Waters ACQUITY UPLC Operator's Manual

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ACQUITY UPLC System

Operator's Guide

71500082502/Revision E

−

2010

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Page 2: Copyright Notice
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 use.
Page 3: Customer Comments
Contacting Waters ® Contact Waters with enhancement requests or technical questions regarding the use, transportation, removal, or disposal of any Waters product. You can reach us via the Internet, telephone, or conventional mail. Waters contact information Contacting medium Information...
Page 4: Safety Considerations
Safety considerations Some 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 Good Laboratory Practice, and consult your organization’s safety representative for guidance.
Page 5: Operating The Acquity Uplc Instruments
This guide is intended for personnel who install, operate, and maintain ACQUITY UPLC instruments. Intended use of the ACQUITY UPLC system Waters designed the ACQUITY UPLC system to isolate, concentrate, separate, detect, and measure individual analytes in solution mixtures for research applications of rapid qualitative analysis, quantitative analysis, and/or micropreparative purification.
Page 6: Quality-Control
Class B products are suitable for use in both commercial and residential locations and can be directly connected to a low voltage, power-supply network. EC Authorized Representative Waters Corporation (Micromass UK Ltd.) Floats Road Wythenshawe Manchester M23 9LZ United Kingdom...
Page 7: Table Of Contents
Safety advisories ....................iv Operating the ACQUITY UPLC instruments ..........v Applicable symbols ....................v Audience and purpose..................v Intended use of the ACQUITY UPLC system ............ v Calibrating ......................v Quality-control ....................vi ISM classification ....................vi ISM Classification: ISM Group 1 Class B ............vi EC Authorized Representative .................
Page 8 High temperature column heater ..............1-8 Column manager ....................1-8 Column heater/cooler ..................1-9 30-cm column heater/cooler ................1-9 Optional sample organizer ................1-9 Detectors ......................1-10 TUV detector ....................1-10 PDA detector ....................1-10 ELS detector....................1-10 FLR detector....................1-11 Median baseline filter..................
Page 9 Unpacking and installing the sample organizer ........2-9 Installing the binary solvent manager ............2-10 Installing the sample manager ..............2-10 Installing the HT column heater ..............2-11 Installing the optional column manager ........... 2-12 Installing the optional column heater/cooler ........... 2-13 Installing the detector ...................
Page 10 Sample manager I/O signal connectors ............2-61 TUV detector I/O signal connectors .............. 2-63 PDA detector signal connectors ..............2-64 ELS detector signal connectors ..............2-64 FLR detector signal connectors..............2-64 Signal connections ..................2-64 Connecting to the electricity source ............2-65 Calibrating the XYZ mechanism using the teach block ......
Page 11 Shutting down the system ................3-40 Shutting down for less than 24 hours............3-40 Shutting down for more than 24 hours............3-41 Running HPLC methods on an ACQUITY UPLC system ....... 3-42 System considerations ................... 3-42 Choosing fittings .................... 3-44 4 Configuring System Software .............
Page 12 Performing the gradient performance test ..........5-10 6 Maintaining the System ................ 6-1 Maintenance schedule ..................6-2 Spare parts ......................6-5 Contacting Waters technical service ............6-6 Locating system serial numbers ..............6-6 Maintenance considerations ................6-7 Safety and handling..................6-7 Proper operating procedures ................
Page 13 Resolving sample manager leak sensor errors........... 6-123 Replacing the sample manager’s leak sensor..........6-128 Replacing the sample needle assembly ............6-131 Calibrating the needle Z-axis ..............6-139 Characterizing the needle seal..............6-140 Characterizing the needle and sample loop volumes......... 6-140 Replacing the puncture needle holder ............
Page 14 7 Troubleshooting ..................7-1 Troubleshooting with Connections Insight ..........7-2 Proper operating procedures ................. 7-3 Contacting Waters technical service............... 7-3 Locating system serial numbers ..............7-4 Solving baseline noise and drift problems ..........7-5 Binary solvent manager troubleshooting ..........7-10 Power LED .....................
Page 15 Task-specific hazard warnings................ A-2 Warnings that apply to particular instruments, instrument components, and sample types....................A-3 Caution symbol ....................A-5 Warnings that apply to all Waters instruments ......... A-5 Electrical and handling symbols ..............A-12 Electrical symbols ..................A-12 Handling symbols ..................A-13...
Page 16 Solvent recommendations ................C-4 General solvent guidelines ................C-4 Solvents to use ....................C-4 Solvents to avoid ....................C-6 ACQUITY UPLC system recommendations........... C-6 Binary solvent manager recommendations............ C-8 Sample manager recommendations..............C-8 Detector recommendations................C-8 Solvent miscibility ................... C-8 Using miscibility numbers (M-numbers)............
Page 17 Mixed mobile phases..................C-11 Mobile phase absorbance................C-12 Index ..................... Index-1 Table of Contents xvii...
Page 18 xviii Table of Contents...
Page 19: System Overview
System Overview This section describes the components and features of the ACQUITY ® UPLC system. Contents Topic Page Instruments, components, and data systems UPLC system guidelines Binary solvent manager Sample manager High temperature column heater Column manager Column heater/cooler 30-cm column heater/cooler Optional sample organizer Detectors 1-10...
Page 20: Instruments, Components, And Data Systems
Instruments, components, and data systems ACQUITY UPLC systems include a binary solvent manager, sample manager, column heater, detectors (tunable ultraviolet, photodiode array, evaporative light scattering, fluorescent, or mass spectrometry), and a specialized ACQUITY UPLC column. Small-particle chemistries as utilized in UPLC system chromatography generate narrow peaks.
Page 21: Examples Of Waters Acquity Uplc Systems
Examples of Waters ACQUITY UPLC systems Single detector system with column manager, no mass spectrometer Solvent tray Sample organizer UPLC detector (optional) (optional) Column manager (optional) Sample manager Binary solvent manager Instruments, components, and data systems...
Page 22: Uplc System Guidelines
Tip: When performing fast analyses, note that a peak of interest can be as narrow as 0.5 second. Waters recommends a sampling rate of 25 to 50 points across the peak, which provides good quantitation and peak representation. Sampling rates faster than 20 points per peak yield higher baseline noise and filter time constants must be adjusted accordingly.
Page 23: Acquity Uplc Columns Calculator
100 µL/min ACQUITY UPLC columns calculator The ACQUITY UPLC columns calculator estimates the plate count (N) of an isocratic separation or the peak capacity (Pc) of a gradient separation based on your current HPLC conditions. It then offers you a choice of one or more ACQUITY UPLC columns that can provide increased resolving power in the same amount of time or similar resolving power in less time.
Page 24: How The Binary Solvent Manager Works
Pressure flow envelope 103,421 62,053 Flow rate (mL/min) How the binary solvent manager works Each of the binary solvent manager’s two independent pump systems, A (on the left-hand side) and B (on the right-hand side), contains two linear-drive actuators (left-hand and right-hand). Each left-hand and right-hand actuator pair comprises a single reciprocating “serial”...
Page 25: Sample Manager
Sample manager The ACQUITY UPLC sample manager injects the samples it draws from microtiter plates or vials onto the chromatographic column. A locating mechanism uses a probe to access sample locations and draw sample from them. In the needle overfill load-ahead mode, the sample manager can perform an injection in approximately 15 seconds.
Page 26: High Temperature Column Heater
The high temperature (HT) column heater is modular and its footprint is identical to that of the sample manager. The column heater’s front compartment can accommodate any Waters column up to 4.6 mm ID and 150 mm long. The column rests in a U-shaped tray that swivels outward to receive the column from either side.
Page 27: Column Heater/Cooler
Column heater/cooler The optional column heater/cooler can maintain four columns in a series from 10 to 90 °C (50 to 194 ºF), but does not have switching valves. One column’s eCord connects to the top port to track column usage. 30-cm column heater/cooler The optional 30-cm column heater/cooler can regulate the temperature of HPLC columns up to 30 cm long, from 4 to 65 °C (39.2 to 149 ºF).
Page 28: Detectors
ELS detector The ACQUITY UPLC ELS detector is an evaporative light scattering detector designed for use in the ACQUITY UPLC system. This detector can be controlled by Empower or MassLynx software. The detector incorporates a flow-type nebulizer that is optimized for ACQUITY UPLC system performance.
Page 29: Flr Detector
TQ, refer to the documentation included with it. SQ detector The SQ detector is a single-quadrupole, atmospheric pressure ionization (API) mass spectrometer. Designed for routine ACQUITY UPLC/MS analyses, it can scan at speeds up to 10,000 Da/s. TQ detector The TQ detector is a tandem quadrupole, atmospheric pressure ionization (API) mass spectrometer.
Page 30: Data Systems
The base version of Empower software supports data from TUV, PDA, ELS, and FLR detectors, and single quadrupole mass spectrometers. Popular software options for ACQUITY UPLC system users include System Suitability, Chemical Structures, and Method Validation Manager. Empower online Help.
Page 31: Ecord Column Chip
ACQUITY UPLC columns include an eCord column chip that tracks the usage history of the column. The eCord column chip interacts with the system software, recording information for up to 50 sample queues run on the column.
Page 32: For Additional Information
For additional information Refer to the following documents for further information: • ACQUITY UPLC Quick Reference Card (part number 71508250006) • ACQUITY UPLC System Bookshelf Documentation CD (part number 71500082521) – ACQUITY UPLC Photodiode Array Detector Getting Started Guide (part number 71500108703) –...
Page 33: System Setup
System Setup Contents Topic Page Before you begin Assembling the FlexCart Unpacking and installing the sample organizer Installing the binary solvent manager 2-10 Installing the sample manager 2-10 Installing the HT column heater 2-11 Installing the optional column manager 2-12 Installing the optional column heater/cooler 2-13 Installing the detector...
Page 34: Before You Begin
±1 degree of ground level. If you discover any damage or discrepancy when you inspect the contents of the cartons, immediately contact the shipping agent and your local Waters representative. If you are located in the USA or Canada, report malfunctions or other problems to Waters Technical Service (800 252-4752).
Page 35: Tools
Tools You will need the following tools and materials to install the Waters ® ACQUITY UPLC system. • 5/16-inch open-end wrench (2) • 5/8-inch open-end wrench • 1/4-inch open-end wrench ® • Phillips screwdriver • Small mirror ® • T25 TORX...
Page 36 FlexCart components Monitor mounting Top tray bracket Power Shelf mounting connectors (2) holes Lock knobs (2) Height adjusting screw TP02496 T25 TORX screws System Setup...
Page 37 Remove the 4 Phillips screws from the support plate at the rear of the monitor, and mount the monitor onto the movable arm assembly. Refer to the IBM instruction book, if necessary. Screws (4) Line cord Video cable Plastic channel Movable arm bracket Loosen the 2 T25 TORX screws, at the bottom left-hand side of the cart,...
Page 38: Adjusting The Flexcart's Height
Insert the keyboard shelf’s 2 captive thumbscrews into slots on the cart’s front panel at a level that affords comfortable and safe operation. Turn the thumbscrews 1/4-turn clockwise to lock the position of the shelf. TP02499 Keyboard shelf thumbscrews Adjusting the FlexCart’s height To adjust the cart’s height Loosen the side lock knobs before raising or lowering the top portion of the cart.
Page 39: Locking The Flexcart In Place
Insert the crank handle into the bottom front of the cart, and turn it to raise or lower the cart. Crank handle If your system includes a mass spectrometer and it is positioned on Tip: the right-hand side of the system stack, set the cart to a height that minimizes the length of tubing needed between the instruments stacked on the cart and the mass spectrometer.
Page 40 To release the FlexCart floor lock brake Release the cart by depressing the brake release bar located at the front of the cart. Release bar Lock pedal System Setup...
Page 41: Moving The Assembled Flexcart
Once it is assembled, you can move the cart to other areas of a lab to minimize tubing runs between the ACQUITY UPLC instruments and a mass spectrometer. Use the lip on top of the cart to pull it.
Page 42: Installing The Binary Solvent Manager
Installing the binary solvent manager To install the binary solvent manager To avoid back injuries, two or more people must lift the Warning: binary solvent manager. Lift the binary solvent manager onto the bench top. Or, if your system includes the optional FlexCart, lift the binary solvent manager onto the cart. If your system includes the optional sample organizer, unscrew the existing 7/8-inch-high feet from the bottom of the binary solvent manager and replace them with the 1/4-inch high feet from the...
Page 43: Installing The Ht Column Heater
PEEK™ fittings, sample syringes, and wash syringes inside the Tip: sample manager may have loosened during shipping. To prevent leaks, ensure all PEEK fittings, sample syringes, and wash syringes are tight. Installing the HT column heater Required materials ® • T10 TORX screwdriver •...
Page 44: Installing The Optional Column Manager
Installing the optional column manager To install the optional column manager Place the column manager on a flat surface. Open the column manager door. To avoid poor chromatographic performance, do not Caution: remove the switching valve fittings. They are preset and tested at the factory.
Page 45: Installing The Optional Column Heater/Cooler
Grasp the tan PEEK outlet tube on the upper left-hand side of the drawer and gently pull it forward until you feel it tighten. Tan PEEK outlet tube Place the column manager atop the sample manager, ensuring that the feet are properly positioned in the indentations of the sample manager. Leave the column manager door open.
Page 46: Installing The 30-Cm Column Heater/Cooler
drip tray over the drain routing hole, on the top left-hand side of the HT column heater, column manager, or column heater/cooler. Indentations for feet placement Drain routing hole for drip management system TP02465 Place the solvent tray atop the detector. Installing the 30-cm column heater/cooler If your system includes a 30-cm column heater/cooler, follow the procedure below to install it.
Page 47 To install the 30-cm column heater/cooler To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to an instrument. To completely interrupt power to an instrument, set the power switch to Off, and then unplug the power cord from the AC outlet. After power is removed, wait 10 seconds thereafter before you disconnect an assembly.
Page 48 Mount the rear tab of the adaptor bracket onto the ground stud at the rear of the sample manager. Bracket Ground stud 2-16 System Setup...
Page 49 If your system includes a sample organizer, position the bracket so that its inset area fits over the sample organizer leg. TP02943 Sample Bracket inset organizer leg Installing the 30-cm column heater/cooler 2-17...
Page 50 Insert 2 M4 × 16 screws into the slots on the bracket’s left-hand panel, leaving the first screw loose until both screws are inserted. Then, tighten both screws. M4 side panel spacers with cover screws Ground stud M4 × 16 screws ®...
Page 51 Insert the 3 M4 blue cover screws through the 3 side panel spacers, and then screw them into the 3 threaded holes in the bracket’s center panel. TP02939 Spacer Cover screw Mount the 30-cm column heater/cooler onto the bracket, fitting the column heater/cooler mounting holes over the M4 cover screws.
Page 52: Plumbing The System
Using a 1/4-inch bit driver, attach the black tubing clips to the front of the bracket. Load the clip into the bit driver in the orientation desired, and then snap into the bracket. Tubing clip 1/4-inch bit driver TP02945 TP02948 Plumbing the system To prevent contamination, wear clean, chemical-resistant, Caution:...
Page 53: Installation Recommendations For Fittings
• For easier accessibility, use long compression screws to attach tubes to the injector and vent valve. When tightening system fittings, consult the following table. Installation recommendations for ACQUITY UPLC fittings Fitting Recommended tightening 1/4-28 flangeless with ferrule Snug plus 1/4-turn 10-32 LT135 PEEK with ferrule Snug plus 1/4-turn;...
Page 54: Plumbing A Tuv Detector
Installation recommendations for ACQUITY UPLC fittings (Continued) Fitting Recommended tightening Stainless steel with 2-piece stainless Finger-tight plus 3/4-turn with steel ferrule (first use) wrench Stainless steel with 2-piece stainless Finger-tight plus 1/4-turn with steel ferrule (re-installed) wrench Reusable finger-tight (first use)
Page 55 maintains a minimum post-column pressure of 1724 kPa (17 bar, 250 psi), eliminating post-column outgassing and ensuring a smooth baseline. When the backpressure regulator is installed, the system maintains at Tip: least 1724 kPa (17 bar, 250 psi) backpressure, regardless of the outlet tubing configuration and flow rate, provided that there is some positive flow.
Page 56 Do not install the backpressure regulator if you are connecting to a Rule: second detector or mass spectrometer. Backpressure regulator From detector outlet To waste To avoid spills, empty the waste container at regular Warning: intervals. Route the long end of the outlet tubing from the backpressure regulator through the channel clips along the front right-hand side of the system 2-24 System Setup...
Page 57: Plumbing A Pda Detector
Close the front panel door of the TUV detector. Plumbing a PDA detector If your system includes a PDA detector, see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide for information on plumbing it. Plumbing an ELS detector...
Page 58: Plumbing The Binary Solvent Manager And Sample Manager
Plumbing the binary solvent manager and sample manager To prevent contamination, wear clean, chemical-resistant, Caution: powder-free gloves when plumbing the binary solvent manager and sample manager. To plumb the binary solvent manager and sample manager Route the solvent inlet tubing attached to the in-line degasser and seal wash pump through the channel clips of the sample manager.
Page 59 Remove the 2 protective screws from the corresponding wash ports on the in-line degasser of the binary solvent manager. From the sample manager, route the weak and strong needle wash lines, indicated by the orange and white labels, to their corresponding ports on the in-line degasser in the binary solvent manager.
Page 60 route them through the pass-through of the upper binary solvent manager drip tray. Needle clean system waste port Process waste port Tygon tubing Pharmed tubing TP02590 Connect the PharMed tubing to the front barbed fitting and the Tygon tubing to the front boss fitting, both of which are on the lower binary solvent manager drip tray.
Page 61 10. Remove the protective O-ring from the stainless steel pump outlet tubing. 11. Seat the end of the tubing with the shorter compression screw into the binary solvent manager mixer outlet, and tighten the compression fitting using the 1/4-inch and 5/8-inch open-end wrenches. When using new fittings, tighten 3/4-turn beyond finger-tight.
Page 62 13. Remove the O-ring, and seat the end of the tubing with the longer compression screw into port 5 on the injection valve. Tighten the compression fitting using the 1/4-inch open-end wrench. Injection valve connections Injector outlet/column stabilizer tubing into port 6 Pump outlet tubing into port 5...
