Introduction hplc lthough hplc method has been improved by advances in column technology and instrumentation, problems still arise. Waters 2690 quick start manual pdf download manualslib. This guide sets forth procedures for unpacking, installing, using, maintaining, and troubleshooting the waters alliance e2695 separations module. Some troubleshooting work has already been performed. A primer sue abbatiello, the broad institute tom blau, ion technology support, inc. Troubleshooting the 26905 user maintenance of the 26905 spare parts. Problems affecting overall system performance can arise in each component. It also describes detector optics and the principles of spectral contrast used in the millennium 32 software for. Primary aim is to focus the commonly encountered problems along with more indepth information to help the reader understand. Not enough solvent in bottle o replace the solvent bottle problem with check valves.
Waters hplc troubleshooting guide pdf but really, you should be prepared to do this application note. A process of elimination will usually enable you to pinpoint the specific cause and correct the problem. Troubleshooting guide hplc before starting any troubleshooting, whether it is related to instruments or columns, it is essential that safe laboratory practices be observed. Tips and tricks of hplc system troubleshooting agilent. To remove these contaminants, pass the deionized water through activated charcoal or a preparative c18 column. In an hplc system, problems can arise from many sources. When that was switched out we were not able detail of the problem. Waters 2690 separations module operators guide 34 maple street milford, ma 01757 wat55302tp, revision 2. A process of elimination will usually enable you to. The important segments of an hplc system are the same, whether you use a modular system or a more sophisticated unit. Use table 1 page 5 to determine which components may be causing the trouble. Nov 12, 2017 operators guide 252 hplc, or fax 508 872 1990. By doing so, this manual serves as a solid help for new empower users to get started and as a reference for those users already experienced in the routine use of the cds software. Uwe neue from the waters corporation, was developed to identify, explain, and solve some of the most common sources of laboratory problems in the areas of hplc and column technology.
Do not start to clean the uplc hplc system until you prevent future contamination new andor clean bottles never wash uplc hplc glassware in the dishwasher 20 waters corporation 36 rinse only with the hplc grade solvents if the contaminant is known, what determines your choice of solvents. The waters 2690 separations module quick start guide contains basic procedural information to help you set up the waters 2690 separations module and make a run. There are over 50 topics for discussion in this guide which includ. The consistency of the alliance hplc system gives you exceptional confidence in your analytical results, daytoday and yeartoyear. Clean all solvent inlet filters in a sonic bath using 6n nitric acid, followed by water, then finally methanol. Remove the solvent line filter from the mobile phase bottle. Hplc uplc system issues can be detected early, minimizing. A quick guide to the top 10 things that must be avoided in your hplc analysis.
An lcmsms is a liquid chromatograph lc system coupled with a quadrupole mass spectrometer ms. Although hplc method development has been improved by advances in column technology and instrumentation, problems still arise. Flush with water, and then store in 7030 acnh2o for reversed phase if column will be used within four days. Calibrating to calibrate lc systems, follow acceptable calibration methods using at least five standards to generate a standard curve. It also includes appendixes for specifications, spare parts, and solvent considerations. Waters alliance e2695 operators manual pdf download.
The baseline may rise, and spurious peaks can appear as the level of the contaminated component increases. The waters acquity ultraperformance liquid chromatograph and the micromass quattro premier triple quadrupole mass spectrometer. It is also intended as a guide to best practice in systems that are free from problems. Use table 1 to determine which components may be causing the trouble. View online or download waters hplc installation and maintenance manual. The guide covers the main columns used in lc, with particular emphasis on reversedphase high performance liquid chromatography. We have 1 waters hplc manual available for free pdf download. This guide is intended for use by individuals who need to install, operate, maintain, or troubleshoot the waters alliance e2695 separations module. Waters 2489 uvvisible detector the detector can operate as a standalone unit with a chart recorder or integrator or as an integral part of a waters chromatography system. Column troubleshooting is more usually performed by reference to the chromatogram and chromatographic peak. System pressure problems low pressure check if something has changed column, mobile phase, temperature, method if nothing has changed, check for leaks. Fluctuating pressure, high delta psi with alliance.
Use the waters 1525, 1525, and 1525ef hplc pumps and the optional overview and maintenance guide, not operators guide, accompanies the instrument 9 nov 2016 4 aug 2012 operators guide. There are over 50 topics for discussion in this guide which include. The waters acquity ultraperformance liquid chromatograph. Hplc is an abbreviation for high performance liquid chromatography it has also been referred to as high pressure lc hplc has been around for about 35 years and is the largest separations technique used the history of hplc.
The waters prep series hplc system is for research use only. Aqueous gfc and nonaqueous gpc n amino acid analysis n sfc columns n hplc specialty columns for analysis of. Most of their help is suggesting which instrument would be task oi. Intended use of the waters prep series hplc system use the waters prep series hplc system to isolate and purify a wide variety of samples. The waters 2489 uvvisible detector is a twochannel ultravioletvisible uvvis detector designed for high performance liquid chromatography hplc applications. Hplc troubleshooting guide follow the four steps of trouble shooting using this guide to identify and resolve your hplc problems. Service in not 100% sure what is reason for that bubbles in detector cell could be. Hplc system components problems can be related to all components in the system page 4 categories of column and system problems a.
