Experience new benchmarks the nexera series is a family of hplc systems that. Use table 1 page 5 to determine which components may be causing the trouble. This information offers and provides you with technical and support information relating to hplc, lcms and gcms like a basic knowledge. Pdf the developments in column packing technology and suitable equipment. Problems can arise in each component and can affect the overall system performance. Software documentation additional software documentation includes the following. Troubleshooting hplc instrumentation and separations require a.
Quick reference tables if you have a problem you need to solve now, use these troubleshooting tables. Two other modes of chromatography in which the stationary phase is a solid are classified differently from lsc and gsc because of the unique nature of their separation processes. To introduce and explain the concept of chromatographic resolution rs. Lc resources is your resource for information on all aspects of hplc and uhplc. High pressure liquid chromatography 20 chem 4 introduction chromatography can be described as a mass transfer process involving adsorption using a nonpolar stationary phase and a mobile polar phase titrating through the column.
An alternative for people with smart phones is the thermo scientific hplc troubleshooting guide an app for iphone, android, and windows platforms. If high back pressure still appears, then the blockage is in the system. If the pressure is normal, there is a problem with the column. Hplc troubleshooting for column back pressure,column leaks, changing baseline, tailing peaks, and pump and detector problems. Phenomenex offers seminars, as well as a complete line of reference books on hplc. Scientific hplc troubleshooting guide poster,4 where all troubleshooting suggestions are available at a glance and a useful suggestion can be found much faster. The article is intended to aid in troubleshooting problems that occur more often with the high performance liquid chromatography hplc system during routine analysis. 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.
You should perform regular maintenance rather than waiting for a problem to occur. High performance liquid chromatography hplc is one of the most widespread analytical and preparative scale separation techniques used for both scientific investigations and industrial and biomedical analysis. The important segments of hplc system at the same, whether we use a modular system or a more sophisticated unit. Shimadzu hplc systems demonstrate high reliability, with outstanding performance, such as ultralow carryover and exceptional area reproducibility, and superior data quality. Troubleshooting hplc instrumentation and separations require a fundamental understanding of how the instrument functions and how the separation works. Snyder, troubleshooting lc systems, humana press, nj 1989. Read online hplc troubleshooting guide sigmaaldrich book pdf free download link book now. We would like to show you a description here but the site wont allow us. A blank page in the web browser a red x icon a brokenlink indicator, such as a red square, a blue triangle, or a blue circle error. Quantitative methods july 11, 2019 fundamental guide to gc 4.
So that it can be easily known from file\folder name which run it carry. Sep 21, 2005 i would like to know that what procedure you follow for naming of files of hplc run. Lc tips and tricks seminar series page 2 trouble shooting steps you have recognized there is a problem. Comprehensive guide to successfully troubleshooting hplc columns. Hplc troubleshooting hints and tips chromatography. When a problem occurs, it is advisable to perform a quick visual check of the instrument and column. Troubleshooting in your installation and maintenance guide.
Carryover carryover is the appearance of a peak in a blank injection after a high concentration of sample. High performance liquid chromatography hplc is a chromatographic technique used to separate a mixture of compounds in analytical chemistry and biochemistry with the purpose of identifying, quantifying or purifying the individual components of the mixture. Kirkland, introduction to modern liquid chromatography. Hplc lthough hplc method has been improved by advances in column technology and instrumentation, problems still arise. Hplc troubleshooting and method optimization everybody who ever worked in analytics knows that the work doesnt always run smoothly. There are a number of reference sources that can give you guidance in problem solving. There are five major of symptoms to help you quicklyidentify the source of the problems. Snyder, troubleshooting lc systems, humana press, nj. When you press pump onoff your pump initializes the mechanical parts to ensure they are functioning correctly. Jun 04, 2018 every hplc system consists of the same important components, no matter if its a modular system or a specialized allinone unit. A representation of the tray is shown on the right. Place your samples in a row in the tray keep them together as a group. In addition, they are carefully designed for broad applicability and easy operation.
If you use a manual injector, make sure that your syringe is properly cleaned. Injector and injection solvents the injector rapidly introduces the sample into the system with minimal disruption of the solvent flow. Hplc column troubleshooting what every hplc user should. The adobe acrobatreader that is running cannot be used to view pdf files in a web browser. Hplc troubleshooting guide thermo fisher scientific.
Primary aim is to focus the commonly encountered problems along with more indepth information to help the reader understand. A process of elimination will usually enable you to. Tips and tricks of hplc system troubleshooting agilent technologies, inc. Hplc troubleshooting and maintenance a logical approach to hplc troubleshooting and maintenance is fully explored within this oneday course. Pdf highperformance liquid chromatography is one of the most used analytical techniques in industry. This chapter gives an overview of hplc system maintenance and summarizes strategies and guidelines for hplc troubleshooting. Tip electronic versions of these manuals are available as pdf portable document format files. If a cartridge system is being employed then an integral cartridge holder must also be. Frequently encountered troubleshooting problems are classified into four categories pressure, baseline, peak, and data. Highperformance liquid chromatography hplc scope instrumentation eluants, injectors, columns modes of hplc partition chromatography adsorption chromatography ion chromatography size exclusion chromatography. It is provided in the form of a webhelp file that can be read with any browser.
