Waters ACQUITY, System Manual

Page 1: ...ACQUITY UPLC H Class System Guide 715005049 Revision C Copyright Waters Corporation 2016 All rights reserved...

Page 2: ...its use For the most recent revision of this document consult the Waters Web site waters com Trademarks ACQUITY is a registered trademark of Waters Corporation ACQUITY UPLC is a registered trademark o...

Page 3: ...to report any errors that you encounter in this document or to suggest ideas for otherwise improving it Help us better understand what you expect from our documentation so that we can continuously im...

Page 4: ...consulted in all cases where the symbol is used to find out the nature of the potential hazard and any actions which have to be taken Power cord replacement hazard Warning To avoid electric shock use...

Page 5: ...5 of the FCC Rules Operation is subject to the following two conditions 1 this device may not cause harmful interference and 2 this device must accept any interference received including interference...

Page 6: ...nts Consult instructions for use Alternating current Electrical and electronic equipment with this symbol may contain hazardous substances and should not be disposed of as general waste For compliance...

Page 7: ...where an overview and maintenance guide not an operator s guide accompanies the instrument consult the instrument s online Help system for calibration instructions Quality control Routinely run three...

Page 8: ...adio 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 t...

Page 9: ...Equipment misuse notice v Safety advisories v Operating the system v Applicable symbols vi Audience and purpose vi Intended use vii Calibrating vii Quality control vii EMC considerations vii Canada sp...

Page 10: ...2 4 Cycle time between injections 29 2 5 Preventing leaks 29 2 5 1 Installation recommendations for fittings 30 2 6 Developing methods 41 2 7 Sample preparation 41 2 7 1 Particulates 41 2 7 2 Matchin...

Page 11: ...maintenance 63 4 1 Contacting Waters Technical Service 63 4 1 1 Viewing module information 63 4 2 Maintenance procedures and frequency 64 4 3 Spare parts 64 4 4 Troubleshooting with Connections INSIGH...

Page 12: ...ectrical symbols 84 A 8 Handling symbols 85 B Solvent considerations 86 B 1 Solvent recommendations 86 B 2 Recommended solvents 87 B 3 Preventing contamination 87 B 3 1 Clean solvents 87 B 3 2 Solvent...

Page 13: ...cations 102 C 1 2 Environmental specifications 103 C 1 3 Electrical specifications 103 C 1 4 Physical Specifications 104 C 2 Performance specifications 104 C 2 1 QSM and bioQSM performance specificati...

Page 14: ...y In 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...

Page 15: ...ure above that using 1 7 m particles achieves higher efficiency that persists as flow rate increases lower HETP indicates higher efficiency When operating in this area of the plot the peak capacity an...

Page 16: ...parations were identical except for the flow rate which was scaled based on particle size 1 2 Features of the ACQUITY UPLC H Class system The ACQUITY UPLC H Class system combines the speed and perform...

Page 17: ...ks are completed the solvent manager begins to deliver the gradient to the injection valve The dwell volume of the system which affects the amount of time required for this gradient to reach the colum...

Page 18: ...he flow ramping should be adjusted 1 2 1 7 Auto Blend Plus Auto Blend Plus technology uses pure solvents and concentrated stocks to blend mobile phase compositions at a specific pH At the same time it...

Page 19: ...manager sample manager column heater detector tunable ultraviolet photodiode array evaporative light scattering fluorescent conductivity refractive index or mass spectrometry and an ACQUITY UPLC colu...

Page 20: ...aration The exterior of the needle is washed in the injection port as specified in the method The standard injection volume range of the SM FTN is 10 L however this can be expanded up to 1 mL through...

Page 21: ...s not suitable for active preheating 1 3 4 Column manager optional The ACQUITY UPLC H Class column manager is an option for helping to ensure precise reproducible separations The column manager can re...

Page 22: ...sample organizer you use with your system is compatible with sample managers that have a rotary tray The sample organizer stores multiple microtiter or vial plates and transfers them to and from the...

Page 23: ...ith dual detection split stack configurations 1 3 9 Column technology The ACQUITY UPLC columns are packed with the following 1 7 m bridged ethylsiloxane hybrid 1 8 m high strength silica particles or...

Page 24: ...Guide ACQUITY UPLC Sample Manager Flow Through Needle Overview and Maintenance Guide ACQUITY UPLC Column Heater Active Overview and Maintenance Guide ACQUITY UPLC Column Manager Active and Column Man...

Page 25: ...formation and to join the ACQUITY UPLC online community where you can do these things Share information with and ask questions of ACQUITY UPLC experts and scientists Access ACQUITY UPLC publications a...

