Nexera HPLC Troubleshooting

Browser lacks capabilities required to play.

Upgrade or switch to another browser.

Loading…

Config.debug = false;

<div class="container"><h1 class="title">Troubleshooting Split Peaks</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-split-peaks/Split.jpg" alt="Troubleshooting Split Peaks"></div> The most common reasons for peaks to split are strong sample solvent, blocked column frits or column voids. When all of the peaks are splitting, it is an indication that there is a void in the column. If only the early eluting peaks are deformed or splitting, it is usually an indication that the sample solvent is too strong for the mobile phase. This occurs when the sample is dissolved in a stronger solvent (i.e. 100% MeOH) than the mobile phase (i.e. 50:50 MeOH:Water). <div class="ts-question">Are all of the peaks in the chromatogram split?</div> <div class="buttons"> <span class="btn-yes">[[Yes->Cutting Resin]]</span> <span class="btn-no">[[No->Strong Sample Solvent]]</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-split-peaks/Tubing_FlowLineConnections.jpg" alt="Tubing - Flow Line Connections"></div> List of Steps: <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><br> <span class="btn-link">[[Cutting Resin->Cutting Resin]]</span><br> <span class="btn-link">[[Cutting SS->Cutting SS]]</span><br> <span class="btn-link">[[Tighten SS->Tighten SS]]</span><br> <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-split-peaks/TubingKnife.jpg" alt="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">[[Tubing-Flow Line Connections->Tubing-Flow Line Connections]]</span><br> <span class="btn-link">[[Cutting SS->Cutting SS]]</span><br> <span class="btn-link">[[Tighten SS->Tighten SS]]</span><br> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div>

<div class="container"><h1 class="title">Tubing - Cutting Stainless Steel</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-split-peaks/TubingSS.jpg" alt="Cutting Stainless Steel"></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><br> <span class="btn-link">[[Tubing-Flow Line Connections->Tubing-Flow Line Connections]]</span><br> <span class="btn-link">[[Tighten SS->Tighten SS]]</span><br> <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div>

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

<div class="container"><h1 class="title">Strong Sample Solvent</h1> <div class="center"><img src="https://www.ssi.shimadzu.com/sites/ssi.shimadzu.com/files/support/virtual-advisors/nexera/troubleshooting-split-peaks/StrongSampleSolvent.jpg" alt="Strong Sample Solvent"></div> The sample solvent is probably too strong for the mobile phase. Often when the sample is dissolved in a stronger solvent (i.e. 100% MeOH) than the mobile phase (i.e. 50:50 MeOH:Water), the peak shape will be affected. In this situation, fronting shoulders are more noticeable in the earlier eluting peaks than in the later peaks. Use mobile phase for the sample solvent or choose a sample solvent that more closely matches the properties of the mobile phase. Decrease the injection volume by 1/2 and make 3-5 repeat injections. If decreasing the injection volume improves the peak shape, it is an indication that the sample solvent is stronger than the mobile phase. If decreasing the injection volume does not improve the peak shape, it is an indication of a probable mixing problem. Add a mobile phase mixing chamber or increase the mixing volume. <div class="buttons"> <span class="btn-back"><<back "Back">></span> </div>