Waters ACQUITY UPLC H-Class Series System Manual Download Page 44

Figure 2–11: Method transfer from HPLC to UPLC

When transferring methods from one system to another, you must define and characterize the
original method well. Doing so includes noting information about 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 a
successful transfer. Using the Waters ACQUITY UPLC Columns Calculator ensures the best
results for transferring the LC method from HPLC to UPLC or UPLC to HPLC.

April 4, 2018, 715005702 Rev. A

Page 44

Summary of Contents for ACQUITY UPLC H-Class Series

Page 1: ...ACQUITY UPLC H Class Series System Guide 715005702 Revision A Copyright Waters Corporation 2018 All rights reserved...

Page 2: ...recent revision of this document consult the Waters website waters com Trademarks ACQUITY is a registered trademark of Waters Corporation ACQUITY UPLC is a registered trademark of Waters Corporation A...

Page 3: ...at 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 improve its accuracy and u...

Page 4: ...ent tray and rails higher than one meter 39 4 inches above the bench top Warning To avoid spinal and muscular injury do not attempt to lift a system module without assistance Warning To avoid crushing...

Page 5: ...nce could void the user s authority to operate the equipment This device complies with Part 15 of the FCC Rules Operation is subject to the following two conditions 1 this device may not cause harmful...

Page 6: ...pean Community directives or Australia EMC compliant Confirms that a manufactured product complies with all applicable United States and Canadian safety requirements Consult instructions for use Alter...

Page 7: ...must include the entire range of QC samples typical specimens and atypical specimens When calibrating mass spectrometers consult the calibration section of the operator s guide for the instrument you...

Page 8: ...ments 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 sui...

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

Page 10: ...UPLC analysis 26 2 2 Dispersion 27 2 3 Carryover 27 2 4 Cycle time between injections 28 2 5 Preventing leaks 28 2 5 1 Installation recommendations for fittings 29 2 6 Developing methods 42 2 7 Sample...

Page 11: ...ontrol panel 59 3 6 7 PDA control panel 60 3 6 8 RI control panel 61 3 6 9 TUV control panel 62 3 6 10 ACQUITY QDa 63 3 6 11 2432 conductivity control panel 64 3 7 Starting up the system 64 4 System m...

Page 12: ...ing symbols 80 B 1 1 Specific warnings 81 B 2 Notices 82 B 3 Bottles Prohibited symbol 83 B 4 Required protection 83 B 5 Warnings that apply to all Waters instruments and devices 83 B 6 Warnings that...

Page 13: ...echnology 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...

Page 14: ...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 15: ...ons were identical except for the flow rate which was scaled based on particle size 1 2 Features of the ACQUITY UPLC H Class Series system The ACQUITY UPLC H Class Series system combines the speed and...

Page 16: ...asks 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 col...

Page 17: ...uld 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 controls the conce...

Page 18: ...a solvent manager sample manager column heater detector tunable ultraviolet photodiode array evaporative light scattering fluorescent conductivity refractive index or mass spectrometry and an ACQUITY...

Page 19: ...needle is part of the fluidic path and the internal surfaces are flushed by the mobile phase during the separation The exterior of the needle is washed in the injection port as specified in the method...

Page 20: ...suitable for active preheating 1 3 4 Column manager optional The ACQUITY UPLC H Class Series column manager is an option for helping to ensure precise reproducible separations The column manager can...

Page 21: ...s via the SM FTN console The switch box is mounted to the rear of the SM FTN The SM FTN interconnect cable is connected to the sample manager SM port of the switch box the CH A interconnect cable is c...

Page 22: ...ion caused by injections of high organic sample diluents For more information on installing this extension loop see Post Injection Volume Kit Instructions 1 3 9 Detection The detectors compatible with...

Page 23: ...n The name of the sample set or sample list run on the column Number of injections onto the column Number of samples injected onto the column The highest pressure that the column has experienced and t...

Page 24: ...tive Light Scattering Detector Getting Started Guide ACQUITY Refractive Index Detector Overview and Maintenance Guide ACQUITY QDa Detector Overview and Maintenance Guide 2432 Conductivity Cond Detecto...

Page 25: ...also makes 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...

Page 26: ...stall 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 Prime properly Prime solvent lines during system start u...

Page 27: ...tography 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 column and particle si...

Page 28: ...uate the valve if you suspect that sample residue in the valve is causing carryover problems Test your sample in the strong wash solvent to make sure that the strong wash solvent does not cause either...

Page 29: ...presentative for your organization regarding its protocols for handling such materials Warning To avoid personal contamination with biologically hazardous materials wear clean chemical resistant powde...

Page 30: ...hreaded fitting Permanent marker To assemble the new fittings 1 Insert the end of a tube into the hexagonal end of the compression screw 2 Insert the tube into the larger end of the ferrule 3 Insert t...

