components that can be affected by temperature changes, and wash the column thoroughly.</li> </ul> <div class="ts-question">Is the room temperature fluctuating?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Column Insulation (1689)]]</span> <span class="btn-no"><a href="https://www.ssi.shimadzu.com/forms/tech-support/index.html?product_category=VirtualAdvisor&product_name=Nexera&page=webform_start">No</a></span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Room Temperature</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/column-temp.jpg" alt="Room Temperature"></div> <ul> <li>Upward drifting baselines usually result from an inadequately equilibrated column. Allow the column to equilibrate until the baseline stabilizes.</li> <li>Downward drifting baselines are often caused by not allowing the lamp to heat sufficiently. Allow the lamp to heat for 30 minutes before examining the baseline.</li> <li>A meandering baseline may be related to the cyclical temperature fluctuation of the column heating device.Use of a column oven can reduce the magnitude of this temperature cycling.</li> <li>A wandering baseline is often related to ambient temperature fluctuations or contaminants eluting from a column. Thermally isolate the column and other system components that can be affected by temperature changes, and wash the column thoroughly.</li> </ul> <div class="ts-question">Is the room temperature fluctuating?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Column Insulation (1670)]]</span> <span class="btn-no">[[No->Column Performance (1671)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Column Performance</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/column-param.jpg" alt="Column Performance"></div> It is necessary to use a column logbook to record the performance of the column. Record the initial column parameters such as pressure, analyte retention time, Resolution (R), Plate count (N) and Tailing Factor (T). By periodically recording this information, it is possible to determine when a column needs to be flushed, regenerated or retired. <div class="ts-question">Has the column performance deteriorated?</div> <div class="buttons"><span class="btn-yes">[[Yes->Column Regeneration (1311)]]</span> <span class="btn-no">[[No->Solvent Choice (1601)]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Solvent Choice</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/solvent-choice.jpg" alt="Solvent Choice"></div> Choosing the correct solvents of suitable quality is the first step in the process. It is also important to use a consistent solvent supplier, because solvents from different suppliers may vary in purity specifications. Always use solvents and reagents (including water) that are specified HPLC grade or better. <span class="btn-link">[[Solvent Preparation->Solvent Preparation]]</span> <span class="btn-link">[[Solvent Delivery->Solvent Delivery]]</span> <span class="btn-link">[[Solvent Mixing->Solvent Mixing]]</span> <div class="ts-question">Is the mobile phase composition changing?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Mobile Phase Troubleshooting (1664)]]</span> <span class="btn-no">[[No->Clogged Flow Lines (1606)]]</span><span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Clogged Flow Lines</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/flow-sil.jpg" alt="Clogged Flow Lines"></div> The autosampler flow line is detailed above. Ensure that the flow lines are plumbed correctly. Please examine these flow lines and connections for blockage or leaks. <div class="ts-question">Are the flow lines in the injection flow path clogged or partially blocked?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Tubing Replacement (1545 / 1591)]]</span> <span class="btn-no"><a href="https://www.ssi.shimadzu.com/forms/tech-support/index.html?product_category=VirtualAdvisor&product_name=Nexera&page=webform_start">Contact Shimadzu</a></span> <span class="btn-back"><<back "Back">></span> </div> </div>

<div class="container"><h1 class="title">Column Performance</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-the-autosampler/column-param.jpg" alt="Column Performance"></div> It is necessary to use a column logbook to record the performance of the column. Record the initial column parameters such as pressure, analyte retention time, Resolution (R), Plate count (N) and Tailing Factor (T). By periodically recording this information, it is possible to determine when a column needs to be flushed, regenerated or retired. <div class="ts-question">Has the column performance deteriorated?</div> <div class="buttons"><span class="btn-yes">[[Yes->Column Regeneration (1958)]]</span> <span class="btn-no">[[No->Measuring Pump (1646 / 1571 /1673)]]</span> <span class="btn-back"><<back "Back">></span> </div> </div>