Waters TQ Detector Operator’s Guide - Xi'an University ...jg.xauat.edu.cn/labs/hjgch/files/pdf/2014/20141216105217_9573.pdf · 1 Waters TQ Detector Overview This chapter describes

Waters TQ DetectorQuick Start Guide

71500126803/Revision A

Copyright © Waters Corporation 2007.All rights reserved.

Copyright notice

© 2007 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA AND IRELAND. ALL RIGHTS RESERVED. THIS DOCUMENT OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.

The information in this document is subject to change without notice and should not be construed as a commitment by Waters Corporation. Waters Corporation assumes no responsibility for any errors that may appear in this document. This document is believed to be complete and accurate at the time of publication. In no event shall Waters Corporation be liable for incidental or consequential damages in connection with, or arising from, its use.

Waters Corporation34 Maple StreetMilford, MA 01757USA

TrademarksConnections Insight, ESCi, and Waters are registered trademarks of Waters Corporation. ACQUITY UPLC, IntelliStart, IonSABRE, MassLynx, T-Wave, UPLC, and ZSpray are trademarks of Waters Corporation.

Other trademarks or registered trademarks are the sole property of their respective owners.

Customer commentsWaters’ Technical Communications department invites you to tell us of any errors you encounter in this document or to suggest ideas for otherwise improving it. Please help us better understand what you expect from our documentation so that we can continuously improve its accuracy and usability.

We seriously consider every customer comment we receive. You can reach us at [email protected].

Waters Corporation34 Maple StreetMilford, MA 01757USA

iii

Safety considerations

Some reagents and samples used with Waters® instruments can pose chemical, biological, and radiological hazards. Be sure you are aware of the potentially hazardous effects of all substances you work with. Always observe Good Laboratory Practice (GLP) guidelines, published by the U.S. Food and Drug Administration, and consult your organization’s safety representative for guidance.

When you develop methods, follow the “Protocol for the Adoption of Analytical Methods in the Clinical Chemistry Laboratory,” American Journal of Medical Technology, 44, 1, pages 30–37 (1978). This protocol addresses good operating procedures and the techniques necessary to validate system and method performance.

Instrument-specific safety considerations

Solvent leakage hazard

The source exhaust system is designed to be robust and leak-tight. Waters recommends you perform a hazard analysis, assuming a maximum leak into the laboratory atmosphere of 10% HPLC eluate.

Warning: • To confirm the integrity of the source exhaust system, the

source seals must be renewed at intervals not exceeding one year.

• The source seals can withstand exposure only to specific solvents, see Appendix C in the Waters TQ Detector Operator’s Guide. If you intend to use solvents other than those listed, you must first determine whether those solvents are compatible with the composition of the seals.

iv

Flammable solvents hazard

Never let the nitrogen supply pressure fall below 400 kPa (4 bar, 58 psi) during analyses that require flammable solvents. Connect to the LC output with a gas-fail connector to stop the LC solvent if the nitrogen supply fails.

High temperature hazard

Waters TQ Detector high temperature hazard:

Warning: Where significant quantities of flammable solvents are involved, the source must receive a continuous flow of nitrogen to prevent possible ignition within that enclosed space.

Warning: The source enclosure can be hot. To avoid burn injuries, avoid touching the source enclosure when operating or servicing the TQ Detector.

Source enclosure assembly

v

High voltage hazard

Safety advisoriesConsult Appendix A for a comprehensive list of warning and caution advisories.

Warning: • To avoid electric shock, do not remove the TQ Detector’s protective

panels. The components they cover are not user-serviceable.• To avoid non-lethal electric shock, any equipment connected to the

ESI and IonSABRE™ APCI probes must be grounded.• When the TQ Detector is in Operate mode, certain external surfaces

can conduct high voltages. To avoid non-lethal electric shock, make sure the instrument is in Standby mode before touching areas marked with this high voltage warning symbol.

vi

Operating this device

When operating this device, follow standard quality control procedures and the guidelines presented in this section.

Symbols

Intended useThe Waters Tandem Quadrupole (TQ) Detector is intended to be used as a research tool to deliver authenticated mass measurement in both MS and MS/MS modes.

The Waters TQ Detector can be used for general In Vitro Diagnostic applications, only by professionally trained and qualified laboratory personnel.

Symbol Definition

Authorized Representative of the European Community

The CE symbol serves as confirmation of the conformity of a product with all European Community directives applicable to that product.

For in vitro diagnostic use.

The Waters TQ Detector is CE-marked according to the European Union In Vitro Diagnostic Device Directive 98/79/EC.