Page 26: ...es efficient management of cycle time essential When performing fast UPLC analyses a peak of interest can be less than 0 5 seconds in width Waters recommends a sampling rate that will generate between...

Page 27: ...e Rotate the column or optional in line filter to install or remove it Always use solvent filters on tubing lines in solvent bottles Use the Load Ahead option when you desire a shorter cycle time Prim...

Page 28: ...Small particle chromatography uses small high efficiency columns A typical 2 1 50 mm UPLC column has an approximate 174 L volume compared with 2 5 mL for a typical 4 6 150 mm HPLC column The smaller c...

Page 29: ...he time between analyses The sample manager has a load ahead option that can help decrease cycle time This option instructs the sample manager to aspirate the next sample while the current sample is r...

Page 30: ...re that the tubing is fully bottomed in the fitting port before tightening the compression screw For easier accessibility use long compression screws to attach tubes to the injector and vent valve Per...

Page 31: ...ssion screw at the 12 o clock position 6 Mark the fitting body at the 9 o clock position 7 Ensure that the tubing makes contact with the bottom of the fitting body and use the 1 4 inch open end wrench...

Page 32: ...talled on 1 8 inch OD tubing First use or re installed Compression screw Lock ring Ferrule End of lock ring with larger inside diameter ID Tighten the fitting finger tight 2 5 1 5 Short 1 4 28 fitting...

Page 33: ...with filter and stainless steel lock ring First use or re installed Compression screw Lock ring Ferrule and filter End of lock ring with larger inside diameter ID Tighten the fitting finger tight 2 5...

Page 34: ...fittings see Assembling new fittings Tip To prevent band spreading ensure that the tubing is fully bottomed in the fitting before tightening the compression screw First use tightening Re installed Lo...

Page 35: ...steel ferrule Tighten the fitting finger tight plus an additional 3 4 turn using a 1 4 inch open end wrench For detailed instructions about assembling new fittings see Assembling new fittings Tip To...

Page 36: ...ight plus as much as an additional 1 6 turn using a 1 4 inch open end wrench Re installed tightening 2 5 1 9 PEEK fitting with PEEK ferrule and stainless steel lock ring First use or re installed Comp...

Page 37: ...collet First use Hole for inserting the collet and compression screw multi tool Compression screw Collet PEEK ferrule To tighten this fitting for first use 1 Finger tighten the fitting 2 Using the col...

Page 38: ...llet and compression screw multi tool Column or column in line filter 5 16 inch open end wrench Tip To prevent band spreading ensure that the tubing bottoms in its fitting before you tighten the compr...

Page 39: ...f the fitting Re installed Hole for collet and compression screw multi tool Collet and compression screw multi tool Collet PEEK ferrule Compression screw 2 Tighten the fitting finger tight 3 Using the...

Page 40: ...inch open end wrench Tip When reinstalling the fitting examine the collet and ferrule for damage and replace them if necessary always loosen the collet before re connecting the reusable finger tight...

Page 41: ...for Ion Exchange Size Exclusion and Reversed phase Chromatography documentation For the greatest flexibility in method development Waters recommends configuring the system with the column manager and...

Page 42: ...nsfer are the ideal tools to achieve that goal with quaternary solvent capability low pressure mixing and flow through needle design The optional column manager facilitates switching between target co...

Page 43: ...column dimensions dwell volumes configurations injection volumes analyte molecular weights and gradient profiles You must calculate various measurements of both systems using the same method to ensure...

Page 44: ...ACQUITY UPLC H Class system An optional passive column stabilizer is available for existing chromatographic methods not suitable for active preheating Select the column with the most similar selectivi...

Page 45: ...a gradient hold is required Identical chemistries are available in UPLC and HPLC columns simplifying their selection and allowing for a simple method transfer from the ACQUITY UPLC H Class system to...

Page 46: ...owing start up tests run CPU board memory RAM and ROM external communication system Ethernet and clock If the start up tests indicate a malfunction consult the console online Help Important If the sys...

Page 47: ...perform these tasks 3 2 1 To open the console from Empower software 1 From the Empower navigation bar select Run Samples and then Control Panel Result A control panel for each device in the system app...

Page 48: ...nsole you can select the Startup System feature to prime all solvents and to specify the solvent composition flow rate column and sample temperatures and needle characterization For details see the co...

Page 49: ...ilure is preventing operation Cycle power to the sample manager If the LED remains red report the problem to Waters Technical Support Alternative Firmware upload is complete 3 4 2 2 Flow LED Flow stat...