Page 31: ...hten 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 prevent band sprea...

Page 32: ...ece stainless steel ferrule V detail First use Short flats Compression screw 2 piece stainless steel ferrule Tighten the fitting finger tight plus an additional 3 4 turn using a 1 4 inch open end wren...

Page 33: ...ed Short flats Compression screw 2 piece stainless steel ferrule Tighten the fitting finger tight plus as much as an additional 1 6 turn using a 1 4 inch open end wrench Reinstalled tightening April 4...

Page 34: ...re installed 1 Compression screw Ferrule Tighten the fitting finger tight 2 5 1 5 1 4 28 flangeless fitting with 2 piece ferrule First use or re installed 1 Compression screw 2 piece ferrule Tighten t...

Page 35: ...ck ring Ferrule End of lock ring with larger inside diameter ID Tighten the fitting finger tight 2 5 1 7 Short 1 4 28 fitting with flangeless ferrule and stainless steel lock ring installed on 1 16 in...

Page 36: ...mpression screw Lock ring Ferrule and filter End of lock ring with larger inside diameter ID Tighten the fitting finger tight 2 5 1 9 PEEK fitting with PEEK ferrule and stainless steel lock ring First...

Page 37: ...ing To prevent band spreading ensure that the tubing is fully bottomed in the connection port before tightening the fitting 2 5 1 11 Dual threaded fitting with locking cap nut Required tools and mater...

Page 38: ...he fitting 1 Loosen the cap nut from the compression fitting 2 Slide the compression fitting together with the ferrule into the inlet of the column or the in line filter Important To prevent band spre...

Page 39: ...placing the ferrule on the column inlet fitting Always loosen the locking cap nut before reconnecting the fitting 2 5 1 12 Replacing the ferrule on the column inlet fitting Replace the column inlet fe...

Page 40: ...re 2 6 Unscrewing cap nut from fitting 1 knurled compression fitting 2 locking cap nut 2 Slide the 1 knurled compression fitting off the tubing Figure 2 7 Sliding fitting off tubing 1 knurled compress...

Page 41: ...ule on the fitting 6 Slide the 1 knurled compression fitting and ferrule onto the tubing Figure 2 9 Sliding fitting and ferrule onto tubing Tubing 1 knurled compression fitting Note If it is a captive...

Page 42: ...will automate the preparation of any pH specific mobile phase from pure solvents for easier method development 2 7 Sample preparation 2 7 1 Particulates Waters recommends filtering all samples with p...

Page 43: ...n target columns while exhibiting low bandspread and maintaining the same temperature profile as the standard column heater The following example of a method transfer shows the related substances test...

Page 44: ...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 ensur...

Page 45: ...ACQUITY UPLC H Class Series system An optional passive column stabilizer is available for existing chromatographic methods not suitable for active preheating Select the column with the most similar s...

Page 46: ...ion and allowing for a simple method transfer from the ACQUITY UPLC H Class Series system to standard analytical HPLC Finally note that the chromatographic conditions provided by the calculator serve...

Page 47: ...llowing start up tests run CPU board memory RAM and ROM external communication system Ethernet and clock If the start up tests indicate a malfunction contact your local Waters representative 2 Power o...

Page 48: ...n how to perform these tasks 3 2 1 To open the console from Empower software 1 From the Empower navigation bar select Run Samples Result A control panel for each device in the system appears 2 In the...

Page 49: ...lect the Startup System feature to prime all solvents and to specify the solvent composition flow rate column and sample temperatures and needle characterization for your next system start up For deta...

Page 50: ...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 51: ...Detector LED color Description 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 e...

Page 52: ...stem Console or data analysis activities You can update a parameter set point directly from a control panel providing sample analysis is not running When the parameter value is underscored and appears...

Page 53: ...s these additional functions by right clicking anywhere in the column manager control panel Table 3 4 Additional functions in the column manager s control panel Control panel function Description Rese...

Page 54: ...re set point for the sample compartment Current sample compartment temperature Displays the current temperature in the column compartment Status Displays the status of the current operation You can ac...

Page 55: ...ager Stop flow When clicked immediately stops all flow from the solvent manager Pressure Delta The difference between the maximum and minimum pressures observed during the previous minute of operation...

Page 56: ...lunger wash sequence which fills and then slowly empties the primary and accumulator chambers with the current solvent composition while performing a high speed volume seal wash This action helps prev...

Page 57: ...isplays the status of the current operation In this representation of the control panel the status is blank You can access these additional functions by right clicking anywhere in the ISM control pane...

Page 58: ...ing units photomultiplier tube gain factor gas pressure nebulizer temperature and drift tube temperature If Empower software controls the system the detector s control panel appears at the bottom of t...