EC REP

IVD

vii

CalibrationTo calibrate LC systems, follow acceptable calibration methods using at least five standards to generate a standard curve. The concentration range for standards should cover the entire range of quality-control samples, typical specimens, and atypical specimens.

To calibrate mass spectrometers, consult the calibration section of the operator’s guide of the instrument you are calibrating.

Quality controlRoutinely run three quality-control samples that represent subnormal, normal, and above-normal levels of a compound. Ensure that quality-control sample results fall within an acceptable range, and evaluate precision from day to day and run to run. Data collected when quality control samples are out of range might not be valid. Do not report these data until you are certain that the instrument performs satisfactorily.

When analyzing samples from a complex matrix such as soil, tissue, serum/plasma, whole blood, etc., note that the matrix components can adversely affect LC/MS results, enhancing or suppressing ionization. To minimize these matrix effects, Waters recommends you adopt the following measures:

• Prior to the instrumental analysis, use appropriate sample pretreatment such as protein precipitation, liquid/liquid extraction (LLE), or solid phase extraction (SPE) to remove matrix interferences.

• Whenever possible, verify method accuracy and precision using matrix-matched calibrators and QC samples.

• Use one or more internal standard compounds, preferably isotopically-labeled analytes.

viii

IVD authorized representative information

IVD authorized representative

Waters Corporation (Micromass UK Limited) is registered in the United Kingdom with the Medicines and Healthcare Products Regulatory Agency (MHRA) at market Towers, 1 Nine Elms Lane, London, SW8 5NQ. The reference number is IVD000167.

Waters Corporation (Micromass UK Ltd.)Floats RoadWythenshaweManchester M23 9LZUnited Kingdom

Telephone: +44-161-946-2400

Fax: +44-161-946-2480

Contact: Quality manager

ix

x

Table of Contents

Safety considerations .......................................................................................... iv Instrument-specific safety considerations......................................................... iv Flammable solvents hazard ................................................................................ v High temperature hazard.................................................................................... v High voltage hazard............................................................................................ vi Safety advisories ................................................................................................. vi

Operating this device ......................................................................................... vii Symbols ............................................................................................................. vii Intended use...................................................................................................... vii Calibration ....................................................................................................... viii Quality control ................................................................................................. viii

IVD authorized representative information ................................................. ix IVD authorized representative .......................................................................... ix

1 Waters TQ Detector Overview ............................................................ 1-1

Overview ............................................................................................................. 1-2 Waters TQ Detector ......................................................................................... 1-2 ACQUITY TQD UPLC/MS system ................................................................. 1-4 MassLynx mass spectrometry software and data system ............................. 1-5 ACQUITY UPLC Console................................................................................ 1-6

Ionization techniques and source probes ................................................... 1-7 Electrospray ionization (ESI) .......................................................................... 1-7 Combined electrospray ionization and atmospheric pressure chemical ionization

(ESCi) ......................................................................................................... 1-7 Atmospheric pressure chemical ionization..................................................... 1-8 Atmospheric pressure photoionization ........................................................... 1-8

Table of Contents xi

Ion optics ............................................................................................................. 1-9

MS operating modes ....................................................................................... 1-10

MS/MS operating modes ................................................................................ 1-11 Product (daughter) ion mode......................................................................... 1-11 Precursor (parent) ion mode......................................................................... 1-12 Multiple reaction monitoring mode .............................................................. 1-12 Constant neutral loss mode........................................................................... 1-13

Sample inlet ...................................................................................................... 1-14

Vacuum system ................................................................................................ 1-14

Rear panel ......................................................................................................... 1-15

IntelliStart fluidics system overview ......................................................... 1-16

IntelliStart fluidics system operation ........................................................ 1-17 Operating the IntelliStart fluidics system from the ACQUITY UPLC

Console ..................................................................................................... 1-17 Operating the IntelliStart fluidics system from the Tune window............. 1-18 Programming the MS method to operate the IntelliStart fluidics system . 1-18

2 Preparing the Waters TQ Detector for Operation ......................... 2-1

Starting the TQ Detector ................................................................................. 2-2 Configuring IntelliStart................................................................................... 2-6 Verifying the instrument’s state of readiness ................................................ 2-6 Tuning and calibration information ............................................................... 2-6 Running the TQ Detector at high flow rates .................................................. 2-7 Monitoring the TQ Detector LEDs.................................................................. 2-8

Preparing the IntelliStart fluidics system .................................................. 2-9 Installing the solvent manifold drip tray ....................................................... 2-9 Installing the reservoir bottles...................................................................... 2-10 Diverter valve positions................................................................................. 2-11 Purging the infusion syringe......................................................................... 2-13