Page 50: ...tion Unlit If the detector is equipped with a lamp the lamp is extinguished If the detector is not equipped with a lamp the detector is idle Steady green If the detector is equipped with a lamp the la...

Page 51: ...uxiliary CM AUX Other compatible column modules are controlled via the sample manager control panel If Empower software controls the system the column manager s control panel appears at the bottom of...

Page 52: ...et points while the system is running samples Figure 3 2 SM FTN control panel Run LED Mirrors the run status LED on the sample manager s front panel unless communications are interrupted Current sampl...

Page 53: ...edit these parameters when the system is idle by clicking the underlined value You cannot edit solvent manager parameters while the system is running samples Figure 3 3 QSM control panel Flow LED Mirr...

Page 54: ...ELS control panel The evaporative light scattering ELS detector s control panel displays light scattering units photomultiplier tube gain factor gas pressure nebulizer temperature and drift tube tempe...

Page 55: ...ntrol panel function Description Autozero Resets the detector offsets Reset module Resets the detector after an error condition Help Displays the console online Help 3 5 5 FLR control panel The fluore...

Page 56: ...nal functions by right clicking anywhere in the detector control panel Table 3 8 Additional functions in the FLR detector control panel Control panel function Description Autozero Resets the detector...

Page 57: ...blocking Status Displays the status of the current operation You can access these additional functions by right clicking anywhere in the detector control panel Table 3 9 Additional functions in the PD...

Page 58: ...is negative the chromatogram is inverted Signal measurement Displays the signal in RIU Status Displays the status of the current operation You can access these additional functions by right clicking...

Page 59: ...gth mode the value of wavelength B also appears AU Displays the absorbance units of wavelength A If the detector is in dual wavelength mode the absorbance units of wavelength B also appears Status Dis...

Page 60: ...e the Start up system function to prime the solvent manager after changing the mobile phase after changing the sample needle or after the system has been idle a long period of time for example overnig...

Page 61: ...Prime Solvents tab click the sample manager sub tab and if necessary change the settings for the wash and purge solvent Table 3 12 Sample manager priming parameter values Parameter Wash solvent Purge...

Page 62: ...do not power on operate or ignite the lamp of the detector when there is no flow through the cell or when the cell is dry Note Change the settings on the Optional Characterize tab only if the needle h...

Page 63: ...sage if any Nature of the symptom Serial number of the system module and its firmware version if applicable Flow rate Operating pressure Solvent s Detector settings sensitivity and wavelength Type and...

Page 64: ...Spare parts To ensure that your system operates as designed use only Waters Quality Parts Visit www waters com wqp for information about Waters Quality Parts including how to order them 4 4 Troublesh...

Page 65: ...profile Result A ZIP file containing your Connections INSIGHT profile is forwarded to Waters customer support for review Note Saving a service profile or plot file from the instrument console can requ...

Page 66: ...m service request recommended cleaning flushing and packaging procedures from Waters Technical Service 5 1 Ethernet connections The sample manager incorporates an internal Ethernet switch that accommo...

Page 67: ...tions Figure 5 1 System rear panel cable connections Detector Column heater Sample manager SM FTN Solvent manager QSM Ethernet switch Workstation Interconnect D sub cables Ethernet cables August 8 201...

Page 68: ...3 Plumbing connections Figure 5 2 System plumbing connections Detector Column heater Sample manager SM FTN Solvent manager QSM Stainless steel tubing PEEK tubing August 8 2016 715005049 Rev C Page 68...

Page 69: ...5 4 Waste tubing connections Figure 5 3 System waste tubing connections Solvent bottle tray Detector August 8 2016 715005049 Rev C Page 69...

Page 70: ...interconnect cable 5 5 1 Connecting to a wall electricity source Warning To avoid electrical shock observe these precautions Use SVT type power cord in the United States and HAR type power cord or bet...

Page 71: ...ability To connect to a cart s electricity source 1 Connect the female end of the cart s electrical cables included in the startup kit to the receptacle on the rear panel of each system module 2 Conne...

Page 72: ...gnal cable to the connector Figure 5 5 Positive and negative lead connections Screw Connector Signal cable 3 Fit the grounding cable s fork terminal on the rear panel grounding stud and secure the ter...

Page 73: ...es are compatible with the ACQUITY UPLC H Class system Column heater Column heater 30 cm Column heater cooler Column manager Column manager auxiliary The sample manager powers and communicates with th...

Page 74: ...and column heater cooler before connecting or disconnecting the interconnect cable 1 Make sure the sample manager and the column module are powered off 2 Connect the interconnect cable to the High Den...

