Standard operating procedure high performance liquid. The agilent 1100 series autosampler is designed for use with other modules of the agilent 1100 series lc system, with the hp 1050 series, or with other lc systems if adequate remote control inputs and outputs are available. In an hplc system, problems can arise from many sources. The contamination can last for several sequential runs, often decreasing in amount after each injection which is a key observation. In addition to providing an introduction to hplc for pharmaceutical analysis it is intended that this book will be a useful resource. Hplc, or high performance liquid chromatography, is an analytical tool used in laboratories to detect individual solutes within a complex mixture. Agilent technologies makes no warranty of any kind with regard to this material, including but not limited to the implied warranties of merchantability and fitness for a particular purpose. Page 1 agilent 1200 series refractive index detector user manual page 2 wa r n i n g contains the complete information about this document are furnished under a license the agilent 1200 series refractive index and may be used or copied only in accor a warning notice denotes a detector. Care, maintenance, and troubleshooting oftroubleshooting of hplc columns columns and consumables edward kim apppp glications engineer january 17, 2008. User manual, quick start manual, operators quick manual. The nexera gpc system provides highly reliable and highly extensible performance in a wide variety of applications. As an agilent customer, you have complete access to over 40 years of industry knowledge and experience.
At ph 8 91% of the sample exists as protonated diphenhydramine. It is also intended as a guide to best practice in systems that are free from problems. Cleaning lower quality solvents is time consuming, and trace levels of contaminants often remain. Lc tips and tricks seminar series page 2 trouble shooting steps you have recognized there is a problem.
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. It is available as adobe reader file dance with the terms of such license. I have attached a very rough drawing of the problem. The volume between the point of mixing of solvents usually in the mixing chamber or at the proportioning valves in the liquid chromatograph and the head of an lc column. Before hplc was available, lc analysis was carried by gravitational flow of the eluent the solvent used for lc analysis thus required several hours for the analysis to be completed. View and download agilent technologies 1200 series user manual online.
Troubleshooting troubleshooting is a form of problem solving, often. Plus youll have all the agilent part numbers youll need in one easytouse reference guide. These trace contami nants can cause problems when you use a high sensitivity ultraviolet or fluorescence detector. We are having intermittent baseline issues on an 1100 hplc. Introduction we build our hplc systems from the ground up and provide our. High performance liquid chromatography hplc using agilent. Carryover, carryover, carry over, hplc, uhplc, injector, contamination, is a term used to describe a type of sample contamination which causes sample peaks to reappear in later runs which do not actually contain the sample e. Collection of lc hplc resources lc troubleshooting. Distorted peaks will cause integration or resolution problem.
For persistent problems relating to the column or your particular analysis, please contact supelcos technical service. Welcome to the agilent 2100 bioanalyzer maintenance and troubleshooting guide. A quick look at how to use this guide on page 8 explains how easy it is to use this online manual and helps you to get. The choice of column dimensions will depend on the chromatographic application. Modern hplc systems often have selfdiagnostic capabilities that help isolate the problem area within the instrument. 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. The active component of the column, the sorbent or the stationary phase, is typically a granular. If this is not available, sparge the mobile phase with helium. Quick reference tables if you have a problem you need to solve now, use these troubleshooting tables. With a wide range of technical knowhow and a neverending supply of ideas, our goal is to help you solve your most challenging application 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.
Unpacking the agilent g4240a hplcchip ms interface. We manufacture our hplc systems from the ground up and provide our customers with a wide variety of columns, consumables, and supplies built to agilent s precise speci. At ph 10 91% of the sample exists as unprotonated diphenhydramine. Hplc lthough hplc method has been improved by advances in column technology and instrumentation, problems still arise. The detector should be operated in a horizontal position. Oct 02, 2017 agilent 1200 series hplc system is available from the lab world group. Agilent maintenance guides on this page you will find manuals for the extensive agilent instrument ranges of hplc, gc, ms and spectroscopy products. Agilent hplc uhplc systems spectralab scientific inc. The pump must deliver the mobile phase at high pressures between 50 and 0 bar to overcome the resistance of the stationary phase in the column.
Acrobat reader documents pdf located on the value solution. Tips and tricks of hplc system troubleshooting agilent technologies, inc. Autosamplers are more and more used in hplc to improve the. Agilent 2100 bioanalyzer maintenance and troubleshooting. Hplc troubleshooting hints and tips more about chromatography. Here is a best of collection of links to manuals and tools for hplc on, for other instruments refer to collection of support documents on. Can anyone help with a baseline issue on an 1100 agilent hplc. Hplc troubleshooting guide phenomenex uhplc, hplc, spe and gc. Jan 02, 2019 hplc, or high performance liquid chromatography, is an analytical tool used in laboratories to detect individual solutes within a complex mixture. There are over 50 topics for discussion in this guide which includ. The 1260 infinity ii lc is the flexible instrument choice, while the 1260 infinity ii prime lc brings more operational convenience.
Lc chromatography troubleshooting how do i remove bubbles from the flow cell. Comprehensive guide to successfully troubleshooting hplc columns. Standard operating procedure high performance liquid chromatography hplc agilent technologies 1260 infinity high performance liquid chromatography hplc is a technique in analytical chemistry used to separate, identify, and quantify each component in a liquid mixture. As well as routine maintenance procedures, you will also find exploded instrument diagrams and quick lists to ensure you choose the correct parts for your repair and maintenance operations.
