Software designed to track inventories, manage schedules, aggregate data, provide resource visibility, and integrate with other lab systems. Troubleshooting troubleshooting is a form of problem solving, often. Veronika meyer, pitfalls and errors of hplc in pictures, third edition, 20. For example, replacing pump seals at regular intervals should eliminate. Hplc troubleshooting hints and tips chromatography. Highperformance liquid chromatography hplc scope instrumentation eluants, injectors, columns modes of hplc partition chromatography adsorption chromatography ion chromatography size exclusion chromatography. Many problems in an lc system show up as changes in the chromatogram. 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. Use table 1 to determine which components may be causing the trouble. The important segments of an hplc system are the same, whether you use a modular system or a more. Hplc system components problems can be related to all components in the system page 4 categories of column and system problems a. In this guide we offer you a systematic means of isolating, identifying, and correcting many typical problems.
Hplc troubleshooting monitor at least one peak in one injection plate count peak width relative to rt, peak asymmetry, retention time andor retention parameter relative retention time for critical pair of analytes. Hplc lthough hplc method has been improved by advances in column technology and instrumentation, problems still arise. Quantitative methods july 11, 2019 fundamental guide to gc 4. Injector and injection solvents the injector rapidly introduces the sample into the system with minimal disruption of the solvent flow. If a cartridge system is being employed then an integral cartridge holder must also be. Hplc errors for the real optimum under changed mobile phase flow speeds of figure 1 on this page below. Reproduce the production test chromatogram using conditions noted on the quality assurance report that was shipped with the column to assess column performance. Some of these can be solved by changes in the equipment. Sadek troubleshooting hplc systems a bench manual 2000 isbn 0471178349 u. Labsolutionshplc instrument communication settings and. Symptom solution decreasing retention times column usage can affect capacity. No pressure 20 waters corporation 10 air in system o prime the pump methanol or ipa to remove air air in solvent lines. Lc tips and tricks seminar series page 2 trouble shooting steps you have recognized there is a problem. Mar 11, 2019 i am using an agilent 1260 infinity ii prime lc comprised of.
The last twentyfive years have seen a dramatic rise in the use of high performance liquid chromatography hplc in laboratories worldwide. Page 1 page 2 page 3 page 4 page 5 table of contents page 6 page 7 page 8 page 9 page 10 page 11 page 12 page installation page 14 page 15 page 16 power supply and wiring page 17 page 18 page 19 gas supply plumbing page 20 page 21 page 22 page 23 page 24 page 25 page 26 page 27 page 28 page 29 before use page 30 page 31 page 32. In this course, although we do consider problems arising from instrumentation and the need for preventative maintenance, the main emphasis is on solving problems in the chromatography. Use hplc grade solvents, high purity salts, and additives. The tz value is a way out for correct flow and temperature optimization in hplc. Variation in retention is generally the result of eluent preparation errors or pump issues. How to diagnose and solve chromatographic problems. Neue hplc columns theory, technology, and practice 1997 isbn 0471190373 l.
To isolate and repair specific problems related to your apparatus, use the troubleshooting and maintenance sections of the operation manual. Useful data is provided throughout the book, such as. Lc chromatography troubleshooting hplc columns in india. Troubleshooting guide hplc before starting any troubleshooting, whether it is related to instruments or columns, it is essential that safe laboratory practices be observed. Gc troubleshooting gc troubleshooting hints and tips. An alternative for people with smart phones is the thermo scientific hplc troubleshooting guide an app for iphone, android, and windows platforms. 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. Diagnose and resolve problems associated with hplc. Many gc problems can be prevented if the column is properly installed and gc is maintained routinely.
The advantage with calculating standard deviation is that for random errors, they follow the statistical normal distribution model so we can show that 68. Hp laserjet pro p1606dn printer troubleshooting hp. 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. Selecting the proper column type and mobile phase are keys to good chromatography. Column troubleshooting peak shape problems column destroyed secondary interactions incorrect sample solvent. Pressure problems fall into one of three categories. Waters prep series installation and maintenance manual pdf. Troubleshooting hplc systems provides analysts as well as laboratory technicians and managers with a readily accessible and immensely useful guide to the new generation of hplc equipment and techniques. Many lc problems can be prevented with routine preventive mainte nance. Page 112 autoinjector troubleshooting continued symptom possible cause corrective action reproducibility errors syringe contains an air remove bubble. The important segments of hplc system at the same, whether we use a modular system or a more sophisticated unit. Understand how hplc works and the role of each component in a hplc system. System pressure problems low pressure check if something has changed column, mobile phase, temperature, method if nothing has changed, check for leaks. For example, replacing septa or liner at regular intervals and keeping the injector and detector clean and wellmaintained shoud solve many problems.