Page 63 16. Hold the back of the drain cup, and then slide a waste line over the barbed drain fitting and route it to a suitable waste container. Drip tray Drain cup Barbed drain fitting Waste line Degasser vent line To avoid releasing solvent vapors into the room, route Warning: the degasser vent line to a fume hood or other suitable exhaust system, or to a suitable waste container, ensuring the tubing's...
Page 64 17. Route the degasser vent line to a suitable waste container. Correct positioning of waste and degasser vent lines Waste line Waste line Degasser vent line Degasser vent line Correct Incorrect - will likely TP02709 cause fluid backup 18. Attach a waste line to the barbed fitting on the filter drainage assembly (on the lower front side of the sample organizer), and route it to a suitable waste container.
Page 65: Plumbing The Column Manager
To avoid spills, empty the waste container at regular Warning: intervals. 19. Route a waste line from the barbed fitting on the rear of the solvent tray to a suitable waste container. 20. Close the sample organizer door, sample manager door, sample manager fluidics tray, and the binary solvent manager door.
Page 66: Plumbing The 30-Cm Column Heater/Cooler
Use the gold fittings to connect the stainless steel inlet line to port 6 on the injector valve. Stainless steel inlet line into port 6 Plumbing the 30-cm column heater/cooler If your system includes a 30-cm column heater/cooler, follow the procedure below to plumb it.
Page 67 To plumb the 30-cm column heater/cooler Connect the stainless steel inlet tube from the column to the inject valve port 6. Inject valve port 6 Tubing clips Inlet tube Connect the red PEEK tubing from the outlet of the column to the detector.
Page 68: Routing System Drainage Through The Flexcart
Slide the drip tray onto the bottom of the column compartment, inserting the L-tabs on each side of the drip tray into the slots in the column compartment floor. Column compartment floor Drip tray Slide L-tabs into slots Route the tubing to a suitable waste container. Routing system drainage through the FlexCart To route system drainage through the FlexCart Place a suitable waste container in the bottom right-hand side of the...
Page 69: Installing The Column Stabilizer Assembly In An Ht Column Heater
Route the waste and degasser vent lines (located at the front, bottom right-hand side of the binary solvent manager) down through the cart’s top access hole to the waste container. Degasser vent line Waste line Access hole Route the waste line (located at the front, bottom right-hand side of the sample organizer) down through the cart’s top access hole to the waste container.
Page 70 To install the column stabilizer assembly Open the HT column heater door, lift up and rotate the metal latch clockwise, and tilt the column heater tray 45 degrees downward. Column heater tray Metal latch Remove the clear, protective backing from the thermal gasket, and with the thermal gasket oriented vertically, center it between the two threaded holes.
Page 71 With the column stabilizer assembly’s inlet tubing on the right-hand side, firmly press the assembly into the thermal gasket. Ensure that the thermal gasket uniformly envelops the sides of the column stabilizer and does not bunch beneath it. Thermal gasket visible along both sides of stabilizer Column...
Page 72: Installing The Column Stabilizer Assembly In A Column Heater/Cooler
Carefully route the column stabilizer inlet tubing through the groove on the right-hand side of the tray. To avoid stripping the threads, do not overtighten screws. Caution: Insert 2 Phillips screws into the threaded holes, and tighten them gently using the Phillips screwdriver. Remove the O-ring retainer from the inlet end of the column stabilizer tube, and then connect the inlet tube to injector port 6 of the sample manager.
Page 73 To install the column stabilizer assembly Swing the column retainer rod out to the right. Column retainer rod Remove the clear, protective backing from the thermal gasket, and then center the thermal gasket vertically between the two threaded holes. Press the thermal gasket into the column compartment sleeve. Ensure that the gasket is centered and formed into the bottom of the sleeve with equal lengths on both sides.
Page 74 Column stabilizer assembly (two styles) For 150-mm column For 50-mm or 100-mm column Reusable fittings Inlet tubing to injector TP02874 Carefully route the column stabilizer inlet tubing through the groove on the right-hand side of the tray. Remove the O-ring retainer from the inlet end of the column stabilizer tube, and then connect the inlet tube to injector port 6 of the sample manager.
Page 75: Installing The Column In-Line Filter In A Ht Column Heater
Installing the column in-line filter in a HT column heater You can install a 0.2-μm in-line filter to protect the analytical column against contamination. Required materials • Column in-line filter • Reusable fitting For instruction on how to replace the filter in the HT column heater’s in-line filter unit, see “Replacing the frit in the HT column heater’s in-line filter unit”...
Page 76: Installing The Column In-Line Filter In A Column Manager Or Column Heater/Cooler
Remove the column in-line filter unit from its packaging, and then push the outlet end of the column stabilizer tubing assembly into the in-line filter inlet until it stops. Flow In-line filter unit From column stabilizer Column tubing assembly Inlet TP02873 Reusable fitting Holding the reusable fitting in place, rotate the in-line filter unit onto...
Page 77 To install the column in-line filter Assemble the reusable fitting, collet, and ferrule (in that order) on the outlet end of the column stabilizer tubing assembly, leaving the ferrule 3 mm from the end. Before connecting, make sure that the ferrule is approximately Tip: 3 mm from the end of the outlet tube and that the collet is touching the back of the ferrule.
Page 78: Installing The Column In An Ht Column Heater
Insert the spiked end of the collet separator into one of the holes in reusable fitting, and push the reusable fitting into the column compartment sleeve. To avoid damaging the tubing, use only your fingers to Caution: tighten the tubing fittings. Holding the reusable fitting in place with the spiked end of the collet separator, rotate the in-line filter onto the fitting.
Page 79 To connect the column in-line filter unit and the column inlet Remove the O-ring from the outlet end of the column in-line filter unit. Remove the plug from the column inlet. Push the outlet end of the column in-line filter unit into the column inlet until it stops.
Page 80 .0025-inch ID inlet tubing (included with the flow cell or the stabilizer kit) to the column outlet. Outlet TP02952 Detector inlet tubing Inlet (PEEK) from column Snap the column support clips onto the body of the column (one between the reusable fitting and column). Rotate the clips so that the openings face forward.
Page 81: Installing The Column In A Column Manager Or Column Heater/Cooler
To avoid electric shock or possible damage to the Warning: components, power-off the sample manager before you connect or disconnect the external communication cable (between the sample manager and the HT column heater). To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to an instrument.
Page 82 Push the outlet end of the column in-line filter unit into the column inlet until it stops. To avoid damaging the tubing, use only your fingers to Caution: tighten the fittings. Tighten the fitting until it is snug, and then add 1/4-turn. To install the PEEK tubing in the column outlet To avoid bandspreading, ensure that the tubing is fully Caution:...
Page 83 Using a 5/16-inch wrench to hold the column steady, use the high-temperature outlet fitting wrench to give the outlet fitting a 1/4-turn. Outlet fitting wrench 5/16-inch Column wrench Push the PEEK tubing into the left-hand side of the column tray. To connect the PEEK tubing to the detector Remove the protective cover from the PEEK tubing from the flow cell inlet in the detector.
Page 84 Snap the column support clips onto the body of the column (one between the reusable fitting and column). Rotate the clips so that the openings face forward. Place the column in the column compartment sleeve. Swing the column retainer rod to the left to fasten it. Attach the eCord fob to the receptacle on the right-hand side of the column manager or column heater/cooler.
Page 85: Installing The Column In A System With A Mass Spectrometer
Installing the column in a system with a mass spectrometer If your system includes a mass spectrometer positioned to the right-hand side of the system stack, swing the column heater door to the “away” position (that is, swing it rightward). In this way, you connect to the mass spectrometer’s inlet without increasing the tubing length.
Page 86 Column heater door swung to right the right-hand side Column heater door Rear cover Pull off the rear cover, and then tilt the column tray downward. Column heater door with rear cover removed and column tray open Column heater door Column tray open Snap the plastic column clips onto the body of the column.
Page 87: Connecting To The Solvent Supply
Choose solvent reservoirs that snugly fit the reservoir caps supplied in the startup kit. Waters recommends 1-L reservoirs. Plumbing the system 2-55...
Page 88 Wear clean, chemical-resistant, powder-free gloves Caution: when handling the solvent filter. Skin oils can contaminate the solvent filter. Remove the solvent filter from the short piece of PTFE tubing. Solvent filter PTFE tubing Solvent tubing TP02484 Insert the solvent tubing through the solvent reservoir cap. Re-install the solvent filter on the short piece of PTFE tubing.
Page 89: Making Ethernet And Signal Connections
The HT column heater, column manager, or column heater/cooler incorporates an internal Ethernet switch, that accommodates the PC (workstation) and up to six ACQUITY UPLC modules. Connect the shielded Ethernet cables from each module to the electronic connections on the rear panel of the HT column heater, column manager, or column heater/cooler.
Page 90: 30-Cm Column Heater/Cooler Connections
Connect the external communication cable to the High Density (HD) port on the rear of the HT column heater. Connect the other end of the external communication cable to the QSPI port on the rear of the sample manager. 30-cm column heater/cooler connections To make 30-cm column heater/cooler connections To avoid damaging electrical parts, never disconnect an Caution:...
Page 91: Binary Solvent Manager I/O Signal Connectors
Ethernet cable to the network switch, and the other end to the Ethernet card on the workstation. Waters Ethernet Instrument Getting Started Guide. See also: Plug the 8-port network switch box into a suitable wall outlet.
Page 92 Binary solvent manager I/O signal connectors Connector I Connector II Auxiliary 1 In Start Gradient In − Auxiliary 1 In − Start Gradient In Auxiliary 2 In Stop Flow − Auxiliary 2 In − Stop Flow Ground Ground Run Stopped Out Switch 2 Out Run Stopped Out Switch 2 Out...
Page 93: Sample Manager I/O Signal Connectors
Binary solvent manager analog-out/event-in connections (Continued) Signal connection Description Switch 2 Out Used to send time-based contact closure signals to external devices. Switch 3 Out Used to send time-based contact closure signals to external devices. 0−2V Analog 2 Out Outputs the analog signal to a device such as an integrator or strip-chart recorder.
Page 94 Sample manager I/O signal connectors Connector I Connector II Inject Start Switch 1 Out − Inject Start Switch 1 Out Auxiliary In Switch 2 Out − Auxiliary In Switch 2 Out Ground Switch 3 Out Inject Hold In Switch 3 Out −...
Page 95: Tuv Detector I/O Signal Connectors
Sample manager analog-out/event-in connections (Continued) Signal connections Description Run Stopped Out Indicates (with a contact closure) that the sample manager ceased operation because of an error condition or an operator request. TUV detector I/O signal connectors The rear panel of the TUV detector includes two removable connectors that hold the screw terminals for I/O signal cables.
Page 96: Pda Detector Signal Connectors
PDA detector signal connectors If your system includes a PDA detector, see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide for information on signal connectors. ELS detector signal connectors If your system includes an ELS detector, see the ACQUITY UPLC Evaporative Light Scattering Detector Getting Started Guide for information on signal connectors.
Page 97: Connecting To The Electricity Source
To make signal connections, attach the positive and negative leads of the signal cable to the connector. Connector Signal cable Connecting to the electricity source Each system instrument requires a separate, grounded power source. The ground connection in all power outlets must be common and physically close to the system.
Page 98 FlexCart power connections To circuit A AC line Network switch Detector Sample manager Binary solvent FlexCart manager power strips Sample organizer LCD/monitor To circuit B AC line 2-66 System Setup...
Page 99: Calibrating The Xyz Mechanism Using The Teach Block
Calibrating the XYZ mechanism using the teach block Before performing any chromatography, you must verify that the XYZ mechanism positions the needle properly in the sample chamber. If your system has a sample organizer, you must install the Requirement: organizer before calibrating the XYZ mechanism. To avoid puncture wounds, keep hands or loose clothing Warning: clear of the needle assembly mechanism while it is moving.
Page 100 Insert the teach block into the locating holes. Teach block in position In the ACQUITY UPLC Console, select Sample Manager from the system tree. Click Troubleshoot > Calibrate XY and Zp axes. Calibrate XY and Zp Axes dialog box 2-68...
Page 101 In the Calibrate XY and Zp Axes dialog box, click Start to begin. A warning appears, prompting you to keep clear of the sample Result: chamber. Confirm that all sample plates and trays are removed, and ensure the teach block is installed. In the warning dialog box, click OK.
Page 102 13. To laterally position the puncture needle, use the +X and −X buttons to move it to the left-hand or right-hand side, respectively. Press the keyboard arrow keys instead of clicking the Alternative: on-screen buttons to adjust the needle position. Doing so lets you adjust the position without taking your eyes off the needle.
Page 103 20. Click Maintain > Calibrate needle Z axis. Calibrate Needle Z Axis dialog box 21. In the Calibrate Needle Z Axis dialog box, click Start, and then click OK in the confirmation window. 22. Use the +Z button to drive the sample needle down to within 1 mm of the tray surface.
Page 104 To easily and efficiently achieve the correct needle setting, slide a Tip: business card under the needle. Then lower the needle until it touches the card lightly but does not restrict the card’s free movement. Sample needle almost touching surface of sample tray holder or just touching the business card 24.
Page 105: Preparing System Hardware
You must power-on the HT column heater, column manager, or column heater/cooler before any of the other instruments because they contain the internal Ethernet switch that allows the ACQUITY UPLC system workstation to communicate with all the instruments. Powering-on the system...
Page 106 If your system contains an HT column heater, it is automatically Tip: powered-on when you power on the sample manager. To power-on a column manager or column heater/cooler, press the power button on the top, left-hand side of it’s front panel. Each system instrument beeps 3 times and runs a series of startup tests.
Page 107: Monitoring Startup Tests
Start the Empower or MassLynx operating software. You can monitor the ACQUITY UPLC Console for messages and LED indications. Monitoring startup tests These startup tests run when you power-on the ACQUITY UPLC system workstation: • CPU board •...
Page 108: Power Led
Power LED The power LED, on the left-hand side of an instrument’s front panel, indicates the power-on or power-off status of the instrument. This LED is green when power is on and unlit when power is off. To provide adequate ventilation, the sample manager and sample Tip: organizer fans are always running, even with the power off.
Page 109 • Detector – Indicates the detector is initializing or calibrating. Flashing red Indicates that an error stopped the instrument. Refer to the ACQUITY UPLC Console for information regarding the error. Steady red Indicates an instrument failure that prevents further operation. Power-off the instrument, and then power-on.
Page 110: Enabling The Leak Sensors
When you power-on the system, the leak sensors default to disabled Rule: unless previously enabled. To enable the leak sensors In the ACQUITY UPLC Console, select Control > Leak Sensors. Leak Sensors dialog box Click to enable or disable all...
Page 111: Preparing The Binary Solvent Manager
Preparing the binary solvent manager For optimal performance of the ACQUITY UPLC system, you must prepare the binary solvent manager for operation. To prepare the binary solvent manager for operation, you must perform a seal wash prime and then prime the binary solvent manager.
Page 112: Required Materials
Before priming the plunger seals, ensure the volume of seal wash is adequate for priming. Controlling Contamination in Ultra Performance LC/MS and See also: HPLC/MS Systems (part number 715001307) on the ACQUITY UPLC System Bookshelf CD. Required materials • 30-mL syringe (startup kit) •...
Page 113: Priming The Binary Solvent Manager
In the ACQUITY UPLC Console, select Binary Solvent Manager from the system tree. Click Control > Prime seal wash, and then click Yes to begin the seal wash priming process. Slowly draw back on the syringe plunger to pull seal wash solvent through the system.
Page 114: Priming A Dry Binary Solvent Manager
To prime a dry binary solvent manager Open the instrument’s front door. Locate the appropriate solvent vent line. In the ACQUITY UPLC Console, select Binary Solvent Manager from the system tree. In the binary solvent manager information window, click Control >...
Page 115 PharMed tubing to the solvent vent line you located in step In the ACQUITY UPLC Console, select Binary Solvent Manager from the system tree. In the binary solvent manager information window, click Control >...
Page 116: Priming A Wetted Binary Solvent Manager
Prime the binary solvent manager for 4 minutes if you will use new solvents that are of the same composition of what is already in the system. To refresh the system In the ACQUITY UPLC Console, click Control > Refresh system (Sys Prep). 3-12 Preparing System Hardware...
Page 117 To start up the system In the ACQUITY UPLC Console, click Control > Start up. In the Prime Solvents tab of the System Startup dialog box, review the settings for the A/B Solvents. In the A/B Solvents area, you can select or clear any or all of the solvents: A1, A2, B1, or B2.
Page 118 If you changed the sample loop, in the Optional: Characterize Volume tab, select Characterize needle and loop volumes. Click Start. The lamp in the optical detector ignites, the ACQUITY UPLC Result: system sets the column sample temperatures, and all priming starts. If...
Page 119: Preparing The Sample Manager
25% methanol or acetonitrile and a strong wash solvent of 50% to 100% methanol or acetonitrile. High sample concentrations can require other weak wash solvents. If your separation permits, Waters recommends adding a small amount of an organic solvent (~10%) to prevent microbial growth.
Page 120 Appendix C for further information about solvents. To avoid damage to the solenoid valve seats and seals in the Caution: solvent path, do not use a nonvolatile buffer as the weak wash or strong wash solvent. For best performance, the weak wash solvent must be similar or identical Tip: to your isocratic or initial gradient solvent conditions, excluding buffers.
Page 121: Priming The Sample Manager
The sample manager must be primed before you attempt to Requirement: characterize the seal. To prime the sample manager In the ACQUITY UPLC Console, select Sample Manager from the system tree. Preparing the sample manager 3-17...
Page 122 However, do not select this option routinely. Priming all components at the same time is good practice. Type the number of primes in the Number of cycles text box. Default: Waters recommends 5 to 7 primes when you are Recommendation: changing solvents. Do not abort the sample manager priming sequence.
Page 123: Washing The Sample Manager Needle
Tip: the system, you can omit this procedure. Observing wash solvent recommendations Waters recommends that you observe these guidelines for washing the needle: • To ensure that the strong wash solvent is completely removed, the system washes the needle with 500 μL of weak wash solvent after you use strong wash solvent.
Page 124 Needle recommendations Waters offers needles made from several types of materials to accommodate compounds that may have specific handling requirements or affinities. Users must choose the needle material best suited to their application. For example, the stainless steel needle is recommended when working with samples that are known to be attracted to hydrophobic polymers.
Page 125: Characterizing The Needle Seal
In the Weak Wash box, specify the volume for the weak wash solvent. 1.0 through 99,999 μL Range: 200.0 μL without strong wash or 500 μL with strong wash Default: 200 through 500 μL or three times the strong wash Recommendation: volume.