The autosampler is controlled from the agilent 1100 series control module or from the agilent chemstation for lc systems. Hplc troubleshooting high performance liquid chromatography. At ph 9 the sample exists as protonated and unprotonated diphenhydramine in a ratio of 1. Hplc column dimensions will affect the efficiency sensitivity, and speed of an analysis. Article from lcgc regarding check valve sticking issues when acn is used as the organic mobile phase. If the bench should carry a agilent 1100 series system, make sure that the bench is designed to bear the weight of all modules. Agilent 1200 series hplc system is available from the lab world group. This online manual provides novice and advanced users with information needed to successfully run chips with the agilent 2100 bioanalyzer. View online or download agilent technologies 1260 infinity user manual. Pressure problems fall into one of three categories. You should perform regular maintenance rather than waiting for a problem to occur. Agilent technologies 1260 infinity pdf user manuals. How to diagnose and solve chromatographic problems.
Hplc troubleshooting guide phenomenex uhplc, hplc, spe. Youll discover important maintenance procedures and troubleshooting tips. Download file pdf hplc troubleshooting guide agilent file type hplc troubleshooting guide agilent file type as recognized, adventure as well as experience practically lesson, amusement, as with ease as conformity can be gotten by just checking out a ebook hplc troubleshooting guide agilent file type as a consequence it is not directly done, you. Hplc stands for high performance liquid chromatography. This guide is intended as an aid in troubleshooting problems that occur in the day to day running of your hplc system. 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. Impurities in the acn accumulate in the seat of most check valves and cause the valve to stick or not seal correctly. Agilent 11001200 hplc chemstation operation course number h4033a laboratory manual. Were also a great resource for all types of applications in fact, were developing new ones every day. Jun 30, 2017 here is a best of collection of links to manuals and tools for hplc on agilent. For performance, selectivity and guaranteed reproducibility. Agilent maintaining your agilent 1100 series hplc system.
Final tips and tricks hplc troubleshooting 2 agilent. Agilent 1200 series standard and preparative autosamplers. We also suggest referring to the maintenance and troubleshooting sections of your instrument manual. The best way to avoid getting bubbles in the flow path is to pump the mobile phase through a degasser module.
Lc chromatography troubleshooting hplc columns in india. Abnormal pressure a change in the operating pressure is a sign that there may be a problem. Runser, maintaining and troubleshooting hplc systems a users guide, wiley, ny 1981. Use only hplc grade solvents, salts, ion pair reagents, and base and acid modifiers. Gradient elution overcomes this problem by decreasing the retention of. Pumps for hplc and uhplc deliver mobile phase through the column that contains the stationary phase. Page 2 trouble shooting steps you have recognized there is a problem. The 1220 infinity ii lc delivers high quality for an affordable price. This guide is intended as an aid in troubleshooting problems that occur in the day to day running of your. For more information, please visit us at agilent 1200. Final tips and tricks hplc troubleshooting agilent. Agilent technologies makes no warranty of any kind with regard to this material. 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. Manual for the agilent 1100 series vacuum degasser.
Thats why we created maintaining your agilent 1100 series hplc system. Hplc questions and answers pdf free download in biochemistry mcqs,interview questions,objective questions,multiple choice. Peak response performance monitoring use your test method known performance inject multiple runs. 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. Agilent 2100 bioanalyzer maintenance and troubleshooting guide. Pdf highperformance liquid chromatography is one of the most used analytical techniques in industry. Baseline troubleshooting column troubleshooting other issues. The important segments of hplc system at the same, whether we use a modular system or a more sophisticated unit. 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. Carryover carryover contamination in hplc and lcms systems. Care, maintenance, and troubleshooting oftroubleshooting. Page 1 tips and tricks of hplc system troubleshooting agilent technologies, inc. Useful data is provided throughout the book, such as. Liquid chromatography lc is a method of column chromatography, performed for the purpose of analyzing mixtures.
Even the improvements added in later time were able to shorten the analysis time slightly. Pumps for hplc and uhplc deliver mobile phase through the column that. Hplc troubleshooting hints and tips chromatography. 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. Agilent technologies shall not be liable for errors contained herein or for incidental, or consequential damages in connection with the furnishing. If the pressure is normal, there is a problem with the column. Report data files for the equivalent signal data files. The infinitylab analytical hplc systems are highly efficient hplc and uhplc systems that feature the latest technology. Injector and injection solvents the injector rapidly introduces the sample into the system with minimal disruption of the solvent flow. Agilent technologies 1200 series user manual pdf download. How do i inhibit bubbles from getting into the flow cell. Standard hplc columns usually contain 5 m silicabased particles. Agilents zorbax column selection guide for hplc is the only chromatography guide that combines a broad offering of lc columns with expert advice that can help you achieve reliable results faster. Using these characters in file or directory names may cause problems when.
The presence of algae in hplc systems can cause a variety of problems that may be incorrectly diagnosed as instrument or application problems. We have 14 agilent technologies 1260 infinity manuals available for free pdf download. If ace does not outperform your existing column of equivalent phase, particle size and dimensions, send in your comparative data within 60 days and keep the ace column free of charge. Hardware that provides rapid instrument stabilization and excellent reproducibility of analytical results, and software that includes analytical workow automation and overlap injection functions and features a userfriendly analysis screen view contribute to.
143 1011 366 227 1193 643 1027 1056 956 61 1082 1443 440 309 411 364 1334 965 486 586 1268 263 1497 1080 1371 522 887 194 834 1509 975 252 1253 1356 688 840 1331 1168 48 354 1304 762 1134 77 692 587 1492 222 735 1285