Rebooting the TQ Detector ........................................................................... 2-14 Rebooting the TQ Detector by pressing the reset button ............................ 2-14

xii Table of Contents

Shutting down the TQ Detector .................................................................. 2-15 Putting the TQ Detector in Standby mode for overnight shutdown ........... 2-15 Complete TQ Detector shutdown.................................................................. 2-16 Emergency TQ Detector shutdown ............................................................... 2-17

3 ESI and ESCi Modes of Operation ..................................................... 3-1

Introduction ....................................................................................................... 3-2

Installing the ESI probe .................................................................................. 3-2 Required materials .......................................................................................... 3-2

Installing the corona pin ................................................................................. 3-5 Required materials .......................................................................................... 3-5

Optimizing the ESI probe for ESCi operation ........................................... 3-7 Required materials .......................................................................................... 3-7

Removing the corona pin ................................................................................ 3-9 Required materials .......................................................................................... 3-9

Removing the ESI probe ............................................................................... 3-11 Required materials ........................................................................................ 3-11

4 Operating the Waters TQ Detector .................................................... 4-1

Setting-up the instrument .............................................................................. 4-2 Required materials .......................................................................................... 4-2

Performing a sample tune .............................................................................. 4-7 Required materials .......................................................................................... 4-7

Developing experiment methods .................................................................. 4-9 Required materials .......................................................................................... 4-9

Verifying the system using System QC ...................................................... 4-11

A Safety Advisories .................................................................................. A-1

Warning symbols ............................................................................................... A-2 Task-specific hazard warnings........................................................................ A-2 Warnings that apply to particular instruments, instrument components, and

sample types............................................................................................... A-3

Table of Contents xiii

Caution symbol .................................................................................................. A-5

Warnings that apply to all Waters instruments ......................................... A-5

Electrical and handling symbols ................................................................. A-13 Electrical symbols .......................................................................................... A-13 Handling symbols .......................................................................................... A-14

xiv Table of Contents

1 Waters TQ Detector Overview

This chapter describes the instrument, including its controls and gas and plumbing connections.

Contents:

Topic Page

Overview 1-2

Ionization techniques and source probes 1-7

Ion optics 1-9

MS operating modes 1-10

MS/MS operating modes 1-11

Sample inlet 1-14

Vacuum system 1-14

Rear panel 1-15

IntelliStart fluidics system overview 1-16

IntelliStart fluidics system operation 1-17

1-1

Overview

Waters TQ Detector

The Waters® TQ Detector is a tandem quadrupole, atmospheric pressure ionization (API) mass spectrometer. Designed for routine UPLC™/MS/MS analyses in quantitative and qualitative applications, it can operate at fast acquisition speeds compatible with ultra performance LC.

Waters provides these ion sources with the instrument as standard equipment:

• ZSpray™ (dual orthogonal sampling) interface.

• Multi-mode ESCi® ionization switching for atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI).

Optional ionization modes are IonSABRE™ APCI and APPI (atmospheric pressure photoionization).

For TQ Detector specifications, see the Waters TQ Detector Site Preparation Guide.

Waters TQ Detector:

TP02592

1-2 Waters TQ Detector Overview

Waters TQ Detector with doors open:

IntelliStart technology

IntelliStart™ technology monitors LC/MS/MS performance and reports when the TQ Detector is ready for use.

The IntelliStart software automatically tunes and mass calibrates the TQ Detector and displays performance readbacks. Integrated with MassLynx™ mass spectrometry software and ACQUITY UPLC™ Console software, IntelliStart enables simplified set-up of the system for use in routine analytical and open access applications.

The IntelliStart fluidics system is built into the TQ Detector. It delivers sample directly to the MS probe from the LC column or from two integral reservoirs. The integral reservoirs can also deliver sample through direct or combined infusion so that you can optimize instrument performance at analytical flow rates.

See also: The TQ Detector online Help for further details of IntelliStart.

NEBULIZER

DESOLVATION

TP02627

POWER OPERATE

HVHV

PROBEPROBE

Overview 1-3

ACQUITY TQD UPLC/MS systemNote: The Waters TQ Detector is designed for compatibility with the ACQUITY UPLC system; if you are not using an ACQUITY UPLC system, refer to the documentation relevant to the LC system being used.

The ACQUITY TQD UPLC/MS system includes an ACQUITY UPLC system and the Waters TQ Detector.

ACQUITY UPLC system

The ACQUITY UPLC system includes a binary solvent manager, sample manager, column heater, optional sample organizer, optional detectors, and a specialized ACQUITY UPLC column. Waters MassLynx mass spectrometry software controls the system.