Page 126: Characterizing The Needle And Sample Loop Volumes
To characterize the needle seal In the ACQUITY UPLC Console, select Sample Manager from the system tree. In the sample manager information window, click Maintain > Characterize > Needle seal. In the Characterize Needle Seal dialog box, click Start. The calibrate seal operation begins, and the sample manager status displays “Calibrating seal”.
Page 127: Using The Extended Puncture Needle
If you are running under MassLynx software control, click Inlet Method > Inlet > Autosampler in the Inlet Editor window. To characterize the needle and sample loop volumes In the ACQUITY UPLC Console, select Sample Manager from the system tree. Click Maintain > Characterize > Needle and loop volumes.
Page 128: Loading Sample Plates In The Sample Manager
Loading sample plates in the sample manager The ACQUITY UPLC sample manager holds up to two ANSI/SBS plates that you load through the front door. The left-hand plate is referred to as position 1, the right-hand plate as position 2.
Page 129: Selecting The Optimum Sample Injection Mode
Load the plate onto the tray so that well position A,1 is at the rear, right-hand corner and the forward edge of the plate is behind the spring inside the front of the carrier. A represents the row number, 1 represents the vial position. Tip: Slide the tray into the sample manager until it clicks into place.
Page 130 The first injection of a sample set and injection sets with different methods do not utilize load ahead mode. The minimum cycle time is the lesser of either the two run times or sample preparation and wash. The following figures show a standard ACQUITY UPLC system cycle time and load-ahead cycle time. 3-26...
Page 131 Standard ACQUITY UPLC system cycle time definition Standard injection mode and first injection in load ahead mode Total ACQUITY UPLC system cycle time System setup Chromatographic run time SM sample preparation and positioning SM wash time Sample injection Start ACQUITY UPLC system load-ahead cycle time definition...
Page 132: Installing The Optional Sample Manager Shade
If your samples are light-sensitive, and you have the optional sample manager foam shade, be sure to install it over the sample compartment window. For the sample manager foam shade part number, see the Waters Quality Parts Locator on the Waters Web site’s Services/Support page.
Page 133 Insert the foam shade over the outside of the sample compartment window. Sample compartment window Foam shade Press the foam shade into place over the window. Foam shade Preparing the sample manager 3-29...
Page 134: Preparing The Column Manager
Exception: left-hand tray in the sample manager is designated position 1 and the right-hand tray is position 2. In the ACQUITY UPLC Console, select Sample Manager from the system tree. In the sample manager information window, click Configure > Sample Organizer.
Page 135: Loading Sample Plates
Shelf label TP02504 To prevent spillage, use Waters-approved cap mats, Caution: sealing caps, or heat seal film on the samples. Consult the current ACQUITY UPLC system release notes for a list of approved sample covers. Preparing the sample organizer 3-31...
Page 136 Ensure that the plate does not extend beyond the plate stop at the rear of the shelf. To ensure the transfer shuttle moves freely and without Caution: damaging the sample organizer, you must be able to slide a shelf/plate/vial combination in or out without interfering with the shelves or plates directly above and below it.
Page 137 Caution: sealing caps, or heat seal film on the samples. Consult the current ACQUITY UPLC system release notes for a list of approved sample covers. Load a plate onto a shelf so that position A,1 is at the rear, right-hand corner and the forward edge of the plate is behind the stop at the front, left-hand corner.
Page 138 Caution: sealing caps, or heat seal film on the samples. Consult the current ACQUITY UPLC system release notes for a list of approved sample covers. Load a plate onto a shelf so that position A,1 is at the rear, right-hand corner and the forward edge of the plate is behind the stop, at the front, left-hand corner.
Page 139: Displaying Sample Plate Information
In the ACQUITY UPLC Console, a thin, gray bar appears for each Tip: empty shelf. After the plate or shelf is configured using Empower or MassLynx software, plate identification appears in a thicker bar. Configure the plates and shelves in the Empower or MassLynx data application.
Page 140: Starting The Tuv Detector
If your system includes a TUV detector, prepare it for operation by following the procedures in this section. If your system includes a PDA detector, see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide for information on preparing it.
Page 141 3 minutes. When the lamp LED shows steady green, start the Empower or MassLynx software. You can monitor the ACQUITY UPLC Console for messages and LED indications. For best results, allow at least 30 minutes to equilibrate the detector and stabilize the baseline.
Page 142 TUV detector information window Recording sample and reference energies After you install the detector or perform maintenance tasks, like changing the lamp or flow cell, complete the procedures in this section to verify that the detector optics and electronics work properly. To record sample and reference energies Ensure that the detector is connected to the workstation.
Page 143: Conditioning The Column
Ensure the detector cell is filled with solvent and free of air bubbles. When both LEDs show steady green, initialization is complete. Start the Empower or MassLynx software. Launch the ACQUITY UPLC Console from the sample manager control panel. For additional information, see “Starting the ACQUITY UPLC Console...
Page 144: Shutting Down The System
If MassLynx software controls the system, proceed as follows: Open the Sample Set window, and select an inlet method that includes the chromatographic conditions you want to use. Run the sample set line. The system runs the separation method. For more information about column conditioning, consult the See also: Empower or MassLynx online Help.
Page 145: Shutting Down For More Than 24 Hours
Flush the column and flow cell with 100% pure organic solvent. Waters ACQUITY UPLC BEH Column Care and Use See also: Instructions or ACQUITY UPLC HSS Column Care and Use Instructions. Risk of electric shock. The power switch on each system Warning: instrument controls the basic operational state of that instrument.
Page 146: Running Hplc Methods On An Acquity Uplc System
HPLC methods to UPLC methods. System considerations Traditional HPLC methods run on an ACQUITY UPLC system can differ in retention time—and in some cases peak order—from the same methods run on traditional HPLC equipment. The differences are primarily due to lower system volume and reduced bandspreading in the ACQUITY UPLC system, which operates with a maximum injection volume of 50 μL and a maximum...
Page 147 Reusable fittings Inlet tubing to injector TP02874 • Do not use the ACQUITY UPLC column in-line filter unit with an HPLC column. • Set the high pressure limit of the binary solvent manager to the maximum pressure recommended for the HPLC column.
Page 148: Choosing Fittings
To reduce variability from temperature fluctuations, use a Recommendation: column oven with a thermostat. For columns longer than 150 mm, use the ACQUITY UPLC system 30-cm column heater/cooler or another appropriate external oven. If you observe a discrepancy between the selectivity and resolution of the...
Page 149 Column outlet If your system has a Waters-supplied, wrench-flat, column outlet fitting that has been used for a UPLC column connection, replace the swage fitting with a reusable fitting of your choice, or a swage fitting recommended by the HPLC column manufacturer.
Page 150 3-46 Preparing System Hardware...
Page 151: Configuring System Software
Configuring System Software Contents Topic Page Configuring Empower software Starting the ACQUITY UPLC Console from Empower software 4-12 Configuring MassLynx software 4-13 Starting the ACQUITY UPLC Console from MassLynx software 4-14 Configuring Empower software Perform these tasks to configure Empower software: •...
Page 152: Selecting System Instruments
Selecting system instruments To select system instruments In the Empower Pro window, click Configure System. In the Configuration Manager window, click Acquisition Servers, right-click the node name, and then select Properties. If you are using Empower 1154 software to control the system, the Tip: node name is referred to as the acquisition server name.
Page 153 System Selection dialog box In the System Selection dialog box, drag the name of the instrument(s) you want to include in the new system from the Available Instruments pane to the New System Instruments pane. Click Next. When the Access Control dialog box appears, click Next. In the Name Selection dialog box, specify a name for your system.
Page 154: About The Binary Solvent Manager Control Panel
Control panels About the binary solvent manager control panel If Empower software controls the system, the binary solvent manager’s control panel appears at the bottom of the Run Samples window. If MassLynx software controls the system, the binary solvent manager's control panel appears on the Additional Status tab of the Inlet Editor window.
Page 155 System Pressure Displays the binary solvent manager pressure, in kPa, bar, or psi. Pressure units can be customized through the ACQUITY UPLC Console. Total Flow Rate Displays the total flow rate of the binary solvent manager. Total flow rate values range from 0.000 to 2.000 mL/min under normal...
Page 156: About The Sample Manager Control Panel
Reset BSM Resets the binary solvent manager after an error condition. Help Displays the ACQUITY UPLC Console online Help. About the sample manager control panel If Empower software controls the system, the sample manager’s control panel appears at the bottom of the Run Samples window. If Masslynx software controls the system, the sample manager's control panel appears on the Additional Status tab of the Inlet Editor window.
Page 157 Sample manager control panel Run LED Status Current HT column Current sample heater temperature compartment temperature Display ACQUITY UPLC Console Sample compartment HT column heater set point set point The sample manager control panel displays current sample compartment and HT column heater temperatures and set points. You can edit these values when the system is idle by clicking on the underlined value.
Page 158 Displays the current column heater set point to 0.1 °C resolution. When active temperature control is disabled, this field displays “Off”. Displays the ACQUITY UPLC (Display Console) Console. You can access these additional functions by right-clicking anywhere in the sample manager control panel:...
Page 159: About The Tuv Detector Control Panel
You cannot edit detector parameters while the system is running samples. If your system includes a PDA detector, see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide for information on the control panel. If your system includes an ELS detector, see the ACQUITY UPLC Evaporative Light Scattering Detector Getting Started Guide for information on the control panel.
Page 160 Additional functions in the detector control panel Control panel function Description Autozero Resets the absorbance value to 0 Reset TUV Resets the detector, when present, after an error condition Help Displays the ACQUITY UPLC Console online Help 4-10 Configuring System Software...
Page 161: About The Column Manager Control Panel
About the column manager control panel If Empower software controls the system, the column manager’s control panel appears at the bottom of the Run Samples window. If Masslynx software controls the system, the column manager's control panel appears on the Additional Status tab of the Inlet Editor window.
Page 162: Starting The Acquity Uplc Console From Empower Software
Displays the ACQUITY UPLC Console online Help Starting the ACQUITY UPLC Console from Empower software To start the ACQUITY UPLC Console from Empower software In the Run samples window, click Display console in the Sample Manager control panel. ACQUITY UPLC Console window...
Page 163: Configuring Masslynx Software
Tools menu of the Inlet Method window. In the Inlet Configuration window, click Configure and then click Next. In the Select Pump dialog box, select Waters ACQUITY as the pumping device, and then click Next. Select Waters ACQUITY as the autosampler, and then click Next.
Page 164: Starting The Acquity Uplc Console From Masslynx Software
13. Click Finish. The Inlet Method window appears. Starting the ACQUITY UPLC Console from MassLynx software To start the ACQUITY UPLC Console from MassLynx software In the MassLynx window, click Inlet Method. In the Inlet Method window, click the ACQUITY Additional Status tab.
Page 165: Verifying System Operation
Performing the gradient performance test 5-10 Restrictions: ® • This gradient performance test is not applicable to ACQUITY UPLC systems that have a mass spectrometer. • There is no gradient performance test for ACQUITY UPLC systems that have an ELS detector.
Page 166: Preparing The System
• All MS-grade solvents used with the ACQUITY UPLC system must be properly filtered prior to their use. Waters recommends that the solvent be filtered through an appropriate .22-μm or smaller membrane filter using a solvent clarification kit (47-mm all-glass filter holder with 1-L flask) immediately before use.
Page 167 To prepare the system for verification To avoid chemical exposure risk, always observe Good Warning: Laboratory Practices when you use this equipment and when you work with solvents and test solutions. Know the chemical and physical properties of the solvents and test solutions you use. See the Material Safety Data Sheet for each solvent and test solution in use.
Page 168 “Priming the binary solvent manager” on page 3-9. Install the ACQUITY UPLC column in the column heater, close the column tray, and replace the column heater’s front cover. If you need more information about installing the column, see “Installing the column in an HT column heater”...
Page 169: Creating The Test Methods
Creating the test methods The gradient performance test method parameters are the same whether Empower or MassLynx software controls the system. Follow the steps below to create the methods, setting the parameter values to match those pictured in the screen representations. Click on the tabs to display online Help.
Page 170 The binary solvent manager parameters are the same regardless of Tip: whether your system includes the TUV or PDA detector. Set instrument method parameters for the sample manager as shown in the following screen representation. Sample manager instrument parameters The sample manager parameters are the same regardless of Tip: whether your system includes the TUV or PDA detector.
Page 171 Sample manager instrument advanced settings The sample manager parameters are the same regardless of Tip: whether your system includes the TUV or PDA detector. Creating the test methods...
Page 172 For more information on detector parameters, consult the ACQUITY UPLC Console online Help, Empower online Help, or MassLynx online Help. • If your system includes a PDA detector, see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide for information on verifying it. •...
Page 173: Creating The Sample Set Method
• If your system includes an FLR detector, see the ACQUITY UPLC Fluorescence Detector Getting Started Guide for information on verifying it. • If your system includes an SQ detector, see the SQ Detector Operator’s Guide for information on verifying it.
Page 174: Performing The Gradient Performance Test
Performing the gradient performance test If your ACQUITY UPLC system uses only a mass spectrometer, Restriction: the gradient performance test is not applicable. When the system is prepared and the test methods are created, you are ready to perform the gradient performance test. The steps for running the test vary slightly, depending on whether your system uses Empower or MassLynx software, but the desired results are the same.
Page 175 Review the gradient performance report. The gradient performance test result is “passing” when these conditions are realized: • The peaks are symmetrical, integrated, and identified correctly. (Compare the chromatogram on the report to the sample chromatogram, below.) • The peak retention times show a standard deviation of ≤ 2.0 s. (Consult the table you completed.) Sample gradient performance test chromatogram 0.34...
Page 176 5-12 Verifying System Operation...
Page 177: Maintaining The System
6-225 Maintaining the 30-cm column heater/cooler 6-228 If your system includes a PDA detector, see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide for information on maintaining it. If your system includes an ELS detector, see the ACQUITY UPLC Evaporative Light Scattering Detector Getting Started Guide for information on maintaining it.
Page 178: Maintenance Schedule
Maintenance schedule Waters recommends that you perform the following routine maintenance on ® the ACQUITY UPLC system to ensure reliable operation and accurate results. When using the system throughout the day (and on nights and weekends), or when using aggressive solvents such as buffers, perform these maintenance tasks more frequently.
Page 179 Recommended routine maintenance schedule (Continued) Maintenance procedure Frequency For information... Replace the accumulator During scheduled page 6-46 check valve on the binary preventive solvent manager maintenance or as needed Replace solvent filters During scheduled page 6-49 preventive maintenance or as needed Clean the air filters in the As needed...
Page 180 Recommended routine maintenance schedule (Continued) Maintenance procedure Frequency For information... Characterize the needle and Whenever the needle page 6-140 sample loop volumes on the or sample loop is sample manager replaced Replace the puncture needle As needed page 6-142 holder on the sample manager Replace the sample loop on During scheduled page 6-145...
Page 181: Spare Parts
6-161, paper dampened with water page 6-225, page 6-228, and page 6-230 Spare parts Replace only the component parts mentioned in this document. See the Waters Quality Parts Locator on the Waters Web site’s Services & Support page. Spare parts...
Page 182: Contacting Waters Technical Service
Waters Technical Service (800 252-4752). Otherwise, phone the Waters corporate headquarters in Milford, Massachusetts (USA), or contact your local Waters subsidiary. Waters’ site includes phone numbers and e-mail addresses for Waters locations worldwide. Visit www.waters.com, and click Waters Division > Regional/Global Contacts.
Page 183: Maintenance Considerations
To view instrument information In the ACQUITY UPLC Console, select an instrument from the system tree. Click Configure > View module information. The Module Information dialog box displays this information: • Serial Number • Firmware Version • Firmware Checksum •...
Page 184: Proper Operating Procedures
For information on setting maintenance warnings, consult the ACQUITY UPLC Console online Help. Maintenance warnings can be set for these conditions: •...
Page 185: Maintaining The Binary Solvent Manager
Perform the procedures in this section when you discover a problem with a binary solvent manager component or during preventive maintenance. For information about isolating problems in the binary solvent manager, consult the ACQUITY UPLC Console online Help. To prevent injury, always observe Good Laboratory Warning: Practices when you handle solvents, change tubing, or operate the binary solvent manager.
Page 186: Resolving Binary Solvent Manager Leak Sensor Errors
Binary solvent manager major components Pressure transducer Primary Accumulator Primary pump A pump A cable connectors pump B Accumulator pump B Valve Valve actuator B actuator A Degasser Drip tray Degasser vent line Mixer Seal wash Vent valve Seal wash drip tray tube Drain tube Solvent...
Page 187: Required Materials
Nonabrasive, lint-free wipes To resolve a binary solvent manager leak sensor error View the Leak Sensors dialog box in the ACQUITY UPLC Console to confirm that the binary solvent manager leak sensor detected a leak. If a leak is detected, a “Leak Detected” error message appears.
Page 188 Turn the vent tube retainer clockwise, and then lift the A-vent and B-vent tubes out of the drip tray by pulling up on them and moving them to the left-hand side of the leak sensor. Turn vent tube retainer clockwise A-vent and B-vent tubes Leak sensor TP02884...
Page 189 Use a nonabrasive, lint-free wipe to dry the leak sensor prism. Prism Lint-free wipe TP02891 Roll up a nonabrasive, lint-free wipe, and use it to absorb the liquid from the leak sensor reservoir and its surrounding area. Leak sensor reservoir Rolled up lint-free wipe Maintaining the binary solvent manager...
Page 190 With a cotton swab, absorb any remaining liquid from the corners of the leak sensor reservoir and its surrounding area. Leak sensor reservoir Cotton swab 10. Align the leak sensor’s T-bar with the slot in the side of the leak sensor reservoir, and slide the leak sensor into place.
Page 191: Replacing The Binary Solvent Manager's Leak Sensor
13. If you detached the connector from the front of the instrument, reattach 14. Power-on the binary solvent manager. 15. In the ACQUITY UPLC Console, select Binary Solvent Manager from the system tree. 16. In the binary solvent manager information window, click Control >...
Page 192 Required materials • Clean, chemical-resistant, powder-free gloves • Leak sensor To replace the binary solvent manager’s leak sensor To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to an instrument. To completely interrupt power to an instrument, set the power switch to Off, and then unplug the power cord from the AC outlet.