Identify and correct common hplc problems to restore column performance. In an hplc system, problems can arise from many sources. To isolate the source of a baseline irregularity, it is important to determine whether the problem lies with the fluid path, detector or electrical connections. Hplc troubleshooting guide sigmaaldrich pdf book manual. Lightweight application to help beginner hplc users with hplc troubleshooting. If same, consider starting a zfresh tune page since tune files can become corrupt.
If you have the original source file that the pdf was created from, recreate the pdf. In comparison to former days, technology and instrumentation have been improved but typical problems still occur. Troubleshooting guide hplc before starting any troubleshooting, whether it is related to instruments or columns, it is essential that safe laboratory practices be observed. How to make an hplc sequence file after you have placed your samples in vials, a ta will assist you to place them in an autosampler tray. Hplc troubleshooting guide phenomenex uhplc, hplc, spe and gc. Problems often appear when you need them the least. Troubleshooting typial causes of failure sensitivity ion transmission is drastically effected by source tuning parameters. Get help with peak problems peak splitting, peak tailing, peak fronting, etc. This is a compilation of john dolans lc troubleshooting columns from lcgc magazine from 1983 through september 2016. This will pick up leaks, loose or disconnected tubing, changes. Slide 5 hplc components pump injectorautosampler column detector data systemintegrator all of these components can have problems and require troubleshooting. Chromeleon help and documentsthe chromeleon help provides extensive. Even excellently maintained devices can suddenly show changes.
Hplc system components problems can be related to all components in the system page 4 categories of column and system problems a. Hplc column troubleshooting what every hplc user should know. Useful data is provided throughout the book, such as. Hplc troubleshooting guide phenomenex uhplc, hplc, spe. All books are in clear copy here, and all files are secure so dont worry about it. How to diagnose and solve chromatographic problems poster pdf available february 2018 with 5,436 reads how we measure reads. A bench manual, appuyez sur extrait gratuit ou acheter.
All other ham lc columns all other sho lc columns ax pak chi chiradex chiral cdph curosil taxol hypersil lichrosorb nucleodex nucleosil ab nucleosil hd nucleosil plgel sumichiral pdf link. At the end of each chapter there is a list of references andor further reading which will help the reader to develop their expertise in the technique. Changing the mass of injected sample is a simple way to help elucidate the characteristics of a chromatogram. Hplc troubleshooting ebook page 7 hplc column maintenance prevention is better than cure guard columnsdisks a guard column is a short column typically 1020 mm in length that is packed with the same stationary phase material as the analytical column. Hplc troubleshooting guide category troubleshooting matrix method hplc, uhplc keywords troubleshooting, hplc and uhplc problems, column care and use analytes id vsp0003n, 0911 summary in hplc or uhplc numerous problem can arise. In addition to providing an introduction to hplc for pharmaceutical analysis it is intended that this book will be a useful resource. Use table 1 to determine which components may be causing the trouble.
Troubleshoot viewing pdf files on the web adobe inc. 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. A guide to problemsolving in high performance liquid. Locate the table for the type of problem you have, find the possible cause and use the short description of the solution or the crossreference to the main body of the guide to help you identify and solve your problem quickly. To open and read the pdf files, adobe reader or adobe acrobat is required. Notice the markings 112 on the top and ah on the left side. Guide to lc troubleshooting guide to lc troubleshooting is a special project supplement produced by lcgc europe chester, uk and edited by lcgc column watch editor ronald majors agilent technologies inc. In reader or acrobat, choose file save as and give the pdf file a new name. The reproduction, modification, storage in a retrieval system, or retransmission, in any. It describes common maintenance procedures that can be performed by the user.
Like file naming, date of run, compound name, batch name. Hplc troubleshooting and maintenance crawford scientific. When you turn on line power, your pump checks the controller functions. Troubleshooting hplc instrumentation and separations. Emitter position and spray stability is the both the easiest way to have a problem and to fix one. This can be achieved by following the simple steps. Troubleshooting troubleshooting is a form of problem solving, often.
A process of elimination will usually enable you to pinpoint the specific cause and correct the problem. In figure 1a, a shoulder on the front of the main peak became a second peak figure 1b when we reduced the amount of sample. Resolution in hplc pdf in order to justify its classifichtion as a high resolution technique, hplc should show an improvement in efficiency by a factor of at least 10, should be easily. With this troubleshooting guide, we provide help for solving typical and frequently found problems in hplc and uhplc. The active component of the column, the sorbent or the stationary phase, is typically a granular. As mentioned above, sideport needles are generally free of septumrelated problems, but this needle design is not in widespread use. Download hplc troubleshooting guide sigmaaldrich book pdf free download link or read online here in pdf. How to identify, isolate, and correct the most common hplc problems.
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