See also:

• ACQUITY UPLC System Operator’s Guide

• Controlling Contamination in LC/MS Systems (part number 715001307). You can find this document on http://www.waters.com; click Services and Support and then Support Center.


Waters ACQUITY TQD:

MassLynx mass spectrometry software and data systemMassLynx is a high-performance mass spectrometry application that acquires, analyzes, manages, and distributes ultra-violet (UV), evaporative light scattering, analog, and mass spectrometry data.

MassLynx software permits these major operations:

• Configuring the instrument.

• Creating LC inlet and MS/MS methods that define operating parameters for a run.

• Using IntelliStart software to tune and mass calibrate the TQ Detector.

• Running samples.

• Monitoring the run.

• Acquiring data.

• Processing data.

TP02597

Sample organizer (optional)

Solvent tray

Column heater

TQ Detector

Sample managerBinary solvent manager

Overview 1-5

• Reviewing data.

• Printing data.

See also: MassLynx 4.1 user documentation and online Help for more information on installing and using MassLynx software.

ACQUITY UPLC ConsoleThe ACQUITY UPLC Console is a software application with which you configure settings, monitor performance, run diagnostic tests, and maintain the system and its modules. The ACQUITY UPLC Console functions independently of MassLynx and does not recognize or control the data system.

See also: ACQUITY UPLC System console online Help for details of the TQ Detector.

TQ Detector ACQUITY UPLC Console page:


Ionization techniques and source probes

Electrospray ionization (ESI)In electrospray ionization (ESI), a strong electrical charge is given the eluent as it emerges from a nebulizer. The droplets that compose the resultant aerosol undergo a reduction in size (solvent evaporation). As solvent continues to evaporate, the charge density increases until the droplet surfaces eject ions (ion evaporation). The ions can be singly or multiply charged. The multiply charged ions are of particular interest because the TQ Detector separates them according to their mass-to-charge ratios (m/z), permitting the detection of high-molecular-weight compounds.

The instrument can accommodate eluent flow rates of up to 1 mL/min.

Combined electrospray ionization and atmospheric pressure chemical ionization (ESCi)

Combined electrospray ionization and atmospheric pressure chemical ionization (ESCi) is supplied as standard equipment on the TQ Detector. In ESCi, the standard ESI probe is used in conjunction with a corona pin to allow alternating acquisition of ESI and APCI ionization data, facilitating high throughput and wider compound coverage.

Ionization techniques and source probes 1-7

ESCi mode:

See also: “Electrospray ionization (ESI)” on page 1-7.

Atmospheric pressure chemical ionizationA dedicated high performance atmospheric pressure chemical ionization (APCI) probe is offered as an option.

See also: The Waters TQ Detector Operator’s Guide for full details.

Atmospheric pressure photoionizationAtmospheric pressure photoionization (APPI) is offered as an option. It uses photons generated by a krypton-discharge ultra-violet (UV) lamp (∼ 10.2 eV) to produce sample ions from vaporized LC eluent.

TP02695

Corona pin

Sample cone tip


Ion optics

The TQ Detector’s ion optics operate as follows:

• Samples from the LC or Intellistart fluidics system are introduced at atmospheric pressure into the ionization source.

• The ions pass through the sample cone into the vacuum system.

• The ions pass through the transfer optics to the first quadrupole where they are filtered according to their mass-to-charge ratio (m/z).

• The mass-separated ions pass into the T-Wave™ collision cell where they either undergo collision-induced dissociation (CID) or pass to the second quadrupole. Any fragment ions are then mass-analyzed by the second quadrupole.

• The transmitted ions are detected by the photomultiplier detection system.

• The signal is amplified, digitized, and sent to the MassLynx mass spectrometry software.

Ion optics overview:

Sample cone

Isolation valve

T-Wave collision cell

Z-Spray ion source Quadrupole 1 (MS1)

Quadrupole 2 (MS2)

Detector

Conversion dynodeSample inlet Transfer optics

Ion optics 1-9

MS operating modes

The following table shows the MS operating modes.

The MS1 mode, in which MS1 is used as the mass filter, is the most common and most sensitive method of performing MS analysis. It is directly analogous to using a single-quadrupole mass spectrometer.

The MS2 mode of operation is used, with collision gas present, when switching rapidly between MS and MS/MS operation (for example, in survey scan mode). It also provides a useful tool for instrument tuning and calibration before MS/MS analysis and for fault diagnosis.

The selected ion recording (SIR) mode of operation is used as a quantitation mode when no suitable fragment ion can be found to perform a more specific multiple reaction monitoring (MRM) analysis.

MS operating modes :

Operating mode MS1 Collision cell MS2

MS1 Resolving (scanning)

Pass all masses

MS2 Pass all masses Resolving (scanning)

SIR Resolving (static) Pass all masses


MS/MS operating modes

The following table shows the MS/MS operating modes.