Page 193 Turn the vent tube retainer clockwise, and then lift the A-vent and B-vent tubes out of the drip tray by pulling upward on them and moving them to the left-hand side of the leak sensor. Turn vent tube retainer clockwise A-vent and B-vent tubes Leak sensor TP02884...
Page 194 Align the leak sensor’s T-bar with the slot in the side of the leak sensor reservoir, and slide the leak sensor into place. TP02892 TP02889 T-bar Slot in leak sensor reservoir Leak sensor installed in reservoir TP02888 Reinsert the A-vent and B-vent tubes into the drip tray. Turn the vent tube retainer, which holds the A-vent and B-vent tubing in place, counterclockwise.
Page 195: Replacing The Mixer
10. Connect the leak sensor connector to the front of the instrument. 11. Power-on the binary solvent manager. 12. In the ACQUITY UPLC Console, select Binary Solvent Manager from the system tree. 13. In the binary solvent manager information window, click Control >...
Page 196 Using the 5/8-inch open-end wrench to hold the mixer in place, disconnect the outlet compression fitting with the 1/4-inch open-end wrench. Mixer Outlet compression fitting 5/8-inch open-end wrench 1/4-inch open-end wrench Again using the 5/8-inch open-end wrench to hold the mixer, disconnect the inlet compression fittings with the 1/4-inch wrench.
Page 197: Replacing The I Valve Actuator
Replacing the i Valve actuator To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the binary solvent manager. Know the physical and chemical properties of the solvents you use. See the Material Safety Data Sheets for the solvents in use. Caution: •...
Page 198 To avoid solvent spills, move the solvent bottles to a Warning: location below the binary solvent manager. Move the solvent bottles to a location below the binary solvent manager. To avoid damage to the connector or cable, grasp the Caution: Valve connector by the knurled diameter.
Page 199 Loosen the cap nut on the in-line filter so that it is all the way off the ferrule holder fitting threads. Cap nut TP02958 Ferrule holder fitting Cap nut Maintaining the binary solvent manager 6-23...
Page 200 Use the 5/16-inch open-end wrench to loosen the shell nut and then fully unscrew it. Shell nut 5/16-inch wrench Caution: • When you remove the valve assembly, ensure the PEEK washer, which is normally on the top face of the i Valve cartridge, does not remain in the head.
Page 201 Remove the i Valve actuator from the bottom of the primary pump head. Bottom of primary pump head Valve actuator TP02961 Remove the ferrule holder fitting from the old i Valve actuator. Ferrule holder fitting TP02960 Use the #0 Phillips screwdriver to loosen 1/2-turn the 4 screws that secure the clamping plates.
Page 202 Tips: • Avoid touching the clamping plate tabs when loosening the screws. • The shell nut can be rotated to access all 4 screws. #0 Phillips screwdriver Shell nut Screw (4) Clamping plate Clamping plate tab (2) TP03022 Clamping plate 10.
Page 203 12. Use the #0 Phillips screwdriver to loosen 1/2-turn the 4 screws that secure the clamping plates on the new i Valve actuator. Ensure the plates are loose and in their maximum open position. Tips: • Avoid touching the clamping plate tabs when loosening the screws. •...
Page 204 The clamping plates must be fully engaged in the Requirement: cartridge groove. Clamping Clamping plate tab plate tab Clamping plates TP02767 15. While squeezing the clamping plate tabs, use the #0 Phillips screwdriver to tighten the 4 screws that secure the plates, following the torquing pattern shown in step #0 Phillips screwdriver...
Page 205 16. Follow the pattern shown below when tightening the screws that secure the plates, and repeat it at least 3 times, gradually increasing the torque until the screws are tight. Start here 17. Insert the ferrule holder fitting into the inlet port on the i Valve assembly, and tighten it finger-tight.
Page 206 condition. Use the 5/16-inch open-end wrench to tighten the nut an additional 1/8-turn. Shell nut 5/16-inch wrench 20. Reinsert the in-line filter and tube into the ferrule holder fitting. Ferrule holder fitting In-line filter 6-30 Maintaining the System...
Page 207 21. Place the cap nut over the ferrule holder fitting and finger-tighten the cap nut to the extent possible. Ferrule holder fitting Cap nut 22. Align the red dot on the i Valve connector with the red dot on the receptacle, in the 12 o’clock position, and insert the connector into the receptacle.
Page 208: Replacing The I Valve Cartridge
Replacing the i Valve cartridge To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the binary solvent manager. Know the physical and chemical properties of the solvents you use. See the Material Safety Data Sheets for the solvents in use. Caution: •...
Page 209 Grasp the i Valve connector by the knurled diameter and pull it toward you to disconnect it from the receptacle. Connector receptacle Valve connector Knurled diameter Maintaining the binary solvent manager 6-33...
Page 210 Loosen the cap nut on the in-line filter so that it is all the way off the ferrule holder fitting threads. Cap nut TP02958 Ferrule holder fitting Cap nut 6-34 Maintaining the System...
Page 211 Use the 5/16-inch open-end wrench to loosen the shell nut and then fully unscrew it. Shell nut 5/16-inch wrench Caution: • When you remove the valve assembly, ensure the PEEK washer, which is normally on the top face of the i Valve cartridge, does not remain in the head.
Page 212 Remove the i Valve actuator from the bottom of the primary pump head. Bottom of primary pump head Valve actuator TP02961 To avoid damaging the i Valve actuator, do not back the Caution: screws out all the way. Use the #0 Phillips screwdriver to loosen 1/2-turn the 4 screws that secure the plates.
Page 213 #0 Phillips screwdriver Shell nut Clamping plate tab (2) Clamping plate Screw (4) TP02788 Clamping plate Remove the cartridge from the i Valve actuator when both plates are in the maximum open position. Ensure that the low pressure gasket has been removed with the cartridge.
Page 214 12. With the clamping plates still open, insert the cartridge into the i Valve actuator, grooved end first. Grooved end of cartridge Valve actuator 13. With one hand, squeeze the 2 clamping plate tabs on the i Valve actuator to hold the clamping plates against the cartridge. The clamping plates must be fully engaged in the Requirement: cartridge groove.
Page 215 14. While squeezing the clamping plate tabs, use the #0 Phillips screwdriver to tighten the 4 screws that secure the plates, following the torquing pattern shown in step #0 Phillips screwdriver Screw (4) TP03034 15. Follow the pattern shown below when tightening the screws that secure the plates, and repeat it at least 3 times, gradually increasing the torque until the screws are tight.
Page 216 condition. Use the 5/16-inch open-end wrench to tighten the nut an additional 1/8-turn. Shell nut 5/16-inch wrench 18. Reinsert the in-line filter and tube into the ferrule holder fitting. Ferrule holder fitting In-line filter 6-40 Maintaining the System...
Page 217 19. Place the cap nut over the ferrule holder fitting and finger-tighten the cap nut to the extent possible. Ferrule holder fitting Cap nut 20. Align the red dot on the i Valve connector with the red dot on the receptacle, in the 12 o’clock position, and insert the connector into the receptacle.
Page 218: Replacing The In-Line Filter Cartridge On The I Valve Actuator
Replacing the in-line filter cartridge on the i Valve actuator To prevent injury, always observe Good Laboratory Warning: Practices when you handle solvents, change tubing, or operate the binary solvent manager. Know the physical and chemical properties of the solvents you use. See the Material Safety Data Sheets for the solvents in use.
Page 219 Unscrew the cap nut on the in-line filter assembly. Ferrule holder fitting Cap nut Valve actuator Pull the cap nut off the tube to remove the in-line filter cartridge. Cap nut Tube In-line filter cartridge Maintaining the binary solvent manager 6-43...
Page 220 In-line filter cartridge and cap nut Cap nut In-line filter cartridge Tube Metal locking ring TP03025 Put the cap nut over the end of the tube. See the figure “Exploded view of in-line filter” on page 6-45. Slide the metal locking ring onto the tube. Ensure that the thicker end of the metal locking ring is facing toward the cap nut.
Page 221 Exploded view of in-line filter Ferrule holder fitting Cap nut In-line filter cartridge Metal locking ring Insert the in-line filter cartridge with tubing into the ferrule holder fitting. Ferrule holder fitting In-line filter 10. Screw the cap nut onto the ferrule holder fitting, ensuring that the tube is bottomed out in the in-line filter cartridge.
Page 222: Replacing The Accumulator Check Valve
Replacing the accumulator check valve To prevent injury, always observe Good Laboratory Warning: Practices when you handle solvents, change tubing, or operate the binary solvent manager. Know the physical and chemical properties of the solvents you use. See the Material Safety Data Sheets for the solvents in use.
Page 223 Using the 5/16-inch open-end wrench to hold the check valve in place, disconnect the compression fitting from the check valve with the 1/4-inch open-end wrench. 5/16-inch open-end wrench Accumulator check valve Compression fitting 1/4-inch open-end wrench When you remove the valve assembly, ensure the PEEK Caution: washer, which is normally on the top face of the check valve, does not remain in the head.
Page 224 Use the 1/2-inch open-end wrench to loosen the check valve, and then remove the check valve assembly from the head. Head Accumulator check valve TP02822 1/2-inch open-end wrench Unpack the new check valve. Make sure the new PEEK washer is inserted into the new check valve so that its chamfered edge faces away from the check valve.
Page 225: Replacing Solvent Filters
Insert the check valve assembly into the head, and use the 1/2-inch wrench to tighten the check valve nut 1/8-turn beyond finger-tight. Using the 5/16-inch open-end wrench to hold the check valve in place. Reattach the compression fitting to the check valve. Use the 1/4-inch wrench to tighten the compression screw 1/4-turn beyond finger-tight for existing stainless steel tubing assembly, or 3/4-turn beyond finger-tight for new stainless steel tubing assembly.
Page 226: Cleaning The Air Filters In The Binary Solvent Manager Door
Insert the new solvent filter into the PTFE tubing, pushing until it contacts the solvent tubing. Solvent filter PTFE tubing Solvent tubing Insert the filtered end of the solvent tubing into the solvent bottle. Shake the filter to remove any air from it. Prime the binary solvent manager.
Page 227: Replacing The Air Filters In The Binary Solvent Manager Door
To clean the air filters Using the T10 TORX driver, remove the 8 screws that secure the air filter frames and air filters to the inside of the binary solvent manager door. Air filter Air filter Frame Frame TP02429 Remove the air filters from the air filter frames and use a vacuum to clean them.
Page 228: Removing And Replacing The Primary Head Seals
8 screws. Removing and replacing the primary head seals See the ACQUITY UPLC Console online Help to help determine whether you need to replace the primary head seals. To prevent injury, always observe Good Laboratory...
Page 229 • Plunger (recommended) To remove the primary head Flush the binary solvent manager with non-hazardous solvent. In the ACQUITY UPLC Console, select Binary Solvent Manager from the system tree. In the binary solvent manager information window, click Maintain > Heads.
Page 230 Power-off the binary solvent manager. The binary solvent manager is referred to as “pump” on the warning Tip: label affixed to the i Valve actuator. To avoid solvent spills, move the solvent bottles to a Warning: location below the binary solvent manager. Move the solvent bottles to a location below the binary solvent manager.
Page 231 Loosen the cap nut on the in-line filter so that it is all the way off the ferrule holder fitting threads. Cap nut TP02958 Ferrule holder fitting Cap nut Maintaining the binary solvent manager 6-55...
Page 232 10. Use the 5/16-inch open-end wrench to loosen the shell nut and then fully unscrew it. Shell nut 5/16-inch wrench Caution: • When you remove the valve assembly, ensure the PEEK washer, which is normally on the top face of the i Valve cartridge, does not remain in the head.
Page 233 11. Remove the i Valve actuator from the bottom of the primary pump head. Bottom of primary pump head Valve actuator 12. Remove both seal wash tubes secured to the seal wash housing by barbed fittings, using a tool or by pulling on the tubes as close to the head as possible.
Page 234 13. Using a pliers, remove the drip wire from the head assembly. Drip wire Pliers 14. Use the 1/4-inch open-end wrench to disconnect the outlet tube from the transducer. Transducer Outlet tube TP02966 1/4-inch open-end wrench 6-58 Maintaining the System...
Page 235 15. Disconnect the pressure transducer cable from the bulkhead by squeezing on the tabs and pulling gently. Tabs Pressure transducer cable connector 16. Using the T27 TORX driver, loosen the 2 head bolts 1/2-turn. The bolts are accessible from the front of the pressure transducer. Head bolt access hole Head bolt...
Page 236 Removing support plate bolts Support plate bolt (2) T27 TORX driver Pulling head and support plate off actuator housing Plunger Head and support plate 6-60 Maintaining the System...
Page 237 To remove the primary head seals Stand the head upright on a clean surface. Using the T27 TORX driver, completely loosen the 2 head bolts to release the support plate from the pump head. T27 TORX driver Head bolt access hole Lift the pump head from the support plate.
Page 238 Head seals Head PTFE O-ring Seal wash housing Head seal Seal wash seal Seal wash spacer Using the smooth end of the seal extraction tool, pull the seal wash spacer from the head. Seal wash spacer Seal extraction tool 6-62 Maintaining the System...
Page 239 Taking care not to scratch any surfaces, screw the threaded end of the seal extraction tool into the head seal and carefully pull it out of the head. Head seal TP03001 Seal extraction tool Taking care not to scratch any metal surfaces, use a sharp tool to remove the PTFE O-ring.
Page 240 Lubricate the new head seal with methanol, and use the smooth end of the seal extraction tool to place it in the head. Head seal Seal extraction tool TP02971 Seal extraction tool Head seal 6-64 Maintaining the System...
Page 241 10. Center the seal wash spacer over the head seal so that the cross-side faces upward. Seal wash spacer with cross-side facing up 11. Orient the seal wash housing so that the holes on the side of it line up with the holes on the side of the head, and then drop it into place.
Page 242 14. Holding the assembly together, use the T27 TORX driver to tighten the 2 head bolts finger-tight. T27 TORX driver Head bolt Head bolt access hole access hole Replace the plunger whenever you replace the head Recommendation: seal. See “To replace the primary head plunger” on page 6-97.
Page 243 Hold the head assembly securely against the actuator housing, and then use the T27 TORX driver to tighten the support plate screws securely. Support plate bolt hole Head bolt access hole (2) Support plate bolt T27 TORX driver Tighten the head bolts evenly as well. Reinstall the drip wire around the head assembly, ensuring that the tip is in the 6 o’clock position.
Page 244 condition. Use the 5/16-inch open-end wrench to tighten the nut an additional 1/8-turn. Shell nut 5/16-inch wrench Reinsert the in-line filter and tube into the ferrule holder fitting. Ferrule holder fitting In-line filter 6-68 Maintaining the System...
Page 245 10. Place the cap nut over the ferrule holder fitting and finger-tighten the cap nut to the extent possible. Ferrule holder fitting Cap nut 11. Align the red dot on the i Valve connector with the red dot on the receptacle, in the 12 o’clock position, and insert the connector into the receptacle.
Page 246: Removing And Replacing The Accumulator Head Seals
Removing and replacing the accumulator head seals See the ACQUITY UPLC Console online Help to help determine whether you need to replace the accumulator head seals. To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the binary solvent manager.
Page 247 In the Head Maintenance dialog box, select the head (Accumulator) from the side of the solvent manager (A or B) that you plan to perform maintenance on. Click Move Backward, and then wait for the plunger to stop. To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to an instrument.
Page 248 Using pliers, remove the drip wire from the head assembly. Drip wire Pliers 10. Use the 1/4-inch open-end wrench to disconnect the outlet tube from the transducer. Transducer Outlet tube TP03030 1/4-inch open-end wrench 6-72 Maintaining the System...
Page 249 11. Using the 5/16-inch open-end wrench to hold the check valve cartridge in place, disconnect the tubing connection from the check valve with the 1/4-inch open-end wrench. 5/16-inch open-end wrench Check valve Tubing connection 1/4-inch open-end wrench TP02433 12. Disconnect the pressure transducer cable from the bulkhead by squeezing on the tabs and pulling gently.
Page 250 13. Using the T27 TORX driver, loosen the 2 head bolts 1/2-turn. The bolts are accessible from the front of the pressure transducer. Head bolt access hole Head bolt access hole T27 TORX driver To avoid damaging the plunger, support the head from Caution: below as you remove it.
Page 251 Removing support plate bolts Support plate bolt (2) T27 TORX driver Pulling head and support plate off actuator housing Plunger Head and support plate Maintaining the binary solvent manager 6-75...
Page 252 To remove the accumulator head seals Stand the head upright on a clean surface. Using the T27 TORX driver, completely loosen the 2 head bolts to release the support plate from the pump head. T27 TORX driver Head bolt access hole TP02989 Lift the pump head from the support plate.
Page 253 Remove the old seal wash seal and discard it. Head seals Head PTFE O-ring Seal wash housing Head seal Seal wash seal Seal wash spacer Using the smooth end of the seal extraction tool, pull the seal wash spacer from the head. Seal wash spacer Seal extraction tool Maintaining the binary solvent manager...
Page 254 Taking care not to scratch any surfaces, screw the threaded end of the seal extraction tool into the head seal and carefully pull it out of the head. Head seal TP03001 Seal extraction tool Taking care not to scratch any metal surfaces, use a sharp tool to remove the PTFE O-ring.
Page 255 Lubricate the new head seal with methanol, and use the smooth end of the seal extraction tool to place it in the head. Head seal Seal extraction tool TP02971 Seal extraction tool Head seal Maintaining the binary solvent manager 6-79...
Page 256 10. Center the seal wash spacer over the head seal so that the cross-side faces upward. Seal wash spacer with cross-side facing up 11. Orient the seal wash housing so that the holes on the side of it line up with the holes on the side of the head, and then drop it into place.
Page 257 14. Holding the assembly together, use the T27 TORX driver to tighten the 2 head bolts finger-tight. T27 TORX driver Head bolt access holes Replace the plunger whenever you replace the head Recommendation: seal. See “To replace the accumulator head plunger” on page 6-114.
Page 258 Hold the head assembly securely against the actuator housing, and then use the T27 TORX driver to tighten the support plate screws securely. Support plate bolt hole Support plate bolt T27 TORX driver Tighten the head bolts evenly as well. Reinstall the drip wire around the head assembly, ensuring that the tip is in the 6 o’clock position.
