ACQUITY UPLC H-Class System Guide - Waters …

1 ACQUITY uplc H-ClassSystem GuideRevision BCopyright Waters Corporation 2010 All rights reservediiCopyright notice 2010 Waters CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA AND IN IRELAND. ALL RIGHTS RESERVED. THIS DOCUMENT OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE information in this document is subject to change without notice and should not be construed as a commitment by Waters Corporation. Waters Corporation assumes no responsibility for any errors that may appear in this document. This document is believed to be complete and accurate at the time of publication. In no event shall Waters Corporation be liable for incidental or consequential damages in connection with, or arising from, its , ACQUITY uplc , Connections INSIGHT, Millennium, PIC, uplc , and Waters are registered trademarks, and Auto Blend Plus, eCord, Empower, LAC/E, MassLynx, and THE SCIENCE OF WHAT S POSSIBLE.

2 Are trademarks of Waters is a registered trademark of is a registered trademark of the Shakeproof division of Illinois Tool Works, is a registered trademark of Phillips Screw is a registered trademarks of E. I. du Pont de Nemours and is a registered trademark of Textron is a trademark of Victrex is a trademark of Union Carbide is a trademark of ICI Americas, registered trademarks or trademarks are the sole property of their commentsWaters Technical Communications department invites you to tell us of any errors you encounter in this document or to suggest ideas for otherwise improving it. Please help us better understand what you expect from our documentation so that we can continuously improve its accuracy and seriously consider every customer comment we receive. You can reach us at WatersContact Waters with enhancement requests or technical questions regarding the use, transportation, removal, or disposal of any Waters product.

3 You can reach us via the Internet, telephone, or conventional considerationsSome reagents and samples used with Waters instruments and devices can pose chemical, biological, and radiological hazards. You must know the potentially hazardous effects of all substances you work with. Always follow Waters contact informationContacting mediumInformationInternetThe Waters Web site includes contact information for Waters locations worldwide. Visit and faxFrom the USA or Canada, phone 800 252-HPLC, or fax 508 872 other locations worldwide, phone and fax numbers appear in the Waters Web mailWaters Corporation34 Maple StreetMilford, MA 01757 USAivGood Laboratory Practice, and consult your organization s safety representative for specific to the ACQUITY uplc H-Class instrumentsHigh voltage hazardSafety advisoriesConsult Appendix A for a comprehensive list of warning and caution the ACQUITY uplc H-Class systemWhen operating the ACQUITY uplc H-Class System , follow standard quality-control (QC) procedures and the guidelines presented in this : To avoid electric shock, do not remove the ACQUITY uplc H-Class instrument s protective panels.

4 The components within are not symbolsAudience and purposeThis Guide is intended for personnel who install, operate, and maintain ACQUITY uplc H-Class instruments. It gives an overview of the System s technology and use of the ACQUITY uplc H-Class systemThe Waters ACQUITY uplc H-Class System is for research use calibrate LC systems, follow acceptable calibration methods using at least five standards to generate a standard curve. The concentration range for standards must include the entire range of QC samples, typical specimens, and atypical calibrating mass spectrometers, consult the calibration section of the operator s Guide for the instrument you are calibrating. In cases where an overview and maintenance Guide , not an operator s Guide , accompanies the SymbolDefinitionManufacturerAuthorized representative of the European CommunityConfirms that a manufactured product complies with all applicable European Community directivesAustralia C-Tick EMC CompliantConfirms that a manufactured product complies with all applicable United States and Canadian safety requirementsConsult instructions for useviinstrument, consult the instrument s online Help System for calibration run three QC samples that represent subnormal, normal, and above-normal levels of a compound.

5 Ensure that QC sample results fall within an acceptable range, and evaluate precision from day to day and run to run. Data collected when QC samples are out of range might not be valid. Do not report these data until you are certain that the instrument performs analyzing samples from a complex matrix such as soil, tissue, serum/plasma, whole blood, and other sources, note that the matrix components can adversely affect LC/MS results, enhancing or suppressing ionization. To minimize these matrix effects, Waters recommends you adopt the following measures: Prior to the instrumental analysis, use appropriate sample pretreatment such as protein precipitation, liquid/liquid extraction (LLE), or solid phase extraction (SPE) to remove matrix interferences. Whenever possible, verify method accuracy and precision using matrix-matched calibrators and QC samples. Use one or more internal standard compounds, preferably isotopically labeled classificationISM Classification: ISM Group 1 class BThis classification has been assigned in accordance with CISPR 11 Industrial Scientific and Medical (ISM) instruments requirements.