Product (daughter) ion modeProduct ion mode is the most commonly used MS/MS operating mode. An ion of interest is selected for fragmentation in the collision cell, thus yielding structural information.

Product ion mode:

Typical applications

• Structural elucidation (for example, peptide sequencing)

MS/MS operating modes :

Operating mode MS1 Collision cell MS2

Product (daughter) ion spectrum

Static (at precursor mass)

Pass all masses Scanning

Precursor (parent) ion spectrum

Scanning Static (at product mass)

MRM Static (at precursor mass)

Static (at product mass)

Constant neutral loss spectrum

Scanning (synchronized with MS2)

Scanning (synchronized with MS1)

MS1Static (at precursor mass)

MS2Scanning

Collision cellPass all masses


• Method development for MRM screening studies:

– Identifying product ions for use in MRM transitions.

– Optimizing CID tuning conditions to maximize the yield of a specific product ion to be used in MRM analysis.

Precursor (parent) ion mode

Precursor ion mode:

Typical application

You typically use the precursor ion mode for structural elucidation–that is, to complement or confirm product scan data–by scanning for all the precursors of a common product ion.

Multiple reaction monitoring modeMRM mode is a highly selective MS/MS equivalent of SIR. As both MS1 and MS2 are static, greater dwell time on the ions of interest is allowed, and therefore better sensitivity compared to scanning MS/MS. This is the most commonly used acquisition mode for quantitative analysis, allowing the compound of interest to be isolated from the chemical background noise.

MS1Scanning

MS2Static (at product mass)



Multiple reaction monitoring mode:

Typical application

You typically use MRM to quantify known analytes in complex samples:

• Drug metabolite and pharmacokinetic studies.

• Environmental, for example, pesticide and herbicide analysis.

• Forensic or toxicology, for example, screening for target drugs in sport.

• MRM does not produce a spectrum because only one transition is monitored at a time. As in SIR, a chromatogram is produced.

Constant neutral loss modeConstant neutral loss mode detects the loss of a specific neutral fragment or functional group from an unspecified precursor or precursors.

The scans of MS1 and MS2 are synchronized. When MS1 transmits a specific precursor ion, MS2 “looks” to see whether that precursor loses a fragment of a certain mass. If it does, the loss registers at the detector.

In constant neutral loss mode, the spectrum shows the masses of all precursors that actually lost a fragment of a certain mass.

MS1Static (at precursor mass)

MS2Static (at product mass)



Constant neutral loss mode:

Typical application

You typically use constant neutral loss mode to screen mixtures for a specific class of compound that is characterized by a common fragmentation pathway, indicating the presence of compounds containing a common functional group.

Sample inlet

Either of two methods delivers solvent and sample to the installed probe:

• An LC system, which delivers the eluent from an LC analysis.

• IntelliStart fluidics system, which uses on-board solutions to automate instrument optimization. You can deliver solutions by direct or combined infusion.

Vacuum system

An external roughing (rotary vane) pump and an internal split- flow turbomolecular pump combine to create the source vacuum. The turbomolecular pump evacuates the analyzer and ion transfer region.

Vacuum leaks and electrical or vacuum pump failures cause vacuum loss, which protective interlocks guard against. The system monitors turbomolecular pump speed and continuously measures vacuum pressure with a built-in Pirani gauge. The gauge also serves as a switch, discontinuing operation when it senses vacuum loss.

A vacuum isolation valve isolates the source from the mass analyzer, allowing routine source maintenance without venting.

MS1Scanning

(synchronized with MS2)


MS2Scanning

(synchronized with MS1)


Rear panel

The following figure shows the rear panel locations of the connectors used to operate the TQ Detector with external devices.

TQ Detector rear panel:

�

VACUUM

Waters Corporation34 Maple Street Milford, MA 01757 U.S.A.

�Serial Number

ACN 065444751

VACUUM

SOURCE VENT

API GAS

6.9 BAR MAXIMUM

V~ 200-240

Hz 50-60

VA 1200

RS 232

ETHERNET

PUMP

COLLISION

GAS

1.0 BAR

MAXIMUM

IVD

+

-Not Used

Switch 4

Ground

Ground

Stop Flow

Switch 4

Ground

Switch 3

Event

Ground

Inject StartOut

Out Out

In

1

2

3

4

5

6

78

9

10

1

2

3

4

5

6

78

9

10

Out

InAnalog+

-

+

-

+

-

+

-

+

-

Event inputs and outputs

Shielded Ethernet

Roughing pump relay switch

Source vacuum

Power cord

Source vent

Turbo vacuum

Nitrogen inlet

Collision cell gas inlet

Rear panel 1-15

IntelliStart fluidics system overview

The IntelliStart fluidics system is built into the TQ Detector. The system delivers sample directly to the MS probe in one of two ways:

• From the LC column

• From two integral reservoirs.