If you do not have the shunt cell, then we would recommend they contact their local waters engineer. In this guide we offer you a systematic means of isolating, identifying, and correcting many typical hplc problems. This guide will be useful for analysts who are looking for a summary of the most common problems encountered in hplc analysis along with troubleshooting clues on how to diagnose and solve these. Repair waters hplc error messages troubleshooting guide. Fluctuating pressure, high delta psi with alliance 2695 sm or 1515 pump wkb21017 last updated. This guide sets forth procedures for unpa cking, installing, using, maintaining, and troubleshooting the waters alliance e2695 separations module. Twitter, linkedin, facebook, youtube and our exclusive user communities. Its primary aims are to provide a logical process when troubleshooting, to maximize system. Uwe neue from the waters corporation, was developed to identify, explain, and solve some of the most. Waters hplc troubleshooting guide pdf you should try to maintain the request again. The sample and liquids used in hplc and lcms are chemicals which. Less common and lastly there could be other electrical problems such as voltage drops or issues with drivers on the personality pcb that might cause the same.
When problems arise on an hplc or uplc system, system troubleshooting can. A guide to problemsolving in high performance liquid. Hplc troubleshooting guide thermo fisher scientific. Heap exhausted in call to new what does that means. The important segments of hplc system at the same, whether we use a modular system or a more sophisticated unit. Waters alliance e2695 operators manual pdf download manualslib. In normal phase chromatography the retention times are. Lc tips and tricks seminar series page 2 trouble shooting steps you have recognized there is a problem. Hplc troubleshooting guide men common causes for abnormal chromatograms abnormal peak shapes main cause solution frequent problem peak tailing selection of column and or eluent appropriate no yes strong sample retention void existing at top of column change column andor eluent replace column peak split, shoulders adsorption of impurities in. Hplc standard operating procedure last revised march, 3, 2016 this document is intended to be a guide for operating the garner labs hplc systems and is only intended to provide instructions for users who are already generally familiar with hplc operation. Increase noise setting level until only retention times of peaks of interest are printed with retention times. This guide is organized by five major categories of symptoms to help you quickly identify the source of the problems you are encountering. Waters hplc manuals manuals and user guides for waters hplc.
Waters e2695 injection peak area repeatability problem. Using two 516inch wrenches, disconnect the tubing that goes into the inlet and outlet check valves. When problems arise on an hplc or uplc system, system troubleshooting can be a time. Integration startstop marks occur too early or late. Check obvious explanations and change only one thing at a time. The article is intended to aid in troubleshooting problems that occur more often with the high performance liquid chromatography hplc system during routine analysis.
Dear deleon, i have this message on my system, from time to time. Troubleshooting common system problems using waters neutrals. The waters engineer can go on site with the shunt tool and do that troubleshooting. The chemical and physical properties of any solvents used should be known and the material safety data sheet msds for theses solvents should be readily available. Get accurate and reproducible results with waters flagship hplc technology, which provides consistent and reliable performance whether your alliance hplc system is newly manufactured, or youre working on one that is several years old. Telephone and fax from the usa or canada, phone 800 252 hplc, or fax 508 872 1990. Waters uplc troubleshooting and diagnostics mass spectrometry. No pressure 20 waters corporation 10 air in system o prime the pump methanol or ipa to remove air air in solvent lines. Prepare fresh mobile phase and purge the solvent lines. In addition, this guide contains detailed flow charts that provide waters recommended steps to prime, equilibrate, and purge both degasser and spargebased 2690 separations modules. This guide is intended as an aid in troubleshooting problems that occur in the day to day running of your hplc system.
For other overview and maintenance guide, not operators guide, accompanies the. Waters xbridge peptide beh c18 care and use manual pdf. Tips and tricks of hplc system troubleshooting agilent technologies, inc. Appendix e 221 this section is designed to assist in troubleshooting system problems. Waters prep series installation and maintenance manual pdf. Feb 01, 2016 overcoming some of the common problems that can occur with columns and fittings in everyday use. Repair waters 2487 detector error troubleshooting guide. These clog up when people abuse the systems andor do not. Hplc system components problems can be related to all components in the system. Should you have any suggestions or just like to give feed.
Waters, thus avoiding an information overload and keeping the manual as compact as possible. Isolating hplc problems in an hplc system, problems can arise from many sources. Hplc troubleshooting guide introduction during the development of a new hplc method, there were unexpected resultsall peaks, except for the injection peak, were negative figure 1. For other locations worldwide, phone and fax numbers appear in the waters web site. Waters contact information contacting medium information internet the waters web site includes contact information for waters locations worldwide. Problems can be related to all components in the system.
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