Page 75: ...or property damage resulting from the failure of individuals to comply with any safety precaution when installing repairing or operating any of its instruments or devices The following symbols warn o...

Page 76: ...ents and devices fitted with nonmetallic tubing Warning To avoid injury from bursting nonmetallic tubing heed these precautions when working in the vicinity of such tubing when it is pressurized Wear...

Page 77: ...an process biohazards corrosive materials or toxic materials Warning To avoid personal contamination with biohazards toxic materials or corrosive materials you must understand the hazards associated w...

Page 78: ...nges or modifications to this unit not expressly approved by the party responsible for compliance could void the user s authority to operate the equipment Avertissement Toute modification sur cette un...

Page 79: ...of the tubing Avertissement Manipulez les tubes en polym re sous pression avec precaution Portez syst matiquement des lunettes de protection lorsque vous vous trouvez proximit de tubes en polym re pre...

Page 80: ...costante Non utilizzare tubi eccessivamente logorati o piegati Non utilizzare tubi non metallici con tetraidrofurano THF o acido solforico o nitrico concentrati Tenere presente che il cloruro di metil...

Page 81: ...THF Methylene chloride Dimethyl sulfoxide THF This warning applies to Waters instruments fitted with nonmetallic tubing This warning applies to instruments operated with flammable solvents August 8 2...

Page 82: ...recchiatura secondo modalit non previste dal produttore pu compromettere la protezione offerta dall apparecchiatura Advertencia el usuario deber saber que si el equipo se utiliza de forma distinta a l...

Page 83: ...ion does not appear on the instrument or device Warning To protect against fire replace fuses with those of the type and rating indicated in the Replacing fuses section of the Maintenance Procedures c...

Page 84: ...strument manuals and on an instrument s front or rear panels Symbol Description Electrical power on Electrical power off Standby Direct current Alternating current Alternating current 3 phase Safety g...

Page 85: ...can appear on labels affixed to the packaging in which instruments devices and component parts are shipped Symbol Description Keep upright Keep dry Fragile Use no hooks Upper limit of temperature Lowe...

Page 86: ...TY UPLC FLR detector 2432 Conductivity detector B 1 Solvent recommendations The system is designed for reversed phase chromatography and ACQUITY UPLC BEH column chemistries Waters evaluated the system...

Page 87: ...rift and they can clog solvent reservoir filters inlet filters and capillary lines B 3 2 Solvent quality Use MS grade solvents for the best possible results the minimum requirement is HPLC grade Filte...

Page 88: ...h a 20 methanol water solution to prevent microbial growth Notice Some buffers can be incompatible with mass spectrometers Consult the documentation that accompanies your instrument for compatible buf...

Page 89: ...stems with hexane or THF at high concentrations and high pressure and is recommended for many ELS detector based applications where THF is used in the mobile phase at high concentrations B 7 Additives...

Page 90: ...THF dioxane and diisopropylether If you must use peroxidizable compounds be sure to filter them through dry aluminium oxide to adsorb formed peroxides Solutions that contain strong concentrations of...

Page 91: ...TY UPLC H Class systems Nevertheless you can use these solvents in weak dilutions 5 as additives sample diluents or modifiers Contact your Waters sales representative or local technical support organi...

Page 92: ...n cause salt build up on the needle and wash port which can require periodic cleaning B 14 Common solvent properties The following table lists the properties for some common chromatographic solvents T...

Page 93: ...20 C 107 7 28 Isopropyl alcohol 32 4 at 20 C 82 26 12 Isopropyl myristate 1 at 20 C 182 6 164 Methanol 97 at 20 C 64 7 11 Methyl t butyl ether 240 at 20 C 55 2 28 Methyl ethyl ketone 74 at 20 C 79 64...

Page 94: ...add the organic solvent Note cutoff is the wavelength at which the absorbance of the solvent equals 1 AU Table B 2 Solvent miscibility Polarity index Solvent Viscosity cP 20 C at 1 atm Boiling point C...

Page 95: ...le with each other A liquid is classified in the M number system by testing for miscibility with a sequence of standard solvents A correction term of 15 units is then either added or subtracted from t...

Page 96: ...ne 275 Ethanol 210 Isopropanol 205 Isopropyl ether 220 Methanol 205 n Propanol 210 Tetrahydrofuran THF 230 B 14 4 2 Mixed mobile phases The following table provides approximate wavelength cutoffs for...

Page 97: ...ic acid 0 1 190 Morpholinoethanesulfonic acid MES 10 mM pH 6 0 215 Potassium phosphate monobasic 10 mM 190 Potassium phosphate dibasic 10 mM 190 Sodium acetate 10 mM 205 Sodium chloride 1 M 207 Sodium...