Tip: The integral reservoirs can also deliver sample through direct or combined infusion to enable optimization at analytical flow rates.

The IntelliStart system incorporates a multi-position valve with these attributes:

• An input connection from an external LC column.

• An input connection from the TQ Detector’s infusion syringe. (The TQ Detector’s infusion syringe is also connected to two reservoirs, A and B. In the software, you specify which reservoir to draw from.)

• An output connection to the TQ Detector’s probe.

• An output connection to a waste line.

LC flow:

Column

LC

LC Waste

Probe

Syringe

Idle

Reservoir A Reservoir B

Off

Off

B

A


IntelliStart fluidics system operation

Control of solvent and sample delivery during auto-tuning, auto-calibration, and method development is automatically performed by the software.

IntelliStart configuration requirements can be set in the system console. You can edit the parameters, frequency, and extent of the automation you want IntelliStart to perform.


Operating the IntelliStart fluidics system from the ACQUITY UPLC Console

To operate the IntelliStart fluidics system from the Instrument Console:

1. In the MassLynx window, click MS Console.

2. In the ACQUITY UPLC Console system tree, expand TQ Detector.

3. Click Interactive Display.

4. In the ACQUITY UPLC Console window, click the current flow rate.

5. In the Select Flow Rate dialog box, enter a new flow rate.


6. Click OK.

7. Click Reservoir A or B.

8. In the Select Reservoir dialog box, click A or B.

9. Click OK.

Tip: If the selected reservoir is different from the current reservoir, the system purges as the reservoir changes.

10. Click the current diverter valve position label.

11. In the Select a Flow State dialog box, select the required flow state.

12. Click OK.

13. Click to start.

Tip: A status indicates the amount of fluid in the syringe and the amount of time remaining before the fluid empties. When the syringe is empty the system becomes idle.

14. Click to refill the syringe or to purge the system.

15. Click to stop the current action.

Operating the IntelliStart fluidics system from the Tune window

To operate the IntelliStart fluidics system from the Tune window:

1. In the MassLynx window, click MS Tune.

2. In the TQ Detector Tune window, click the Fluidics page.

3. Set the Flow Control parameters according to the instructions in the MassLynx online Help.

Programming the MS method to operate the IntelliStart fluidics system

In the MS method, you can program the operation of the system’s multi-position valve to infuse sample during a run. The valve can also divert LC flow to waste as a timed event.


To program the MS method:

1. In the MassLynx window, click MS Method.

2. In the MS Methods window, click Options > Method Events.

3. In the Method events dialog box, select the appropriate flow state, as shown in the following table.

Tip: At instrument power-up, the LC state is waste.

See also: MassLynx online Help topic “Advanced Methods and Events”.

Flow states :

State LC Syringe

LC TQ probe Waste

Combined TQ probe TQ probe

Infusion Waste TQ Probe

Waste Waste Waste

No change No change No change



2 Preparing the Waters TQ Detector for Operation

This chapter describes how to start and shut-down the TQ Detector.

Contents:

Topic Page

Starting the TQ Detector 2-2

Preparing the IntelliStart fluidics system 2-9

Rebooting the TQ Detector 2-14

Shutting down the TQ Detector 2-15

2-1

Starting the TQ Detector

Note: The Waters TQ Detector is designed for compatibility with the ACQUITY UPLC system; if you are not using an ACQUITY UPLC system, refer to the documentation relevant to the system being used.

Starting the TQ Detector entails powering-on the ACQUITY workstation, logging into the workstation, powering-on the TQ Detector and all the other ACQUITY instruments, and starting the MassLynx software.

Requirement: You must power-on and log in to the ACQUITY workstation first to ensure that it obtains the IP addresses of the system instruments.

To start the TQ Detector:

1. Ensure the nitrogen supply is connected to the instrument’s API gas connection.

Note: The nitrogen must be dry and oil-free, with a purity of at least 95%. Regulate the supply at 600 to 690 kPa (6.0 to 6.9 bar, 90 to 100 psi).

See also: Figure titled “TQ Detector rear panel” on page 1-15.

2. Ensure that the collision gas supply is connected to the TQ Detector.

Requirement: The collision gas is argon; it must be dry and of high purity (99.9%). Regulate the supply at 50 kPa (0.5 bar, 7 psi).