6 Group 1 products apply to intentionally generated and/or used conductively coupled radio-frequency energy that is necessary for the internal functioning of the equipment. class B products are suitable for use in both commercial and residential locations and can be directly connected to a low voltage, power-supply authorized representativeWaters Corporation (Micromass UK Ltd.)Floats RoadWythenshaweManchester M23 9 LZUnited KingdomTelephone:+44-161-946-2400 Fax:+44-161-946-2480 Contact:Quality managerviiiTable of ContentsixCopyright notice .. iiTrademarks .. iiCustomer comments .. iiiContacting Waters .. iiiSafety considerations .. iii Considerations specific to the ACQUITY uplc H-Class instruments .. iv Safety advisories .. ivOperating the ACQUITY uplc H-Class System .. iv Applicable symbols .. v Audience and v Intended use of the ACQUITY uplc H-Class v Calibrating .. v Quality-control .. viISM classification.

7 Vi ISM Classification: ISM Group 1 class B .. viEC authorized representative .. vii1 ACQUITY uplc H-Class System .. 1-1 UltraPerformance Liquid Chromatography .. 1-2 Features of the ACQUITY uplc H-Class System .. 1-4 Flow-through-needle injector .. 1-4 Wash solvent .. 1-4 Purge 1-5 Active solvent conditioning .. 1-5 Software enhancements .. 1-5 System components .. 1-7 Column technology .. 1-9 For additional information .. 1-11 Table of ContentsxTable of Contents2 Optimizing Performance .. 2-1 General guidelines .. 2-2 Dispersion .. 2-3 Carryover .. 2-4 Reproducibility .. 2-5 Cycle time (between injections) .. 2-5 Preventing leaks .. 2-5 Sample preparation .. 2-5 Particulates .. 2-6 Matching sample diluents .. 2-6 Transferring methods .. 2-6 Measuring dwell volume .. 2-8 Transferring from HPLC to uplc .. 2-9 Transferring from uplc to HPLC .. 2-10 Developing methods.

8 2-113 Preparing the System .. 3-1 Preparing System hardware .. 3-2 Powering-on the System .. 3-2 Monitoring startup 3-2 Monitoring System module LEDs .. 3-3 Power LED .. 3-3 Status LEDs .. 3-3 Enabling the leak sensors .. 3-5 Starting up the System .. 3-5 Configuring chromatography data software .. 3-8 ACQUITY control panels .. 3-8 Quaternary solvent manager control 3-8 Sample manager control panel .. 3-11 TUV detector control panel .. 3-12 Column manager control panel .. 3-14 Table of ContentsxiStarting the ACQUITY uplc Console .. 3-15A Safety Advisories .. A-1 Warning symbols .. A-2 Task-specific hazard A-2 Specific warnings .. A-3 Caution symbol .. A-5 Warnings that apply to all Waters instruments .. A-6 Electrical and handling symbols .. A-11 Electrical symbols .. A-11 Handling symbols .. A-12B External Connections .. B-1 System tubing connections.

9 B-1 Instrument external wiring connections .. B-3 ACQUITY uplc H-Class instrument external wiring B-3 Ethernet connections .. B-4 Column heater B-4 Signal connections .. B-4 Making signal connections .. B-4 Connecting to the electricity source .. B-9C Solvent Considerations .. C-1 Introduction .. C-2 Preventing contamination .. C-2 Clean solvents .. C-2 Solvent quality .. C-2 Solvent preparation .. C-2 Water .. C-2 Solvent recommendations .. C-3 General solvent guidelines .. C-4 Solvents to avoid .. C-6xiiTable of Contents ACQUITY uplc H-Class System recommendations .. C-6 Quaternary solvent manager C-7 Sample manager C-8 Detector C-8 Common solvent properties .. C-8 Solvent miscibility .. C-10 Using miscibility numbers (M-numbers).. C-11 Solvent stabilizers .. C-12 Solvent viscosity .. C-12 Wavelength selection .. C-12 UV cutoffs for common solvents.

10 C-12 Mixed mobile C-13 Mobile phase C-141-11 ACQUITY uplc H-Class SystemWaters designed the ACQUITY uplc H-Class System to perform both uplc and HPLC analyses. Hardware and software features facilitate transferring between the two types of chromatography and developing new Liquid Chromatography1-2 Features of the ACQUITY uplc H-Class system1-4 System components1-71-2 ACQUITY uplc H-Class SystemUltraPerformance Liquid ChromatographyIn 2004, Waters made significant advances in instrumentation and column design to introduce uplc technology to the field of separation science. By employing this technology, Waters ACQUITY uplc systems achieve a marked increase in resolution, speed and sensitivity in liquid chromatography when compared to conventional liquid chromatography uses columns packed with m-diameter, round particles and operating pressures up to 15,000 psi. The van Deemter equation, an empirical formula that describes the relationship between linear velocity and column efficiency, considers particle size as one of the variables.