Page 259: Replacing The Primary Head Plungers
Sharp tool (recommended) To remove the primary head Flush the binary solvent manager with non-hazardous solvent. In the ACQUITY UPLC Console, select Binary Solvent Manager from the system tree. In the binary solvent manager information window, click Maintain > Heads.
Page 260 In the Head Maintenance dialog box, select the head (Primary) from the side of the solvent manager (A or B) that you plan to perform maintenance on. Click Move Backward, and then wait for the plunger to stop. To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to an instrument.
Page 261 Grasp the i Valve connector by the knurled diameter and pull it toward you to disconnect it from the receptacle. Connector receptacle Valve connector Knurled diameter Maintaining the binary solvent manager 6-85...
Page 262 Loosen the cap nut on the in-line filter so that it is all the way off the ferrule holder fitting threads. Cap nut TP02958 Ferrule holder fitting Cap nut 6-86 Maintaining the System...
Page 263 10. Use the 5/16-inch open-end wrench to loosen the shell nut and then fully unscrew it. Shell nut 5/16-inch wrench Caution: • When you remove the valve assembly, ensure the PEEK washer, which is normally on the top face of the i Valve cartridge, does not remain in the head.
Page 264 11. Remove the i Valve actuator from the bottom of the primary pump head. Bottom of primary pump head Valve actuator 12. Remove both seal wash tubes secured to the seal wash housing by barbed fittings, using a tool or by pulling on the tubes as close to the head as possible.
Page 265 13. Using pliers, remove the drip wire from the head assembly. Drip wire Pliers 14. Use the 1/4-inch open-end wrench to disconnect the outlet tube from the transducer. Transducer Outlet tube TP02966 1/4-inch open-end wrench Maintaining the binary solvent manager 6-89...
Page 266 15. Disconnect the pressure transducer cable from the bulkhead by squeezing on the tabs and pulling gently. Tabs Pressure transducer cable connector 16. Using the T27 TORX driver, loosen the 2 head bolts 1/2-turn. The bolts are accessible from the front of the pressure transducer. Head bolt access hole Head bolt...
Page 267 Pulling head and support plate off actuator housing Plunger Head and support plate Waters strongly recommends that you replace the Recommendation: head seals when you replace the plunger. If you are not replacing any seals as part of this procedure, proceed to “To replace the primary head...
Page 268 To remove the primary head seals Stand the head upright on a clean surface. Using the T27 TORX driver, completely loosen the 2 head bolts to release the support plate from the pump head. T27 TORX driver Head bolt access hole Lift the pump head from the support plate.
Page 269 Head seals Head PTFE O-ring Seal wash housing Head seal Seal wash seal Seal wash spacer Using the smooth end of the seal extraction tool, pull the seal wash spacer from the head. Seal wash spacer Seal extraction tool Maintaining the binary solvent manager 6-93...
Page 270 Taking care not to scratch any surfaces, screw the threaded end of the seal extraction tool into the head seal and carefully pull it out of the head. Head seal TP03001 Seal extraction tool Taking care not to scratch any metal surfaces, use a sharp tool to remove the PTFE O-ring.
Page 271 Lubricate the new head seal with methanol, and use the smooth end of the seal extraction tool to place it in the head. Head seal Seal extraction tool TP02971 Seal extraction tool Head seal Maintaining the binary solvent manager 6-95...
Page 272 10. Center the seal wash spacer over the head seal so that the cross-side faces upward. Seal wash spacer with cross-side facing up 11. Orient the seal wash housing so that the holes on the side of it line up with the holes on the side of the head, and then drop it into place.
Page 273 To replace the primary head plunger Stand the head assembly upright on a clean surface, and set it aside. Waters strongly recommends that you replace the Recommendation: head seals when you replace the plunger. Maintaining the binary solvent manager...
Page 274 Use the recessed side of the plunger removal tool to apply pressure to both sides of the release collar, and then remove the old plunger. Recessed lip of plunger removal tool TP02990 Spring-loaded release collar Plunger Plunger removal tool While applying pressure to the release collar with the plunger removal tool, grasp the new plunger with a clean, lint-free cloth, and insert it.
Page 275 To reattach the primary head Lubricate the seals and plunger with methanol. To avoid damaging the plunger, ensure that the head Caution: assembly is not tilted relative to the plunger when you position it on the mechanism. Carefully slide the head assembly and support plate over the sapphire plunger, making sure not to tilt the head.
Page 276 Connect the pressure transducer cable to the bulkhead. Pressure transducer cable connector Orient the i Valve assembly so that the cable exits from the left-hand side. Insert the i Valve assembly into the bottom of the primary pump head, and route the cable behind the valve actuator. Finger-tighten the shell nut to secure the valve.
Page 277 Reinsert the in-line filter and tube into the ferrule holder fitting. Ferrule holder fitting In-line filter 10. Place the cap nut over the ferrule holder fitting and finger-tighten the cap nut to the extent possible. Ferrule holder fitting Cap nut Maintaining the binary solvent manager 6-101...
Page 278 11. Align the red dot on the i Valve connector with the red dot on the receptacle, in the 12 o’clock position, and insert the connector into the receptacle. Red dot on connector Red dot on receptacle Connector receptacle Valve connector Knurled diameter 12.
Page 279: Replacing The Accumulator Head Plungers
Sharp tool (recommended) To remove the accumulator head Flush the binary solvent manager with non-hazardous solvent. In the ACQUITY UPLC Console, select Binary Solvent Manager from the system tree. In the binary solvent manager information window, click Maintain > Heads.
Page 280 In the Head Maintenance dialog box, select the head (Accumulator) from the side of the solvent manager (A or B) that you plan to perform maintenance on. Click Move Backward, and then wait for the plunger to stop. To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to an instrument.
Page 281 Using pliers, remove the drip wire from the head assembly. Drip wire Pliers 10. Use the 1/4-inch open-end wrench to disconnect the outlet tube from the transducer. Transducer Outlet tube TP03030 1/4-inch open-end wrench Maintaining the binary solvent manager 6-105...
Page 282 11. Using the 5/16-inch open-end wrench to hold the check valve cartridge in place, disconnect the tubing connection from the check valve with the 1/4-inch open-end wrench. 5/16-inch open-end wrench Check valve Tubing connection 1/4-inch open-end wrench TP02433 12. Disconnect the pressure transducer cable from the bulkhead by squeezing on the tabs and pulling gently.
Page 283 13. Using the T27 TORX driver, loosen the 2 head bolts 1/2-turn. The bolts are accessible from the front of the pressure transducer. Head bolt access hole Head bolt access hole T27 TORX driver To avoid damaging the plunger, support the head from Caution: below as you remove it.
Page 284 Pulling head and support plate off actuator housing Plunger Head and support plate Waters strongly recommends that you replace the Recommendation: head seals when you replace the plunger. If you are not replacing any seals as part of this procedure, proceed to “To replace the accumulator...
Page 285 To remove the accumulator head seals Stand the head upright on a clean surface. Using the T27 TORX driver, completely loosen the 2 head bolts to release the support plate from the pump head. T27 TORX driver Head bolt access hole Lift the pump head from the support plate.
Page 286 Head seals Head PTFE O-ring Seal wash housing Head seal Seal wash seal Seal wash spacer Using the smooth end of the seal extraction tool, pull the seal wash spacer from the head. Seal wash spacer Seal extraction tool 6-110 Maintaining the System...
Page 287 Taking care not to scratch any surfaces, screw the threaded end of the seal extraction tool into the head seal and carefully pull it out of the head. Head seal TP03001 Seal extraction tool Taking care not to scratch any metal surfaces, use a sharp tool to remove the PTFE O-ring.
Page 288 Lubricate the new head seal with methanol, and use the smooth end of the seal extraction tool to place it in the head. Head seal Seal extraction tool TP02971 Seal extraction tool Head seal 6-112 Maintaining the System...
Page 289 10. Center the seal wash spacer over the head seal so that the cross-side faces upward. Seal wash spacer with cross-side facing up 11. Orient the seal wash housing so that the holes on the side of it line up with the holes on the side of the head, and then drop it into place.
Page 290 Head bolt access holes To replace the accumulator head plunger Stand the head assembly upright on a clean surface, and set it aside. Waters strongly recommends that you replace the Recommendation: head seals when you replace the plunger. 6-114 Maintaining the System...
Page 291 Use the recessed side of the plunger removal tool to apply pressure to both sides of the release collar, and then remove the old plunger. Recessed lip of plunger removal tool TP02990 Spring-loaded release collar Plunger Plunger removal tool While applying pressure to the release collar with the plunger removal tool, grasp the new plunger with a clean, lint-free cloth, and insert it.
Page 292 To reattach the accumulator head Lubricate the seals and plunger with methanol. To avoid damaging the plunger, ensure that the head Caution: assembly is not tilted relative to the plunger when you position it on the mechanism. Carefully slide the head assembly and support plate over the sapphire plunger, making sure not to tilt the head.
Page 293: Replacing The Vent Valve Cartridge
Connect the pressure transducer cable to the bulkhead. Pressure transducer cable connector Reattach all fittings and seal wash tubes. Return the solvent bottles to their original location. Power-on the binary solvent manager. 10. Prime the binary solvent manager (see “Priming the binary solvent manager”...
Page 294 To replace the vent valve cartridge In the ACQUITY UPLC Console, select Binary Solvent Manager from the system tree. In the binary solvent manager information window, click Interactive Display. In the binary solvent manager interactive display dialog box, click Control Ensure the vent valve is set to Vent.
Page 295 The vent valve cartridge must slide fully into the vent Requirement: valve assembly. If it does not, contact Waters Technical Service. 11. Insert the 2-mm hex screw at the 10 o’clock position on the vent valve cartridge. Use the 2-mm Allen wrench to tighten it.
Page 296: Replacing The Fuses
If your binary solvent manager has the power entry module shown below and it fails to power-on, the status LEDs are unlit, or the power supply fan does not operate, call Waters Technical Service (see page 6-6).
Page 297 Required materials • 5 A, 250 V, 5 × 20 mm, slow-blow, IEC type fuse (2) • Flat-blade screwdriver To replace the fuses Replace both fuses, even when only one is defective. Requirement: To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to an instrument.
Page 298: Cleaning The Instrument's Exterior
Insert the new fuses into the holders and the holders into the power entry module. Reconnect the power cord to the power entry module. Cleaning the instrument’s exterior Use a soft cloth, dampened with water, to clean the outside of the binary solvent manager.
Page 299: Resolving Sample Manager Leak Sensor Errors
In the Defrost dialog box, click Start. The sample compartment defrosts for 60 minutes. Resolving sample manager leak sensor errors The sample manager is the only ACQUITY UPLC instrument with 2 leak sensors, bottom and top, called the sample manager leak sensor and the column heater leak sensor, respectively.
Page 300 To resolve a sample manager leak sensor error View the Leak Sensors dialog box in the ACQUITY UPLC Console to determine which of the sample manager’s 2 leak sensors detected a leak. If the message reads “Leak Detected”, slide out the sample manager fluidics tray.
Page 301 Use a nonabrasive, lint-free wipe to dry the leak sensor prism. Prism Lint-free wipe TP02891 Roll up a nonabrasive, lint-free wipe, and use it to absorb the liquid from the leak sensor reservoir and its surrounding area. Leak sensor reservoir Rolled up lint-free wipe Maintaining the sample manager 6-125...
Page 302 With a cotton swab, absorb any remaining liquid from the corners of the leak sensor reservoir and its surrounding area. Leak sensor reservoir Cotton swab 6-126 Maintaining the System...
Page 303 If you detached the connector from the front of the instrument, reattach 10. In the ACQUITY UPLC Console, select Sample Manager from the system tree. 11. In the sample manager information window, click Control > Reset SM, to reset the sample manager.
Page 304: Replacing The Sample Manager's Leak Sensor
Replacing the sample manager’s leak sensor The leak sensor can be contaminated with biohazardous Warning: and/or toxic materials. Always wear clean, chemical-resistant, powder-free gloves when performing this procedure. Required materials • Clean, chemical-resistant, powder-free gloves • New leak sensor To replace the sample manager’s leak sensor Open the sample manager door, gently pulling its right-hand edge toward you.
Page 305 Remove the leak sensor from its reservoir by grasping it by its serrations and pulling upward on it. Serrations Unpack the new leak sensor. Maintaining the sample manager 6-129...
Page 306 Plug the leak sensor connector into the front of the instrument. In the ACQUITY UPLC Console, select Sample Manager from the system tree. In the sample manager information window, click Control > Reset SM, to reset the sample manager.
Page 307: Replacing The Sample Needle Assembly
Required material Needle assembly To remove the needle assembly In the ACQUITY UPLC Console, click Sample Manager, in the left-hand pane. Select Maintain > Change needle. A message requesting that you remove the right-hand sample plate from the sample manager compartment appears.
Page 308 Unscrew the needle assembly’s fitting from injector port 3. Port 3 fitting Remove the fitting from the needle. In the sample compartment, loosen the red thumbscrew on the needle mounting bracket. Extended puncture needles have a grey plastic or stainless steel Tip: thumbscrew on the needle mounting bracket.
Page 309 XYZ Mechanism Needle latch Red thumbscrew (grey plastic or stainless steel on extended puncture needle assemblies) Push the needle latch back to release the needle mounting cylinder from its mounting cavity. Flag Mounting cavity Needle latch Mounting cylinder Loose loop of tubing Maintaining the sample manager 6-133...
Page 310 To avoid puncture wounds, or damage to the end of the Warning: needle, do not touch or press the end of the sample needle. 10. Remove the needle assembly from the sample compartment. ACQUITY UPLC sample needle assembly Fitting ID sleeve Protective cap...
Page 311 To install the sample needle assembly Insert the needle tip into the guide tube next to the injector. Needle guide tube Needle assembly tube Gently push the needle assembly into the sample compartment. From inside the sample compartment, take hold of the needle assembly tubing as it enters the chamber from above.
Page 312 Insert the needle mounting cylinder into the mounting cavity. Flag Mounting cavity Needle latch Mounting cylinder Loose loop of tubing Pull the needle latch forward to secure the needle assembly. Form a loop so that the portion of the tubing with the black clamping sleeve can be secured by the thumbscrew.
Page 313 Route the tubing through the groove below the Z-flag, as shown in the figure below. Z-flag Groove Loose loop of tubing Incorrect placement of loose loop of tubing Loose loop of tubing Z-flag Groove 10. Align the tubing so that the portion just to the left-hand side of the clamping sleeve fits into the notch behind the thumbscrew.
Page 314 13. Close the sample manager door. 14. On the ACQUITY UPLC Console, click Control > Prime syringe. 15. In the Prime Syringes dialog box, select Sample syringe and wash syringes.
Page 315: Calibrating The Needle Z-Axis
Calibrating the needle Z-axis You must calibrate the needle Z-axis whenever you replace the sample needle. To calibrate the needle Z-axis Click Maintain > Calibrate needle Z axis. In the Calibrate Needle Z Axis dialog box, click Start, and then click OK in the confirmation window.
Page 316: Characterizing The Needle Seal
If characterization is unsuccessful, examine the needle to ensure you installed it properly. Make any necessary adjustments, and then characterize the needle seal again. If unsuccessful, consult the ACQUITY UPLC Console online Help. If the characterization is successful, click Close. Characterizing the needle and sample loop volumes Perform this procedure whenever you change the •...
Page 317 • sample loop. • needle. • syringes. Whenever you replace the sample loop and/or sample needle, you must characterize the volume of the replacement parts to accommodate the differently sized parts and to ensure optimal chromatographic results. Perform this procedure regardless of whether the sizes of the replacement parts equal those of the original parts or differ from them.
Page 318: Replacing The Puncture Needle Holder
1/4-turn counterclockwise, releasing the tray. Remove the well plate tray from the sample manager compartment. In the ACQUITY UPLC Console, click OK to move the needle to the maintenance position. 6-142 Maintaining the System...
Page 319 Place a sheet of paper on the floor of the sample manager compartment, beneath the puncture needle. The paper will catch the needle if it falls during replacement. Remove the needle clean system tube from the barbed fitting on the existing puncture needle holder.
Page 320 10. Slide the puncture needle holder over the sample needle and into the housing. Orient the needle so that the barbed fitting enters the slot on the needle block. Needle block Set screw Barbed fitting Slot 11. Hold the puncture needle assembly in the fully up position while you tighten the set screw with the T6 TORX driver.
Page 321: Replacing The Sample Loop
Required materials • 1/4-inch open-end wrench • ACQUITY UPLC sample loop To replace the sample loop Slide out the sample manager fluidics tray. To avoid kinks, do not pull on the volume detection Caution: device (port 2) or sample needle (port 3) tubing.
Page 322 Sample loop fittings on the injection valve Port 1 fitting Outlet to column Port 4 fitting Inlet from pump Sample loop Remove the sample loop and its fittings. If you are going to reuse the sample loop, label each end with the port it was connected to.
Page 323 • Use only Waters approved ACQUITY UPLC loops with ACQUITY UPLC fittings. Each loop must be fitted to its injector. Ensure the 2-piece ferrule is in place on one end of the sample loop. If not, slide a 2-piece ferrule and compression screw onto one end of the sample loop, and then seat the end into injection valve port 1.
Page 324 15. Slide the fluidics tray closed. 16. Ensure the sample manager compartment door is closed. 17. In the ACQUITY UPLC Console, select Sample Manager from the system tree. 18. Click Maintain > Calibrate System Volume to characterize the new system volumes.
Page 325: Replacing The Sample Syringe
Replacing the sample syringe Air bubbles in the syringe adversely affect system pressure, baseline, volume, and peak area. Remove air bubbles by gently tapping on the syringe as its plunger retracts, such as when priming the sample syringe. Replace the sample syringe when any of these conditions arise: •...
Page 326 The 50-μL loop must be used with a 250-μL sample syringe. To replace the sample syringe In the ACQUITY UPLC Console, select Sample Manager from the system tree. Click Maintain > Replace > Sample syringe. Doing so invokes a wizard that moves the syringe to the down position.
Page 327 Unscrew the sample syringe counterclockwise until it separates from the sample syringe valve. To avoid breaking the syringe, do not grasp it by its Caution: glass barrel. Always grasp the syringe by its knurled collar. Depress the syringe barrel to clear the top mounting bracket, and remove the syringe.