3. Power-on the ACQUITY UPLC system workstation, and log in before powering-on the other instruments.

Caution: Using incompatible solvents can cause severe damage to the instrument.• Refer to Appendix C of the Waters TQ Detector Operator’s Guide for

TQ Detector solvent information.• Refer to Appendix C of the ACQUITY UPLC System Operator’s

Guide for more information on solvent compatibility with ACQUITY.

Warning: During analyses that require flammable solvents, to avoid ignition of the solvents, never let the nitrogen supply pressure fall below 400 kPa (4 bar, 58 psi).

2-2 Preparing the Waters TQ Detector for Operation

4. Press the power switch on the top, left-hand side of the TQ Detector and ACQUITY instruments. Each system instrument “beeps” and runs a series of startup tests.

5. Allow 3 minutes for the embedded PC to initialize. An audible alert sounds when the PC is ready.

The power and status LEDs change as follows:

• Each system instrument’s power LED shows green.

• During initialization, the binary solvent manager’s and sample manager’s status LED flashes green.

• After the instruments are successfully powered-on, all power LEDs show steady green. The binary solvent manager’s flow LED, the sample manager’s run LED, and the TQ Detector’s operate LED remain off.

6. Start MassLynx. You can monitor the ACQUITY console for messages and LED indications.

7. In the MassLynx main window’s lower-left corner, click IntelliStart.

Result: TQ Detector console IntelliStart appears. The TQ Detector is in Standby mode.


8. Click Control > Pump to start the roughing pump. The operate LED remains off.

Tip: There is a 20-second delay, during which the turbopump is starting, before the roughing pump starts. IntelliStart displays “Instrument in standby”.


9. Click Resolve or Operate to put the TQ Detector into Operate mode. When the TQ Detector is in good operating condition, IntelliStart displays “Ready”.

Tip: Clicking Resolve should prepare the system for operation, putting the TQ Detector into Operate mode. If clicking Resolve fails to put the instrument into Operate mode, IntelliStart displays corrective actions.


Configuring IntelliStart

To configure IntelliStart:

1. In the ACQUITY UPLC Console system tree, click TQ Detector.

2. Click Configure > IntelliStart Configuration.

3. In the IntelliStart Configuration dialog box, in the Checks list, select the checkboxes for the items you want checked during TQ Detector startup. Clear the checkboxes of items you do not want checked.

Tip: To display detailed information for an item, highlight it and click Properties.

4. Click Apply > OK.

Verifying the instrument’s state of readinessWhen the TQ Detector is in good operating condition, the power LED shows constant green and the operate LED is off. You can view any error messages in IntelliStart.

To access IntelliStart:

1. In the ACQUITY UPLC Console system tree, select TQ Detector.

2. In the TQ Detector information window, click IntelliStart.

Tuning and calibration informationThe TQ Detector must be tuned and calibrated prior to use, tasks normally performed from IntelliStart.

See also: The online Help topic “Instrument Setup” and Chapter 4, “Operating the Waters TQ Detector”.


Running the TQ Detector at high flow ratesACQUITY UPLC is run at high flow rates. To optimize desolvation, and thus sensitivity, the ACQUITY TQD system should be run at appropriate gas flows and desolvation temperatures. IntelliStart automatically sets these when you enter a flow rate, according to the following table.

Note: Under low ambient temperature, high moisture, and high flow rate conditions, condensation can occur in the instrument’s source.

Flow rate versus temperature and gas flow :

Flow rate (mL/min) Source temp (°C) Desolvation

temp (°C)Desolvation gas flow (L/hr)

0.000 to 0.100 120 250 500

0.101 to 0.300 120 350 600

0.301 to 0.500 150 400 800

>0.500 150 450 900


Monitoring the TQ Detector LEDsLight-emitting diodes on the TQ Detector indicate its operational status.

Power LED

The power LED, to the top, left-hand side of the TQ Detector’s front panel, indicates when the TQ Detector is powered-on or powered-off.

Operate LED

The operate LED, to the right of the power LED, indicates the operating condition.

Operate LED indications :

LED mode and color Description

Off The instrument is in Standby mode with high voltages and heaters inoperative.

Constant green The instrument is in Operate mode with heaters and high voltages operating.

Flashing green Indicates the system is waiting for an instrument component to reach operational conditions. The LED could, for example, flash while the system reaches a programmed temperature set point or vacuum level. The instrument is in working order and Operate mode but not yet ready to acquire.

Flashing red Indicates that the system stopped due to error and is no longer processing any samples or other maintenance requests. Information regarding the error appears in the data system's control panel or the ACQUITY UPLC Console software.

Constant red for 3 minutes

The instrument is waiting for the instrument to initialize.