Page 59: ...ription Auto zero Resets the detector offsets Reset module Resets the detector after an error condition Help Displays the console online Help 3 6 6 FLR control panel The fluorescence FLR detector s co...

Page 60: ...nal functions by right clicking anywhere in the detector control panel Table 3 9 Additional functions in the FLR detector control panel Control panel function Description Auto zero Resets the detector...

Page 61: ...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 10 Additional functions in the...

Page 62: ...ty 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 clickin...

Page 63: ...avelength 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 Stat...

Page 64: ...Eliminates the eluent s contribution to conductivity Cell temperature set point Displays the set point for the flow cell temperature Current cell temperature Displays the current flow cell temperature...

Page 65: ...e solvent manager for minimum of 5 minutes If you are using a concentrated acid and a concentrated base it is recommended that the lines are not primed in succession Instead to reduce the risk of salt...

Page 66: ...mn Manager defaults to Column 1 Depends on type of column compartment Sample temperature On Off or 4 0 to 40 0 C Lamp On On or Off Note For light guiding flow cells do not power on operate or ignite t...

Page 67: ...mptom Serial number of the system module and its firmware version if applicable Flow rate Operating pressure Solvent or solvents Detector settings sensitivity and wavelength Type and serial number of...

Page 68: ...overview and maintenance guide on the documentation media for routine maintenance procedures and frequency 4 3 Spare parts To ensure that your system operates as designed use only Waters Quality Parts...

Page 69: ...ou must transport a system component or remove it from service request recommended cleaning flushing and packaging procedures from Waters Technical Service 5 1 Ethernet connections The sample manager...

Page 70: ...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 April 4 2018...

Page 71: ...5 3 Plumbing connections Figure 5 2 System plumbing connections Detector Column heater Sample manager SM FTN Solvent manager QSM Stainless steel tubing PEEK tubing April 4 2018 715005702 Rev A Page 71...

Page 72: ...5 4 Waste tubing connections Figure 5 3 System waste tubing connections Solvent bottle tray Detector April 4 2018 715005702 Rev A Page 72...

Page 73: ...necting to a wall electricity source Warning To avoid electric shock observe these precautions Use SVT type power cords in the United States and HAR type power cords or better in Europe For requiremen...

Page 74: ...y maintenance operation on it Connect each module to a common ground Recommendation Use a line conditioner and uninterruptible power supply UPS for optimum long term input voltage stability To connect...

Page 75: ...driver attach the positive and negative leads of the signal cable to the connector Tip Refer to the cable connection label affixed to the rear panel of the module Figure 5 5 Positive and negative lead...

Page 76: ...UITY UPLC H Class Series system Column heater CH A Column heater 30 cm CH 30A Column heater cooler 30 cm CHC Column manager CM A Column manager auxiliary CM Aux The sample manager powers and communica...

Page 77: ...e interconnect cable 1 Ensure that the sample manager and the column module are powered off 2 Connect the interconnect cable to the High Density HD port on the rear of the column module 3 Connect the...

Page 78: ...on loop kit To install or replace a post injection loop 1 Power off the sample manager 2 Open the fluidics compartment door 3 Fully insert the post injection loop tubing into port 6 of the injection v...

Page 79: ...fitting into the post injection loop union and then use the 1 4 inch open end wrench to tighten the fitting 3 4 turn beyond finger tight for a new fitting or 1 6 turn beyond finger tight for existing...

Page 80: ...liability in cases of injury 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...

Page 81: ...ies to Waters instruments 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...

Page 82: ...d chemical hazard warning This warning applies to Waters instruments and devices that can process biohazards corrosive materials or toxic materials Warning To avoid personal contamination with biologi...

Page 83: ...t apply to all Waters instruments and devices When operating this device follow standard quality control procedures and the equipment guidelines in this section Warning Changes or modifications to thi...

Page 84: ...loride and dimethyl sulfoxide cause nonmetallic tubing to swell which greatly reduces the rupture pressure of the tubing Avertissement Manipulez les tubes en polym re sous pression avec precaution Por...

Page 85: ...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 86: ...n 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 April 4 2...

Page 87: ...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 88: ...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 89: ...ent 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 ground...

Page 90: ...s devices and component parts are shipped Symbol Description Keep upright Keep dry Fragile Use no hooks Upper limit of temperature Lower limit of temperature Temperature limitation B 9 Stacking system...

Page 91: ...kward until its rear alignment pin rests in the rear alignment slot on the previously added module Figure B 1 Aligning pins with slots Alignment pins Alignment slots 2 Lower the front of the module th...

Page 92: ...ly added module in the system stack Figure B 2 Aligning feet with indents Feet on underside of module being stacked Indents on top side of previously added module 2 Carefully lower the module so that...

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