Page 328: Replacing The Wash Syringes
Replacing the wash syringes The only wash syringe size available is 2.5 mL. To ensure accurate results, use only syringes that are approved by Waters for use in the ACQUITY UPLC system. Replace the wash syringes when the syringe plunger’s tip leaks, becomes worn or discolored, or fails the leak test.
Page 329 Wash syringe assembly components Mounting bracket Weak wash syringe Strong wash syringe Mounting bracket Knurled screws Looking down at the wash syringes, unscrew each syringe, clockwise, until it separates from the mounting bracket. To avoid breaking the syringe, do not grasp it by its Caution: glass barrel.
Page 330 Depress the syringe barrel to clear the top mounting bracket, and remove the syringe. Knurled collar Mounting bracket Unpack each replacement wash syringe. Partially fill each new wash syringe (by hand) with weak or strong wash solvent (to help remove air bubbles). Pull each wash syringe plunger down so that the plunger end slides over the threaded post on the syringe guide mounting bracket.
Page 331: Modifying Sample Syringe Configuration Parameters
Modifying sample syringe configuration parameters To configure the system for a syringe size that is different from the one currently fitted In the ACQUITY UPLC Console, select Sample Manager from the system tree. Select Configure > Volumes. In the Volume Configuration dialog box, select the appropriate sample syringe size from the list, and then click OK.
Page 332: Replacing The Injection Valve Cartridge
Injection valve cartridge • Sample loop To replace the injection valve cartridge In the ACQUITY UPLC Console, select Sample Manager from the system tree. In the sample manager information window, click Maintain > Park Inject Valve. To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to an instrument.
Page 333 Remove the fittings attached to the injection valve cartridge. Fittings Fittings Use the 2-mm Allen wrench to remove the screw at the 10 o’clock position on the injection valve cartridge. Remove the injection valve cartridge from the injection valve assembly. Carefully unpack the replacement injection valve cartridge.
Page 334 If the injection valve cartridge does not slide fully into the Requirement: injection valve assembly, contact your Waters service representative. 12. Insert the 2-mm hex screw at the 10 o’clock position on the injection valve cartridge, and use the 2-mm Allen wrench to tighten it.
Page 335: Testing The Sample Manager Backpressure Regulator
18. In the ACQUITY UPLC Console, select Sample Manager from the system tree. 19. In the sample manager information window, click Control > Prime syringes. 20. In the Prime Syringes dialog box select sample syringe and wash syringes. type 1 in the Number of Cycles text box.
Page 336 Power entry module on recently manufactured instruments To avoid electric shock, power-off and unplug the sample Warning: manager before examining fuses. For continued protection against fire, replace fuses with those of the same type and rating only. Suspect an open or otherwise defective fuse when any of these conditions apply: •...
Page 337: Cleaning The Instrument's Exterior
Disconnect the power cord from the power entry module. Use a flat-blade screwdriver to open the fuse holder door, located above the power entry module on the rear panel. With minimum pressure, pull on each spring-loaded fuse holder and remove it. Fuse holder door Fuse Fuse holders...
Page 338: Maintaining The Ht Column Heater
Nonabrasive, lint-free wipes To resolve a HT column heater leak sensor error View the Leak Sensors dialog box in the ACQUITY UPLC Console to determine which of the sample manager’s 2 leak sensors detected a leak. If the message reads “Leak Detected (Column)”, open the HT column heater’s door.
Page 339 Locate the source of the leak, and make the repairs necessary to stop the leak. Liquid from a detector leak can accumulate in the HT column Tip: heater leak sensor reservoir and cause the HT column heater leak sensor alarm to sound. Always inspect the detector when attempting to locate the source of a leak.
Page 340 Remove the leak sensor from its reservoir by grasping it by its serrations, pulling upward on it, and tilting it to the left-hand side. Serrations If you cannot easily manipulate the leak sensor after removing it Tip: from its reservoir, detach the connector from the front of the instrument (see page 6-171).
Page 341 Roll up a nonabrasive, lint-free wipe, and use it to absorb the liquid from the leak sensor reservoir and its surrounding area. Leak sensor reservoir Rolled up lint-free wipe 10. With a cotton swab, absorb any remaining liquid from the corners of the leak sensor reservoir and its surrounding area.
Page 342 12. If you detached the connector from the front of the instrument, reattach 13. In the ACQUITY UPLC Console, select Sample Manager from the system tree. 14. In the sample manager information window, click Control > Reset SM, to reset the sample manager.
Page 343: Resolving Ht Column Heater Leak Sensor Errors
To resolve a leak sensor error for a HT column heater with its door fully opened View the Leak Sensors dialog box in the ACQUITY UPLC Console to determine which of the sample manager’s 2 leak sensors detected a leak.
Page 344 Slide out the sample manager fluidics tray. To avoid damaging the leak sensor, do not grasp it by Caution: the ribbon cable. Using one finger from each hand, remove the leak sensor from its reservoir by grasping it by its serrations and pulling upward on it. Serrations Leave the leak sensor in place, use the syringe included the Alternative:...
Page 345 Tilt the leak sensor to the right-hand side and remove it from under the drip tray. Leak sensor tilted to right If you cannot easily manipulate the leak sensor after removing it Tip: from its reservoir, detach the connector from the front of the instrument (see page 6-171).
Page 346 If you detached the connector from the front of the instrument, reattach 10. Slide the fluidics tray closed. 11. In the ACQUITY UPLC Console, select Sample Manager from the system tree. 12. In the sample manager information window, click Control > Reset SM, to reset the sample manager.
Page 347: Replacing The Ht Column Heater's Leak Sensor
Replacing the HT column heater’s leak sensor The leak sensor can be contaminated with biohazardous Warning: and/or toxic materials. Always wear clean, chemical-resistant, powder-free gloves when performing this procedure. Required materials • Clean, chemical-resistant, powder-free gloves • New leak sensor To replace the HT column heater’s leak sensor Open the sample manager door, gently pulling its right-hand edge toward you.
Page 348 Push down on the metal clip at the bottom, left-hand side of the HT column heater to release the door, and then pull the door toward you. Column heater door Metal clip TP02596 Swing the door fully to the right-hand side. Column heater door Remove the leak sensor from its reservoir by grasping it by its...
Page 349 10. Plug the leak sensor connector into the front of the instrument. 11. In the ACQUITY UPLC Console, select Sample Manager from the system tree. 12. In the sample manager information window, click Control > Reset SM, to reset the sample manager.
Page 350: Replacing The Ht Column Heater's Leak Sensor (Door Fully Opened)
Replacing the HT column heater’s leak sensor (door fully opened) The leak sensor can be contaminated with biohazardous Warning: and/or toxic materials. Always wear clean, chemical-resistant, powder-free gloves when performing this procedure. Required materials • Leak sensor • Clean, chemical-resistant, powder-free gloves To replace the leak sensor for a HT column heater with its door fully opened Open the sample manager door, gently pulling its right-hand edge toward you.
Page 351 Using one finger from each hand, remove the leak sensor from its reservoir by grasping it by its serrations and pulling upward on it. Serrations Tilt the leak sensor to the right-hand side and remove it from under the drip tray. Leak sensor tilted to right Unpack the new leak sensor.
Page 352: Replacing The Column In The Ht Column Heater
Slide the fluidics tray closed. 10. In the ACQUITY UPLC Console, select Sample Manager from the system tree. 11. In the sample manager information window, click Control > Reset SM, to reset the sample manager. Replacing the column in the HT column heater...
Page 353: Replacing The Column Stabilizer Assembly In The Ht Column Heater
Replace the ferrule. Continue with “Installing the column in an HT column heater” on page 2-46. Replacing the column stabilizer assembly in the HT column heater To prevent burn injuries, set the column temperature to Off, and Warning: then allow the column compartment and its components to cool for 60 minutes before touching them.
Page 354: Replacing The Frit In The Ht Column Heater's In-Line Filter Unit
Replacing the frit in the HT column heater’s in-line filter unit If your system has the optional column in-line filter installed, follow the instructions below to replace the frit. To prevent contamination, wear clean, chemical-resistant, Caution: powder-free gloves when replacing the frit in the column in-line filter unit. Because of wear on the non-replaceable compression Recommendation: fittings, replace the column in-line filter unit after you use the six frits that...
Page 355 Column components for HT column heater Flow In-line filter unit From column stabilizer Column tubing assembly Inlet TP02873 Reusable fitting Detach the column stabilizer tubing assembly from the column in-line filter unit. Do not detach the column in-line filter unit from the column inlet.
Page 356: Maintaining The Column Manager And Column Heater/Cooler
Start the mobile phase and examine the assembled components for leaks. Tighten fittings as needed. Return the assembled components to the column tray, push down the metal latch to secure the tubing, and tilt the tray to the closed position. Maintaining the column manager and column heater/cooler Under normal operating conditions, the column manager and column...
Page 357 To resolve a column manager or column heater/cooler leak sensor error View the Leak Sensors dialog box in the ACQUITY UPLC Console to confirm that the column manager leak sensor detected a leak. Locate the source of the leak, and make the repairs necessary to stop the leak.
Page 358 Roll up a nonabrasive, lint-free wipe, and use it to absorb the liquid from the leak sensor reservoir and its surrounding area. Rolled up lint-free wipe Leak sensor reservoir With a cotton swab, absorb any remaining liquid from the corners of the leak sensor reservoir and its surrounding area.
Page 359 Align the leak sensor’s T-bar with the slot in the side of the leak sensor reservoir, and slide the leak sensor into place. TP02892 T-bar TP02919 Slot in leak sensor reservoir Leak sensor installed in reservoir Ensure the ribbon cable is routed behind the cable clip. Cable clip Ribbon cable Maintaining the column manager and column heater/cooler 6-183...
Page 360: Replacing The Column Manager's Or Column Heater/Cooler's Leak Sensor
If you detached the connector from the front of the instrument, reattach 10. In the ACQUITY UPLC Console, select Column Manager from the system tree. 11. In the column manager information window, click Control > Reset CM, to reset the column manager.
Page 361 Remove the leak sensor from its reservoir by grasping it by its serrations and pulling upward on it. Serrations Unpack the new leak sensor. Align the leak sensor’s T-bar with the slot in the side of the leak sensor reservoir, and slide the leak sensor into place. TP02892 T-bar TP02919...
Page 362: Replacing The Column In The Column Manager
Cable clip Ribbon cable Plug the leak sensor connector into the front of the instrument. In the ACQUITY UPLC Console, select Column Manager from the system tree. In the column manager information window, click Control > Reset CM, to reset the column manager.
Page 363 (10 to 30 °C [50 to 86 °F]) and allow the unit to attain this set-point temperature. In the ACQUITY UPLC Console, click the Stop Flow button, and wait until all flow is stopped. Maintaining the column manager and column heater/cooler 6-187...
Page 364 Open the door and locate the column you want to replace. Column number 1 is at the top position, and column number 4 is at the bottom. Swing the wire bail open. Wire bail To disconnect the outlet side of the column from the post-cooler, pull the left-hand end of the column (outlet to detector) and the U-shaped, 6-188 Maintaining the System...
Page 365 left-hand column clip away from the column slot. Unscrew the finger-tight PK fitting carefully from the column end. Column clip If removal of the finger-tight PK fitting requires more force, place the 5/16-inch wrench on the end of the column immediately to the right-hand side of the column clip to hold the column steady, and loosen the PK fitting with the PK fitting wrench.
Page 366 Disconnecting the outlet side of the top column Finger-tight PK fitting PEEK tube To disconnect the inlet side of the column from the preheater, pull the right-hand end of the column (inlet from preheater) and the right-hand column clip away from the column slot. Holding the gold compression screw steady, unscrew the right-hand end of the column to disconnect it from the preheater on the right-hand side.
Page 367 Detach the eCord from its port on the right-hand side of the column manager. The eCord LED is constant green when the eCord is connected, and Tip: darkens when you detach the eCord from the port. Verify that the darkened LED corresponds to the column that you removed. eCord fob Column 1 LED Tether...
Page 368 You can install up to four ACQUITY UPLC (or similar) columns in Tip: the column manager or column heater/cooler. To install a column Carefully connect the inlet end of the column (pressure up to 103,421 kPa [1034 bar, 15,000 psi]) to the preheater. Use a gold compression screw, PEEK ferrule, and stainless steel collet on the stainless steel tube.
Page 369: Replacing The Column In The Column Heater/Cooler
• If the column has no eCord, you can add a comment in the ACQUITY UPLC Console to identify the column. Replacing the column in the column heater/cooler To prevent burn injuries, adjust the temperature set-point of...
Page 370 To remove the column Stop any currently running mobile phase before replacing the column. After cooling, swing the column retainer rod out to the right-hand side. Column retainer rod Disconnect the stabilizer outlet tube from the column or in-line filter. Place the forked end of the collet separator over the collet on the column stabilizer tube, and gently wiggle it back and forth to pry the reusable fitting loose from the collet.
Page 371: Replacing The Column Stabilizer Assembly In The Column Heater/Cooler
Replacing the column stabilizer assembly in the column heater/cooler To prevent burn injuries, adjust the temperature set-point of Warning: the column manager or column heater/cooler to a safe temperature (10 to 30 °C [50 to 86 °F]) and allow the unit to attain this set-point temperature prior to handling components within the column compartment.
Page 372: Replacing The Frit In The Column Manager's In-Line Filter Unit
Swing the column retainer rod out to the right-hand side. Column retainer rod Using the Phillips screwdriver, loosen the 2 screws that fasten the column stabilizer assembly to the tray. Remove the column stabilizer assembly from the thermal gasket. Using the 1/4-inch and 5/16-inch open-end wrenches, disconnect the compression fitting that attaches the column stabilizer assembly to the optional in-line filter or to the column.
Page 373 To replace the filter in the column in-line filter unit Stop any currently running mobile phase before replacing the column in-line filter unit. Open the column manager door and locate the column in-line filter unit you want to replace. Column number 1 is at the top position, and column number 4 is at the bottom.
Page 374 Remove the column components (column, in-line filter unit, and stabilizer tubing assembly). Flow In-line filter unit From column stabilizer Column tubing assembly Inlet TP02873 Reusable fitting with holes for spiked end of collet tool Using the 5/16-inch open-end wrench to hold the in-line filter unit, use the spiked end of the collet separator to turn the reusable fitting.
Page 375: Replacing The Filter In The Column Heater/Cooler's In-Line Filter Unit
Insert a new frit, and then reassemble the column in-line filter unit using the wrenches. 10. Reattach the column stabilizer tubing assembly to the in-line filter unit. 11. Start the mobile phase and examine the assembled components for leaks. Tighten fittings as needed. 12.
Page 376 Swing the column retainer rod out to the right-hand side. Column retainer rod Remove the column components (column, in-line filter unit, and stabilizer tubing assembly). Flow In-line filter unit From column stabilizer Column tubing assembly Inlet TP02873 Reusable fitting with holes for spiked end of collet tool Older reusable fittings do not have holes in the side of them.
Page 377: Cleaning The Column Manager And Column Heater/Cooler
Over time, the filter can expand inside the in-line filter inlet nut. If Tip: you are unable to remove the filter from the inlet nut, discard the inlet nut and replace both the inlet nut and filter. Insert a new filter, and then reassemble the column in-line filter unit using the wrenches.
Page 378: Interior Surfaces
You can remove the drainage cup and flush it out in the sink (this waste is not hazardous). If an ACQUITY UPLC detector (TUV or PDA) is installed on top of the column manager or column heater/cooler, leaks and drainage from the detectors merge with the leak path of the column manager or column heater/cooler.
Page 379: Maintaining The Tuv Detector
To avoid damaging the column, remove it before you flush the Caution: system. If your system includes a PDA detector, see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide for information on maintaining it. If your system includes an ELS detector, see the ACQUITY UPLC Evaporative Light Scattering Detector Getting Started Guide for information on maintaining it.
Page 380: Resolving Detector Leak Sensor Errors
Nonabrasive, lint-free wipes To resolve a detector leak sensor error View the Leak Sensors dialog box in the ACQUITY UPLC Console to confirm that the detector leak sensor detected a leak. If a leak is detected, a “Leak Detected” error message appears.
Page 381 Remove the leak sensor from its reservoir by grasping it by its serrations and pulling upward on it. Serrations If you cannot easily manipulate the leak sensor after removing it Tip: from its reservoir, detach the connector from the front of the instrument (see page 6-208).
Page 382 Roll up a nonabrasive, lint-free wipe, and use it to absorb the liquid from the leak sensor reservoir and its surrounding area. Rolled up lint-free wipe Leak sensor reservoir With a cotton swab, absorb any remaining liquid from the corners of the leak sensor reservoir and its surrounding area.
Page 383 If you detached the connector from the front of the instrument, reattach 10. In the ACQUITY UPLC Console, select your detector from the system tree. 11. In the detector information window, click Control > Reset, to reset the detector.
Page 384: Replacing The Detector's Leak Sensor
Replacing the detector’s leak sensor The leak sensor can be contaminated with biohazardous Warning: and/or toxic materials. Always wear clean, chemical-resistant, powder-free gloves when performing this procedure. Required materials • Clean, chemical-resistant, powder-free gloves • Leak sensor To replace the detector leak sensor Open the detector door, gently pulling its right-hand edge toward you.
Page 385 Remove the leak sensor from its reservoir by grasping it by its serrations and pulling upward on it. Serrations Unpack the new leak sensor. Maintaining the TUV detector 6-209...
Page 386: Maintaining The Flow Cell
Leak sensor installed in reservoir Plug the leak sensor connector into the front of the instrument. In the ACQUITY UPLC Console, select your detector from the system tree. In the detector information window, click Control > Reset, to reset the detector.
Page 387 refractive index of the fluid is greater than that of the tubing material (Teflon AF). Light transmission through light guiding flow cell Light path Teflon AF α Mobile phase Teflon AF Teflon AF In the figure, above, the light path through the cell is depicted by a pair of rays (dashed lines) that bounce off the cell wall.
Page 388: Cleaning The Flow Cell
The light path through a conventional flow cell is designed to avoid contact with the cell walls, primarily to prevent rays that encounter the walls from contributing to the measured signal. The energy associated with errant rays is highly variable depending on the mobile phase composition, quality of wall finish (which is never 100% reflective) or the slow build-up of contaminants.
Page 389 “Performing a system acid cleansing flush” on page 6-215). • If there is still no improvement, call Waters Technical Service. Caution: • To prevent flow cell failure, do not connect any tubing or device that can create backpressure exceeding the flow cell’s maximum rating of 6895 kPa (69 bar, 1000 psi).