Page 98: ...h Table B 5 Mobile phase absorbance measured against air or water Absorbance AU at specified wavelength nm 200 205 210 215 220 230 240 250 260 280 Solvents Acetonitrile 0 05 0 03 0 02 0 01 0 01 0 01 M...

Page 99: ...0 90 0 45 0 06 0 01 Potassium phosphate monobasic KH2PO4 10 mM 0 03 0 01 Potassium phosphate dibasic K2HPO4 10 mM 0 53 0 16 0 05 0 01 0 01 Sodium acetate 10 mM 1 85 0 96 0 52 0 30 0 15 0 03 0 01 Sodi...

Page 100: ...0 06 0 03 0 02 0 02 0 02 0 01 0 01 3 3 cholamidopropyl dimethylammonio 1 propanesulfonate CHAPS 0 1 2 40 2 32 1 48 0 80 0 40 0 08 0 04 0 02 0 02 0 01 Sodiumdodecyl sulfate SDS 0 1 0 02 0 01 0 01 4 oc...

Page 101: ...line Leak detection Leak sensors installed in drip trays Quantum sychronization Injection synchronization between pump and sample manager enhances retention time reproducibility Settable flow rate ra...

Page 102: ...100 mL mixer filter Optional stainless steel 250 mL mixer filter Plunger wash feature Wash pump plungers using seal wash solvent can be primed manually or run automatically No flow shutdown feature A...

Page 103: ...ttribute Specification Acoustic noise system 65 dBA Ambient operating temperature 4 0 to 40 0 C Ambient operating humidity 20 to 80 non condensing C 1 3 Electrical specifications Attribute Specificati...

Page 104: ...a reduction of dielectric strength or surface resistivity Degree 2 refers only to normally nonconductive pollution Occasionally however expect a temporary conductivity caused by condensation d Moistur...

Page 105: ...solvent A with i2Valve Back pressure 4137 to 6895 kPa 41 to 69 bar 600 to 1000 psi with degassed water Flow precision 0 075 RSD or 0 020 min SD whichever is greater based on six replicates i2Valve Te...

Page 106: ...5 C 0 3 C Detector UV 254 nm wavelength Compressibility compensation Automatic and continuous Priming Wet priming can run at flow rates up to 4 mL min Pump seal wash Equipped with a wash system to flu...

Page 107: ...s Chromatography isocratic Mobile phase 10 90 acetonitrile water Flow rate 0 6 mL min Needle volume 1 to 70 Sample mix caffeine 0 03 mg mL 0 2 to 10 0 L with 15 L needle installed no extension loop Co...

Page 108: ...erance range of between 2 0 and 4 0 C At a set point of 4 C with ambient temperature 23 C and humidity 80 maintains a sample temperature of 2 to 8 C At ambient temperatures 23 C and humidity 80 mainta...

Page 109: ...mL standard and 4 mg mL challenge in 10 90 acetonitrile water compared with blanks of 10 90 acetonitrile water Injection volume 5 L Column temperature 40 C Detection UV at 273 nm sampling rate 20 poi...

Page 110: ...onitrile water 0 1 formic acid compared with blanks of 10 90 acetonitrile water 0 1 formic acid Injection volume 5 L Column ACQUITY UPLC BEH C18 1 7 m 2 1 50 mm Column temperature 40 C Sample temperat...

Page 111: ...Minimum sample compartment temperature specifications 0 5 10 15 20 25 30 4 8 12 16 20 24 28 32 36 40 Ambient temperature C Maximum compartment temperature Minimum compartment temperature Minimum susta...

Page 112: ...t for extended configuration only Single exit drain manages leaks to waste Door open to heater cutoff delay 1 minute maximum Column tracking eCord technology column information management tracks and a...

Page 113: ...ments Controllable temperature range Ambient 15 C to 65 0 C Temperature accuracy Tested to 0 8 C between 20 and 50 C 1 0 C for remaining range Test qualification The temperature measured by a NIST tra...

Page 114: ...column installed No flow Measurement taken with internal temperature sensor Measurement taken after 1 hour of thermal equilibration at set point Door is opened for 30 seconds Tested at 35 C 55 C and 8...

Page 115: ...leak sensor installed under compartment Single exit drain manages leaks to waste Column tracking eCord technology column information management tracks and archives column usage history a Operating wi...

Page 116: ...the sample manager and sample organizer can maintain an average sample temperature of 18 C below ambient 3 0 C Temperature accuracy No more than 1 0 C in temperature between a traceable external tempe...