Pdf highperformance liquid chromatography is one of the most used. Practical hplc method development this course is very different from the hplc equipment servicing course. I am using an agilent 1260 infinity ii prime lc comprised of. Many liquid chromatography lc problems can be prevented with routine maintenance.
From my little experience i learnt that it may be due to air trapped in. If youre having problems signing in, first make sure youre using the latest version of skype, your system meets the minimum requirements for running skype. 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. Hplc troubleshooting and maintenance crawford scientific.
This guide is intended as an aid in troubleshooting problems that occur in the day to day running of your. 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. You should perform regular maintenance rather than waiting for a problem to occur. 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. Understand how problems can arise in the individual components of a hplc system. Experience new benchmarks the nexera series is a family of hplc systems that.
Quick reference tables if you have a problem you need to solve now, use these troubleshooting tables. Hplc injection is a technique used to load the sample into the hplc system. 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. Troubleshooting and diagnostics troubleshooting system pressure problems. Hplc troubleshooting and maintenance a logical approach to hplc troubleshooting and maintenance is fully explored within this oneday course. In an hplc system, problems can arise from many sources. The third edition of this popular problemsolving guide for this widelyused method includes eleven completely new examples and several updated ones, adding up to over 100 potential pitfalls and errors in hplc. How do i inhibit bubbles from getting into the flow cell. High pressure cracks cell win dow, to detector manual to replace. A process of elimination will usually enable you to. Tips and tricks of hplc system troubleshooting agilent technologies, inc. Stuart jones hplc academy hplc troubleshooting course.
There are five major of symptoms to help you quicklyidentify the source of the problems. The most popular injector is the rheodyne injector. Hplc troubleshooting guide phenomenex uhplc, hplc, spe and gc. There are many areas in a hplc instrument that give rise to system and.
Each example is presented on a double page with the text on the lefthand and a fi gure on the righthand side, true to the motto a picture says more than a thousand words. Pitfalls and errors of hplc in pictures, third edition dr. Although hplc method development has been improved by advances in column technology and instrumentation, problems still arise. Hplc troubleshooting pdf proteins and proteomics opening. Not enough solvent in bottle o replace the solvent bottle problem with check valves. Flush column with strong mobile phase at end of each run see 3. Degas mobile phase before use, sparge with helium during use.
If high back pressure still appears, then the blockage is in the system. This information offers and provides you with technical and support information relating to hplc, lcms and gcms like a basic knowledge. In addition, they are carefully designed for broad applicability and easy operation. Service contracts, on demand repair, preventive maintenance, and service center repair. Troubleshooting and maintenance services are provided by the supplier as and when required. We have been experiencing some pressure problems on our hplc lately and the problem is mostly fluctuations in the pump head.
If necessary, clean cell with 1n hno 3 never use hcl. Labsolutions workstation introduction and new hplc instrument communication settings and hardware configuration guide. Flush injector between analyses late eluting peak from previous injection 1. The latter is still underdeveloped in hplc but often very helpful. The detailed training of hardware and software provided to users. In addition to providing an introduction to hplc for pharmaceutical analysis it is intended that this book will be a useful resource. Scientific hplc troubleshooting guide poster,4 where all troubleshooting suggestions are available at a glance and a useful suggestion can be found much faster. A process of elimination will usually enable you to pinpoint the specific cause and correct the problem. Hplc troubleshooting guide thermo fisher scientific.
Hplc troubleshooting for column back pressure,column leaks, changing baseline, tailing peaks, and pump and detector problems. Use table 1 page 5 to determine which components may be causing the trouble. If the pressure is normal, there is a problem with the column. Implement measures which prevent problems occurring. Aug 01, 2016 labsolutions workstation introduction and new hplc instrument communication settings and hardware configuration guide. Comprehensive guide to successfully troubleshooting hplc columns. Lc chromatography troubleshooting how do i remove bubbles from the flow cell. Glajch practical hplc method development 1997 isbn 047100703x. Pitfalls and errors of hplc in pictures, third edition. Use a systematic problemsolving approach to hplc troubleshooting.
Qualification of hplc systems for hardware and software at the site is carried out in the presence of system custodian. This process is done using a specially designed injection system which is a separate and prominent part of the hplc system. Distorted peaks will cause integration or resolution problem indication that optimal column performance is not being attained. The sample and liquids used in hplc and lcms are chemicals which. Every lcms user can work to hone their skills in troubleshooting and maintenance. Troubleshooting sources, vacuum, and system communications best practices setup, power, gases, logbooks and pm increase uptime. Quat pump g7104c, samplerg7129c,column oven g7a and dad g7115a. The best way to avoid getting bubbles in the flow path is to pump the mobile phase through a degasser module.
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