Page 390 To clean the flow cell In the detector control panel, click (Lamp Off). Stop the solvent flow and remove the column. Replace the column with a union or piece of tubing. If another instrument is downstream of the flow cell outlet, break the connection at the other instrument and route the outlet tubing to waste while flushing.
Page 391: Performing A System Acid Cleansing Flush
If you are running an MS detector, do not perform the Caution: system acid cleansing flush. Instead, call Waters Technical Service. To prepare the solvent Prepare a mixture of 50:50 (v/v) methanol:water as follows: Measure 500 mL of water in a graduated cylinder.
Page 392 Place all lines A1, A2, B1, B2 seal wash, weak needle wash and strong needles in 50:50 methanol:water. Prime the solvent lines for 5 minutes each. Prime the seal wash. Prime the wash syringes and sample syringe for 4 cycles. Connect a pressure restrictor in the fluid path after the injector, to create 13,790 kPa (138 bar, 2000 psi) backpressure in the system.
Page 393: Replacing The Flow Cell
• To avoid damaging the flow cell, handle it with care. Do not disassemble the flow cell. Controlling Contamination in Ultra Performance LC/MS and See also: HPLC/MS Systems (part number 715001307) on the ACQUITY UPLC System Bookshelf CD. Required materials •...
Page 394 Disconnect the detector’s inlet and outlet tubing from the main column connection. Outlet tubing Flow cell assembly Captive screws Inlet tubing TP02952 6-218 Maintaining the System...
Page 395 Remove the flow cell: • Using a 1/4-inch, flat-blade screwdriver, loosen the 3 captive screws on the flow cell assembly’s front plate. • Grasp the handle and gently pull the assembly toward you. Captive screw Handle Captive screws TP02952 To avoid damaging the capillary tubing, do not touch it. Caution: Unpack and inspect the new flow cell.
Page 396 Top view of the analytical flow cell assembly Flow cell body Capillary tubing Front of flow cell assembly TP02583 Top view of the high sensitivity flow cell assembly Flow cell body Capillary tubing Front of flow cell assembly TP02582 Insert the new flow cell assembly into the detector, and then tighten the captive screws.
Page 397: Replacing The Lamp
Record the new lamp’s serial number to properly track lamp life. If you do Tip: not record a new lamp’s serial number in the ACQUITY UPLC Console, the date of the previous lamp installation remains in the detector’s memory. To prevent burn injuries, allow the lamp to cool for 30 Warning: minutes before removing it.
Page 398 Power-off the detector and disconnect the power cable from the rear panel. The lamp and lamp housing can be hot. Wait 30 Warning: minutes after powering off the detector for these components to cool before touching them. Allow the lamp to cool for 30 minutes, and then open the door, gently pulling its right-hand edge toward you.
Page 399: Replacing The Fuses
If your detector has the power entry module shown below and it fails to power-on, the status LEDs are unlit, or the power supply fan does not operate, call Waters Technical Service (see page 6-6).
Page 400 To avoid electric shock, power-off and unplug the TUV Warning: detector before examining the fuses. For continued protection against fire, replace fuses with those of the same type and rating only. Required material 100 to 240 VAC, 50 to 60-Hz, F 3.15-A, 250-V (fast-blow), 5 × 20 mm (IEC) fuses (2) Suspect a fuse is open or otherwise defective when •...
Page 401: Cleaning The Instrument's Exterior
Remove and discard the fuses. Make sure that the new fuses are properly rated for your requirements, and then insert them into the holder and the holder into the power entry module, gently pushing until the assembly locks into position. Reconnect the power cord to the power entry module.
Page 402: Replacing The Fuses
If your sample organizer has the power entry module shown below and it fails to power-on, the status LEDs are unlit, or the power supply fan does not operate, call Waters Technical Service (see page 6-6).
Page 403 To avoid electric shock, power-off and unplug the sample Warning: organizer before examining the fuses. For continued protection against fire, replace fuses with those of the same type and rating only. Suspect a fuse is open or otherwise defective when •...
Page 404: Cleaning The Instrument's Exterior
With minimum pressure, pull on each spring-loaded fuse holder to remove it. Fuse Fuse holder door Fuse holder Power entry module Remove and discard the fuses. For continued protection against fire, replace fuses Warning: with the appropriate type and rating. Make sure that the new fuses are properly rated for your requirements, and then insert the new fuses into the holders and the holders into the power entry module.
Page 405: Maintaining The 30-Cm Column Heater/Cooler
If your 30-cm column heater/cooler has the power entry module shown below and it fails to power-on, the status LEDs are unlit, or the power supply fan does not operate, call Waters Technical Service (see page 6-6).
Page 406: Cleaning The Instrument's Exterior
To replace the fuses Replace both fuses, even when only one is open or defective. Requirement: To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to an instrument. To completely interrupt power to an instrument, set the power switch to Off, and then unplug the power cord from the AC outlet.
Page 407: Troubleshooting
Troubleshooting This chapter describes how to find the cause of problems that can affect ® performance of the ACQUITY UPLC system and how to correct them. Contents Topic Page Troubleshooting with Connections Insight Proper operating procedures Solving baseline noise and drift problems...
Page 408: Troubleshooting With Connections Insight
The service agent software automatically and securely captures and sends information about the support needs of your system directly to Waters. If you encounter a performance issue as you are using the ACQUITY UPLC Console, you can also manually submit a Connections Insight request to Waters customer support.
Page 409: Proper Operating Procedures
A .ZIP file containing your Connections Insight profile is Result: forwarded to Waters customer support for review. Saving a service profile or plot file from the ACQUITY UPLC Console can require up to 150 MB of file space. Proper operating procedures So that your system always runs optimally, follow the operating procedures and guidelines in Chapters 2 through 6.
Page 410: Locating System Serial Numbers
Be prepared to provide the serial numbers of the Recommendation: instruments in your system when you contact Waters customer support. To view instrument information In the ACQUITY UPLC Console, select an instrument from the system tree. Troubleshooting...
Page 411: Solving Baseline Noise And Drift Problems
Click Configure > View module information. The Module Information dialog box displays this information: • Serial number • Firmware version • Firmware checksum • Component software version Alternatives: • From the main window, place the pointer over the visual representation of the system instrument you want information for.
Page 412 Corrective actions for baseline noise and drift problems (Continued) Symptom Possible cause Corrective action Flat baseline with no No solvent flow 1. Check the solvent peaks reservoirs and ensure they contain solvent. 2. Make sure the flow rate is > 0. Detector lamp not •...
Page 413 Injector not injecting 1. Prime the sample syringes (see page 3-17). 2. Run the sample syringe leak test (see the ACQUITY UPLC Console online Help). 3. If the problem persists, call Waters Technical Service (see page 7-3). Method parameters Set correct method incorrect parameters.
Page 414 Corrective actions for baseline noise and drift problems (Continued) Symptom Possible cause Corrective action Random baseline Air in detector flow 1. Flush the detector flow cell noise cell to remove air. 2. Check the backpressure regulator and ensure it is properly installed.
Page 415 Corrective actions for baseline noise and drift problems (Continued) Symptom Possible cause Corrective action Random baseline Low lamp energy 1. Check the lamp energy (see noise (continued) the ACQUITY UPLC Console online Help). 2. Replace the lamp, if necessary (see page 6-221). Rapid baseline shift Column not Equilibrate the column.
Page 416: Binary Solvent Manager Troubleshooting
UV detector flow cell Inspect flow cell, tighten leaking connections. If flow cell is leaking, call Waters Technical Service (see page 7-3). Dirty flow cell Clean the flow cell. There is an Balance the absorbance of the...
Page 417: Flow Led
Flashing red Indicates a serious error that has stopped pump operation. Information about the error can be found in the ACQUITY UPLC Console. Steady red Indicates a serious failure within the binary solvent manager that prevents further operation.
Page 418 1. Prime binary solvent pressures (continued) manager (see page 3-9). 2. Perform the static decay test to identify faulty check valve (see ACQUITY UPLC Console online Help). 3. Replace faulty check valve (see page 6-46). High-pressure leak in Locate and repair the...
Page 419 Fans do not run Power supply not 1. Reset the binary (continued) working solvent manager via the control panel or ACQUITY UPLC Console. 2. If the problem persists, power-off and power-on again. 3. If the problem persists, call Waters Technical Service (see page 7-3).
Page 420: Sample Manager Troubleshooting
1. Reset the binary failure controller board, solvent manager via solvent management the control panel or system, or sample ACQUITY UPLC management system Console. 2. If the problem persists, power-off and power-on again. 3. If the problem persists, call Waters...
Page 421 Flashing red Indicates a serious error that prevents further operation. You can find information about the error in the ACQUITY UPLC Console. Steady red Indicates a serious failure within the sample manager that prevents further operation. Call Waters Technical Service (see page 7-3).
Page 422 ACQUITY UPLC Console. 2. If the problem persists, power-off and power-on again. 3. If the problem persists, call Waters Technical Service (see page 7-3). Power supply not 1. Reset the sample working manager via the...
Page 423: Sample Organizer Troubleshooting
Sample manager troubleshooting (Continued) Symptom Possible cause Corrective action Leak detected Leak in sample Follow the procedure to manager. resolve a sample manager leak sensor error on page 6-123. Sample compartment Sample compartment 1. Defrost sample freezes door opened too manager, and then frequently choose a sample...
Page 424 Flashing red Indicates a serious error that prevents further operation. You can find information about the error in the ACQUITY UPLC Console. Steady red Indicates a serious failure within the sample organizer that prevents further operation. Call...
Page 425 Power supply fan 1. Reset the sample (continued) wiring or motor organizer via the problem sample manager control panel or ACQUITY UPLC Console. 2. If the problem persists, power-off and power-on again. 3. If the problem persists, call Waters Technical Service (see page 7-3).
Page 426 Corrective action Sample plates jam Nonconforming vial or Ensure that all plates microtiter plates conform to ANSI standards. Bad plate sensor fiber Call Waters Technical optic Service (see page 7-3). Incorrect alignment of Call Waters Technical X and Y stops...
Page 427: Ht Column Heater Troubleshooting
HT column heater troubleshooting HT column heater troubleshooting Symptom Possible cause Corrective action Leak detected Leak in HT column Follow the procedure to heater. resolve a HT column heater leak sensor error page 6-162 page 6-167. Column manager and column heater/cooler troubleshooting Power LED The power LED, on the left-hand side of the sample manager’s front panel,...
Page 428 Flashing red Indicates a serious error that prevents further operation. You can find information about the error in the ACQUITY UPLC Console. Steady red Indicates a serious failure within the column manager or column heater/cooler that prevents further operation. Call Waters Technical Service...
Page 429 Power supply fan 1. Reset the column wiring or motor manager or column problem heater/cooler via control panel or ACQUITY UPLC Console. 2. If the problem persists, power-off and power-on again. 3. If the problem persists, call Waters Technical Service...
Page 430: Tuv Detector Troubleshooting
Steady red color Indicates a serious error that prevents further operation. You can find information about the error in the ACQUITY UPLC Console. Flashing red color Indicates a serious failure within the detector that prevents further operation. Call Waters...
Page 431 Establish liquid flow through the detector flow cell with degassed acetonitrile or methanol at a flow rate that is anticipated to be used for subsequent analysis. TUV detector troubleshooting Symptom Possible cause Corrective action Analog output incorrect AUFS setting changed Reset the AUFS setting.
Page 432 (see page 6-221). Lamp input shorted in Inspect the rear panel “off” position connections (see ACQUITY UPLC Console online Help) or look for a timed event within a method (possibly programmed to extinguish the lamp). Bad lamp power supply 1.
Page 433: Pda Detector Troubleshooting
TUV detector troubleshooting (Continued) Symptom Possible cause Corrective action No sample and Lamp burned out Attempt to reignite the reference energy; lamp lamp. If it fails to LED is off, solid red, or ignite, replace the flashing red lamp. Lamp extinguished 1.
Page 434: Lamp Led
Steady red color Indicates a serious error that prevents further operation. You can find information about the error in the ACQUITY UPLC Console. Flashing red color Indicates a serious failure within the detector that prevents further operation. Call Waters...
Page 435 Air bubbles trapped in • Reseat and check flow cell alignment of flow cell (see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide). • Flush the flow cell (see the ACQUITY UPLC Photodiode Array Detector...
Page 436 PDA detector troubleshooting (Continued) Symptom Possible cause Corrective action Detector not Bad or disconnected Inspect cable responding to cable connections, tighten ACQUITY UPLC connectors, or replace Console cable. Configuration problem Check Ethernet configuration. For details, see Empower online Help. Leak detected...
Page 437 (see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide). Dirty flow cell causing Flush the flow cell (see shutter diagnostic test the ACQUITY UPLC to fail Photodiode Array Detector Getting Started Guide). Insufficient energy To prevent air bubbles reaching photodiode...
Page 438: Els Detector Troubleshooting
Steady red color Indicates a serious error that prevents further operation. You can find information about the error in the ACQUITY UPLC Console. Flashing red color Indicates a serious failure within the detector that prevents further operation. Call Waters...
Page 439: Flr Detector Troubleshooting
2. Test electrical outlet for power. Cable loose or defective Inspect line cord connections. Open (spent) or Replace fuse (see the defective fuse ACQUITY UPLC Evaporative Light Scattering Detector Getting Started Guide). Communication Configuration problem Check Ethernet problems configuration.
Page 440: Lamp Led
Constant red Indicates a detector failure that prevents further operation. Power-off the detector, and then power-on. If the LED is still steady red, contact your Waters service representative. FLR detector troubleshooting Symptom Possible cause Corrective action Both LEDs unlit No power 1.
Page 441: Chromatography Troubleshooting
UPLC system. Know the physical and chemical properties of the solvents you use. See the Material Safety Data Sheets for the solvents in use. The table below lists chromatographic symptoms, possible causes, and suggested corrective actions for the ACQUITY UPLC system. Chromatography troubleshooting 7-35...
Page 442 For a 2.1 × 100 mm Example: ACQUITY UPLC BEH column, run about 2.5 mL of solution at 0.5 mL/min for about 5 minutes. ACQUITY UPLC BEH See also: Column Care and Use Instructions.
Page 443 2. If symptom persists, clean the column using the cleaning and regeneration procedures described in Waters ACQUITY UPLC BEH Column Care and Use Instructions. 3. If system persists, replace the column. Decreased Incorrect flow Change flow rate.
Page 444 Chromatography troubleshooting (Continued) Symptom Possible cause Corrective action Erratic retention Air bubble in Prime binary solvent manager. times pump head. Malfunctioning Clean or replace inlet check valve check valves. cartridges. Leaking plunger Replace seals (see page 6-52). seals. Separation Determine the suitability of mobile chemistry.
Page 445 2. If symptom persists, clean the column using the cleaning and regeneration procedures described in Waters ACQUITY UPLC BEH Column Care and Use Instructions. 3. If symptom persists, replace the column. Tubing is Systematically check tubing by plugged.
Page 446 For a 2.1 × 100 mm Example: ACQUITY UPLC BEH column, run about 2.5 mL of solution at 0.5 mL/min for about 5 minutes. ACQUITY UPLC BEH See also: Column Care and Use Instructions.
Page 447 Chromatography troubleshooting (Continued) Symptom Possible cause Corrective action Reproducibility Sample Troubleshoot sample management errors (continued) management system. system problem. Incorrect wash Change wash solvents and/or solvents or volumes. volumes. Incorrect volume Change volume range. range for injection method or type. Sensitivity loss Leak in binary Troubleshoot solvent management...
Page 448 7-42 Troubleshooting...
Page 449: A Safety Advisories
Safety Advisories Waters instruments display hazard symbols designed to alert you to the hidden dangers of operating and maintaining the instruments. Their corresponding user guides also include the hazard symbols, with accompanying text statements describing the hazards and telling you how to avoid them.
Page 450: Warning Symbols
Heed all warnings when you install, repair, and operate Waters instruments. Waters assumes no liability for the failure of those who install, repair, or operate its instruments to comply with any safety precaution.
Page 451: Warnings That Apply To Particular Instruments, Instrument Components, And Sample Types
The following warnings can appear in the user manuals of particular instruments and on labels affixed to them or their component parts. Burst warning This warning applies to Waters instruments fitted with nonmetallic tubing. Pressurized nonmetallic, or polymer, tubing can burst. Warning: Observe these precautions when working around such tubing: •...
Page 452: Biohazard Warning
Standby mode before touching areas marked with this high voltage warning symbol. Biohazard warning This warning applies to Waters instruments that can be used to process material that might contain biohazards: substances that contain biological agents capable of producing harmful effects in humans.
Page 453: Caution Symbol
Chemical hazard warning This warning applies to Waters instruments that can process corrosive, toxic, flammable, or other types of hazardous material. Waters instruments can be used to analyze or Warning: process potentially hazardous substances. To avoid injury with any of these materials, familiarize yourself with the materials and their hazards, observe Good Laboratory Practices (GLP), and consult your organization’s safety...
Page 454 Attention: Changes or modifications to this unit not expressly approved by the party responsible for compliance could void the user’s authority to operate the equipment. Important: Toute modification sur cette unité n’ayant pas été expressément approuvée par l’autorité responsable de la conformité à la réglementation peut annuler le droit de l’utilisateur à...
Page 455 • Keine Schläuche verwenden, die stark geknickt oder überbeansprucht sind. • Nichtmetallische Schläuche nicht für Tetrahydrofuran (THF) oder konzentrierte Salpeter- oder Schwefelsäure verwenden. • Durch Methylenchlorid und Dimethylsulfoxid können nichtmetallische Schläuche quellen; dadurch wird der Berstdruck des Schlauches erheblich reduziert. Warnings that apply to all Waters instruments...
Page 456 Attenzione: fare attenzione quando si utilizzano tubi in materiale polimerico sotto pressione: • Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero pressurizzati. • Spegnere tutte le fiamme vive nell'ambiente circostante. • Non utilizzare tubi eccessivamente logorati o piegati. •...
Page 457 농축 질산 또는 황산과 함께 사용하지 마십시오. 염화 메틸렌(Methylene chloride) 및 디메틸술폭시드(Dimethyl sulfoxide)는 비금속 튜브를 부풀려 튜브의 파열 압력을 크게 감소시킬 수 있으므로 유의하십시오. 警告：圧力のかかったポリマーチューブを扱うときは、注意してください。 • 加圧されたポリマーチューブの付近では、必ず保護メガネを着用してください。 • 近くにある火を消してください。 • 著しく変形した、または折れ曲がったチューブは使用しないでください。 • 非金属チューブには、テトラヒドロフラン(THF)や高濃度の硝酸または硫酸などを流さないでください。塩化メチレンやジメチルスルホキシドは、非金属チューブの膨張を引き起こす場合があり、その場合、チューブは極めて低い圧力で破裂します。 Warnings that apply to all Waters instruments...
Page 458 Warning: The user shall be made aware that if the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired. Attention: L’utilisateur doit être informé que si le matériel est utilisé d’une façon non spécifiée par le fabricant, la protection assurée par le matériel risque d’être défectueuses.
Page 459 警告 : 為了避免火災，更換保險絲時，請使用與儀器保險絲蓋旁面板上所印刷之相同類型與規格的保險絲。警告 : 为了避免火灾，应更换与仪器保险丝盖旁边面板上印刷的类型和规格相同的保险丝。 경고: 화재의 위험을 막으려면 기기 퓨즈 커버에 가까운 패널에 인쇄된 것과 동일한 타입 및 정격의 제품으로 퓨즈를 교체하십시오. 警告 : 火災予防のために、ヒューズ交換では機器ヒューズカバー脇のパネルに記載されているタイプおよび定格のヒューズをご使用ください。 Warnings that apply to all Waters instruments A-11...
Page 460: Electrical And Handling Symbols
Electrical and handling symbols Electrical symbols These can appear in instrument user manuals and on the instrument’s front or rear panels. Electrical power on Electrical power off Standby Direct current Alternating current Protective conductor terminal Frame, or chassis, terminal Fuse Recycle symbol: Do not dispose in municipal waste.
Page 461: Handling Symbols
Handling symbols These handling symbols and their associated text can appear on labels affixed to the outer packaging of Waters instrument and component shipments. Keep upright! Keep dry! Fragile! Use no hooks! Electrical and handling symbols A-13...
Page 462 A-14 Safety Advisories...
Page 463: B Specifications
Specifications The system specifications outlined in this appendix depend on the conditions in your laboratory. Refer to the ACQUITY UPLC Site Preparation Guide or contact Waters Technical Service for more information on specifications. Contents: Topic Page Binary solvent manager specifications...
Page 464: Binary Solvent Manager Specifications
Binary solvent manager specifications Binary solvent manager environmental specifications Attribute Specification Operating temperature 4 to 40 °C (39.2 to 104 °F) Operating humidity <50%, noncondensing Shipping and storage temperature −30 to 60 °C (-22 to 140 °F) <90%, noncondensing Shipping and storage humidity Binary solvent manager electrical specifications Attribute Specification...
Page 465 Binary solvent manager performance specifications Attribute Specification Number of solvents Up to four, in any combination of two: A1 or A2 and B1 or B2. Solvent storage Solvent tray accommodates up to four chromatographic solvents, two sample manager wash solvents, and one binary solvent manager seal wash solvent.
Page 466: Sample Manager Specifications
PEEK alloys, titanium alloys. Unattended operation Leak sensors, full 96-hour diagnostic data displayed through ACQUITY UPLC console software a. Contact Waters for the conditions used to derive this attribute. Sample manager specifications Sample manager environmental specifications Attribute Specification 4 to 40 °C (39.2 to 104 °F)
Page 467 8 deep-well (or 2 mL vial) plates: 720002449EN) • 96 and 384 microtiter plates • Sample Vials and Accessories for • 48-position 2 mL vial plates all ACQUITY UPLC, HPLC, and GC Systems (part number • 48-position 0.65 mL 720001818EN) micro-centrifuge tube plates •...
Page 468 Injector linearity >0.999 coefficient of deviation (from 20 to 75% of the loop volume, partial loop overfill mode (PLNO injection mode) per Waters ACQUITY UPLC AQT/SystemsQT protocol Sample temperature control 4 to 40 °C, programmable in 0.1 °C increments [assumes ambient temperature of 21 °C (69.8 °F)]...
Page 469: Sample Organizer Specifications
Sample manager performance specifications (Continued) Attribute Specification Minimum sample required 5 µL residual volume at a needle offset of 0, or 22 µL at the preset 2 mm needle offset Wash solvents Two degassed solvents: strong solvent and weak solvent, programmable wash volumes to suit application Sample carryover <0.005% or <2.0 nL, whichever is...
Page 470 Sample organizer electrical specifications Attribute Specification Class I Protection class Overvoltage category Pollution degree Normal (IPXO) Moisture protection Grounded AC Line voltages, nominal a. Protection Class I – The insulating scheme used in the instrument to protect from elec- trical shock. Class I identifies a single level of insulation between live parts (wires) and exposed conductive parts (metal panels), in which the exposed conductive parts are con- nected to a grounding system.
Page 471 • Waters Vials for ACQUITY combinations thereof. UPLC Systems (part number 720002449EN) • Sample Vials and Accessories for all ACQUITY UPLC, HPLC, and GC Systems (part number 720001818EN) Maximum sample capacity Total of up to 8,064 samples: in twenty one 384-well plates.
Page 472 Sample organizer thermal specification Maximum compartment temperature +6 °C Compartment setpoint temperature −2 °C Minimum compartment temperature Ambient temperature (°C) B-10 Specifications...
Page 473: Ht Column Heater Specifications
HT column heater specifications HT column heater environmental specifications Attribute Specification Operating temperature 4 to 40 °C (39.2 to 104 °F) Operating humidity <50%, noncondensing Shipping and storage temperature −30 to 60 °C (-22 to 140 °F) <90%, noncondensing Shipping and storage humidity HT column heater electrical specifications Attribute Specification...
Page 474 Over temperature Thermal switch shuts (switch opens) off heater at temperatures greater than 115 °C ±4.4 °C. Switch recloses at 100 °C ±3.3 °C Unattended operation Leak sensors, full 96-hour diagnostic data displayed through ACQUITY UPLC console software B-12 Specifications...
Page 475: Column Manager And Column Heater/Cooler Specifications
Column manager and column heater/cooler specifications Column manager and column heater/cooler environmental specifications Attribute Specification Operating temperature 4 to 40 °C (39.2 to 104 °F) Operating humidity <50%, noncondensing −30 to 60 °C (-22 to 140 °F) Shipping and storage temperature <90%, noncondensing Shipping and storage humidity Column manager and column heater/cooler electrical specifications...
Page 476 Thermal switch shuts (switch opens) off heater at temperatures greater than 115 °C ±4.4 °C. Switch recloses at 100 °C ±3.3 °C. Unattended operation Leak sensors, full 96-hour diagnostic data displayed through ACQUITY UPLC console software a. Column manager only. B-14 Specifications...
Page 477: 30-Cm Column Heater/Cooler Specifications
30-cm column heater/cooler specifications Column manager environmental specifications Attribute Specification Operating temperature 4 to 40 °C (39.2 to 104 °F) Operating humidity <50%, noncondensing Shipping and storage temperature −30 to 60 °C (-22 to 140 °F) <90%, noncondensing Shipping and storage humidity Column manager electrical specifications Attribute Specification...
Page 478 Temperature measured by an NIST traceable probe located next to the measurement sensor will be within the specification, per Waters ACQUITY UPLC AQT/SystemsQT protocol. Over temperature Thermal switch shuts off heater at temperatures greater than 80.0 °C (176 °F) ±10% (temperature measured at thermal...
Page 479: Tuv Detector Specifications
TUV detector specifications TUV detector environmental specifications Attribute Specification Operating temperature 4 to 40 °C (39.2 to 104 °F) Operating humidity <50%, noncondensing Shipping and storage temperature −30 to 60 °C (-22 to 140 °F) <90%, noncondensing Shipping and storage humidity TUV detector electrical specifications Attribute Specification...
Page 480 0.0125 to 5.0 seconds, Hamming (sampling rate dependent) Dual wavelength: 0.5 to 5.0 seconds, Hamming (sampling rate dependent) Unattended operation Leak sensors, full 96-hour diagnostic data displayed through ACQUITY UPLC console software Optical Component Specifications Lamp source Deuterium arc lamp Lamp warranty 2000 hours...
Page 481 2400 69/1000 sensitivity If your system includes a PDA detector, see the ACQUITY UPLC Photodiode Array Detector Getting Started Guide for specifications. If your system includes an ELS detector, see the ACQUITY UPLC Evaporative Light Scattering Detector Getting Started Guide for specifications.
Page 482 B-20 Specifications...
Page 483: C Solvent Considerations
The information in this appendix applies only to the following instruments: ® • ACQUITY UPLC system modules • ACQUITY UPLC PDA detector • ACQUITY UPLC PDA eλ detector • ACQUITY UPLC TUV detector • ACQUITY UPLC ELS detector • ACQUITY UPLC FLR detector Contents...
Page 484: Introduction
Preventing contamination For information on preventing contamination, refer to Controlling Contamination in Ultra Performance LC/MS and HPLC/MS Systems (part number 715001307) on the ACQUITY UPLC System Bookshelf CD, or visit http://www.waters.com and click Waters Division > Services & Support. Clean solvents Clean solvents ensure reproducible results and permit you to operate with minimal instrument maintenance.
Page 485: Water
For information on preventing contamination, refer to Controlling See also: Contamination in Ultra Performance LC/MS and HPLC/MS Systems (part number 715001307) on the ACQUITY UPLC System Bookshelf CD, or visit http://www.waters.com and click Waters Division > Services & Support. Introduction...
Page 486: Solvent Recommendations
ACQUITY UPLC BEH column chemistries. Waters evaluated the system’s reliability using traditional reversed-phase solvents. This section lists solvents recommended for the ACQUITY UPLC system. Contact Waters Customer Service to determine whether you can use solvents that do not appear in the list without adversely affecting instrument or system performance.
Page 487 Hexane/THF Compatibility Kit The ACQUITY UPLC System Hexane/THF Compatibility Kit (part number 205000464) can be installed in ACQUITY UPLC systems with closed waste management. It is designed for users that need to run their systems 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.
Page 488: Solvents To Avoid
Cleaning agents See the cleaning procedures in Controlling Contamination Recommendation: in Ultra Performance LC/MS and HPLC/MS Systems (part number 715001307) on the ACQUITY UPLC System Bookshelf CD, or visit http://www.waters.com and Waters Division > Services & Support. • Phosphoric acid (≤30%) •...
Page 489 • THF and hexane can be used as the mobile-phase in ACQUITY UPLC systems. However, as with many nonaqueous solvents, they can shorten system and instrument life compared with equipment running typical reversed-phase solvents.
Page 490: Binary Solvent Manager Recommendations
Binary solvent manager recommendations • The seal wash system must never run dry, particularly during separations that use a polar mobile phase. • Isopropyl alcohol or mixtures of methanol and water, like 20% methanol/water, are effective seal wash solvents for THF solvent mixtures.
Page 491: Using Miscibility Numbers (M-Numbers
When you switch from a strong buffer to an organic solvent, thoroughly flush the system with distilled water before you add the organic solvent. Solvent miscibility Viscosity Boiling Miscibility λ Cutoff Polarity Solvent cP, 20 °C point °C number index (nm) (@1 atm) (@1 atm)
Page 492: Solvent Stabilizers
Some solvents prove immiscible with solvents at both ends of the lipophilicity scale. These solvents receive a dual M-number: • The first number, always lower than 16, indicates the degree of miscibility with highly lipophilic solvents. • The second number applies to the opposite end of the scale. A large difference between these two numbers indicates a limited range of miscibility.
Page 493: Wavelength Selection
Wavelength selection The tables in this section provide UV cutoff values for these items: • Common solvents • Common mixed mobile phases • Chromophores UV cutoffs for common solvents The table below shows the UV cutoff (the wavelength at which the absorbance of the solvent equals 1 AU) for some common chromatographic solvents.
Page 494 10 mM, pH 7.6 Hydrochloric acid, 0.1% ® Waters PIC Reagent A, 1 vial/liter Morpholinoethanesulfonic acid Waters PIC Reagent B-6, (MES), 10 mM, pH 6.0 1 vial/liter Potassium phosphate, Waters PIC Reagent B-6, monobasic, 10 mM low UV, 1 vial/liter...
Page 495 The best mobile phase for your application is one that is transparent at the chosen detection wavelengths. With such a mobile phase, ensure that any absorbance is due only to the sample. Absorbance by the mobile phase also reduces the linear dynamic range of the detector by the amount of absorbance the autozero function cancels, or “autozeroes”, out.
Page 496 Mobile phase absorbance measured against air or water (Continued) Absorbance at specified wavelength (nm) Trifluoro- 1.20 0.78 0.54 0.34 0.22 0.06 <0.02 <0.01 — — acetic acid Diammo- 1.85 0.67 0.15 0.02 <0.01 — — — — — nium phosphate, 50 mM Triethy- 2.33...
Page 497 Mobile phase absorbance measured against air or water (Continued) Absorbance at specified wavelength (nm) Morpholino- 2.42 2.38 1.89 0.90 0.45 0.06 <0.01 — — — ethanesulfo nic acid (MES), 10 mM, pH 6.0 Potassium 0.03 <0.01 — — — — —...
Page 498 — 20 mM, pH Tris HCl, 1.80 1.90 1.11 0.43 0.13 <0.01 — — — — 20 mM, pH ® Waters PIC reagents 0.67 0.29 0.13 0.05 0.03 0.02 0.02 0.02 0.02 <0.01 PIC A, 1 vial/L PIC B6, 1 2.46...
Page 499 Mobile phase absorbance measured against air or water (Continued) Absorbance at specified wavelength (nm) Detergents BRI J 35, 1% 0.06 0.03 0.02 0.02 0.02 0.01 <0.01 — — — 3-[(3-chola- 2.40 2.32 1.48 0.80 0.40 0.08 0.04 0.02 0.02 0.01 midopropyl) -dimethy- lammonio]-1...
Page 500 C-18 Solvent Considerations...
Page 501 6-46 reattaching 6-116 accumulator head acids C-13 reattaching 6-116 acquisition speed 1-11 removing 6-103 ACQUITY UPLC system accumulator head plunger, columns calculator replacing 6-114 Console air filters starting from Empower cleaning 6-51 software 4-12 replacing 6-52...
Page 502 I/O signal connectors 2-59 vent valve, replacing 6-118 installing 2-10 caution symbol maintaining characterizing overview needle seal 3-21 3-22 6-140 plumbing 2-26 needle volume 3-22 6-141 power LED 7-10 7-17 sample loop volume 3-22 6-141 preparing chart-out parameters 2-61 primary head check valve, replacing 6-19 reattaching...
Page 503 2-57 specifications signal, making 2-65 electrical B-13 Connections Insight request, environmental B-13 submitting performance B-14 contacting Waters Technical Service troubleshooting 7-21 7-23 – see eCord column chip contamination, preventing column manager control panel control panel, using 4-11 binary solvent manager...
Page 504 6-212 overview 1-12 drain routing hole 2-14 starting draw rates, maximum 6-149 6-150 starting ACQUITY UPLC drift problems, troubleshooting Console from 4-12 – 7-10 system test mix, preparing dry prime, performing 3-10 system test mix running...
Page 505 FLR detector control panel, using ferrule installation 6-148 I/O signal connectors 2-64 filter lamp LED 7-34 binary solvent manager maintaining 6-203 cleaning 6-51 overview 1-11 replacing 6-52 plumbing 2-25 column in-line, replacing 6-178 power LED 7-33 6-197 6-199 preparing 3-36 sample organizer, cleaning 6-225 signal connectors...
Page 506 overview sample manager 2-10 set point sample organizer specifications shade, sample manager 3-28 electrical B-11 instrument environmental B-11 cleaning performance B-12 information, viewing troubleshooting 7-21 method, creating see eCord column chip selecting intended use ISM classification I/O signal connectors binary solvent manager 2-59 ELS detector 2-64...
Page 507 4-13 lamp 4-10 7-24 overview 1-12 power 7-24 starting 4-13 lights, turning off starting ACQUITY UPLC Console load ahead mode overview 3-26 from 4-14 loading sample plates 3-24 3-31 median baseline filter 1-11 logging in miscibility Empower software...
Page 508 troubleshooting 7-27 7-31 – verifying operational specifications, TUV performance specifications detector B-18 binary solvent manager organizer, see sample organizer column 30-cm column heater/cooler overview B-16 30-cm column heater/cooler column heater/cooler B-14 binary solvent manager column manager B-14 column heater/cooler HT column heater B-12 column manager sample manager...
Page 509 3-38 lamp, TUV detector 6-221 reference energies, recording 3-38 primary head plunger, binary refresh system control solvent manager 6-97 refreshing, ACQUITY UPLC system puncture needle holder 6-142 3-12 sample loop 6-145 regulator, backpressure 2-23 sample syringe 6-149 removing...
Page 510 column manager 4-12 needle assembly TUV detector 4-10 installing 6-135 restrictors 3-32 removing 6-131 routine maintenance seal, characterizing 3-21 3-22 run LED 4-11 7-14 7-17 7-21 6-140 running volume, characterizing 3-22 HPLC methods 3-42 6-141 system test mix, Empower 5-10 washing 3-20 Z-axis, calibrating...
Page 511 wash syringes, replacing 6-152 solvent manager head washing needle 3-19 removing 6-53 6-70 XYZ mechanism, calibrating 2-67 solvent manager, see binary solvent sample organizer manager air filters, cleaning 6-225 solvents displaying plate information 3-35 buffered fuses, replacing 6-227 changing information window 3-36 clean initiating communications to...
Page 512 MassLynx software 4-14 system pressure Empower software system verification MassLynx software 4-13 TUV detector 3-37 teach block 2-67 starting up, ACQUITY UPLC system test methods, creating 3-13 tests, startup startup control tetrahydrofuran startup diagnostic tests 3-37 C-10 C-11 C-13...
Page 513 B-17 wash solvents, selecting 3-15 flow cell B-19 wash syringes, replacing 6-152 operational B-18 washing, sample manager needle 3-20 starting 3-37 waste line positioning 2-32 troubleshooting 7-24 7-27 – water, as solvent verifying 3-38 Waters Technical Service, contacting Index-13...
Page 514 wavelength cutoffs C-11 display mobile phase absorbances C-12 selection C-11 weak needle wash connection 2-27 XYZ mechanism, calibrating 2-67 Index-14...