Constant red for more than 3 minutes

Indicates a serious system failure that prevents further operation. Power-off the instrument, and then power-on. If the LED is still steady red for more than 3 minutes, contact your Waters service representative.


Preparing the IntelliStart fluidics system

See also: Appendix B of the Waters TQ Detector Operator’s Guide.

Installing the solvent manifold drip tray

Required materials

Chemical-resistant, powder-free gloves

To install the solvent manifold drip tray:

Install the solvent manifold drip tray as shown below:

Warning: The solvent manifold drip tray can be contaminated with biohazardous and/or toxic materials. Always wear chemical-resistant, powder-free gloves while performing this procedure.

TP02685


Installing the reservoir bottles

Required materials

Chemical-resistant, powder-free gloves

To install the reservoir bottles:

1. Remove the reservoir bottle caps.

2. Screw the reservoir bottles onto the TQ Detector as shown below.

Warning: The reservoir bottles can be contaminated with biohazardous and/or toxic materials. Always wear chemical-resistant, powder-free gloves while performing this procedure.

TP02630


Diverter valve positions

Column and syringe in home position after power-up

After power-up, the flow path between the column and waste is open. The syringe is empty, and the flow path between it and waste is open.

LC position

In the LC position, the flow path between the LC and probe is open, and the flow path between the syringe and waste is also open.

Column

Waste

LC Waste

Probe

Syringe

Idle


Off

Off

B

A

Column

LC

LC Waste

Probe

Syringe

Idle


Off

Off

B

A


Infusion position in infusion mode

Combined position with LC flow and syringe in idle mode

Column

Infusion

LC Waste

Probe

Syringe


Off

On

B

A

Infusion

Column

Combined

LC Waste

Probe

Syringe


Off

Off

B

A

Idle


Waste position

In the waste position, both the LC flow and the infusion syringe flow are directed to waste. The syringe mode can be only static or dispensing (that is, never drawing).

Purging the infusion syringeWhenever you replace a solution bottle, you should purge the infusion syringe with the solution that you are going to use next.

Tip: Depending on the solutions used, the IntelliStart fluidics system can require more then one purge cycle to minimize carryover.

To purge the infusion syringe:



3. Select the required solution reservoir.

4. Click to purge the system.

Tip: System purge takes approximately 2 minutes.

Column

Waste

LC Waste

Probe

Syringe


Off

Off

B

A

Idle


Rebooting the TQ Detector

Sometimes you might need to reboot the TQ Detector:

• When the Tune window fails to respond.

• When MassLynx fails to initialize.

• Immediately following software download.

Rebooting the TQ Detector by pressing the reset buttonTip: The reset button shuts down the electronics momentarily and causes the TQ Detector to reboot.

To reboot the TQ Detector by pressing the reset button:

1. Open the TQ Detector’s front left door.

2. Press the red reset button on the top, left-hand side of the instrument.

TP02687

Reset button


Shutting down the TQ Detector

Recommendation: You should always leave the TQ Detector in Operate mode except when performing routine maintenance. It is not necessary to switch to Standby mode. However, shutting down the TQ Detector is acceptable provided that you consider warm-up time on restarting. If you do shut down the TQ Detector, refer to the instructions in this section.

Tip: Set system shutdown parameters in the Shutdown Editor. Consult the MassLynx online Help for more information.

Putting the TQ Detector in Standby mode for overnight shutdownYou might want to shut down the instrument for a relatively brief period, like overnight, while maintaining the LC flow.

To put the TQ Detector in Standby mode:

1. In the ACQUITY UPLC Console, click to stop the LC flow or, if column flow is required, divert the LC flow to waste as follows:

a. In the ACQUITY UPLC Console system tree, expand TQ Detector.

b. Click Interactive Display.

c. Click the current diverter valve position label.

d. In the Select a Flow State dialog box, select Waste.

e. Click OK.

2. Click Standby to shut off heaters and voltages.

Tip: You can create a method to stop the gas flow or lower the temperature. See the MassLynx online Help for more information on creating methods.

Caution: Buffers left in the system can precipitate and damage instrument components.


Complete TQ Detector shutdown

To completely shut down the TQ Detector:

1. Put the TQ Detector in Standby mode.

See also: “Putting the TQ Detector in Standby mode for overnight shutdown” on page 2-15.

2. In the ACQUITY UPLC Console, click API Gas .

3. Click Col Gas .

4. Click Control > Vent.

Result: The turbomolecular pump is switched off. When the turbomolecular pump runs down to half its normal operating speed, the vent valve opens and the instrument is automatically vented. The operate LED changes from green to red and then turns off.

5. Exit MassLynx.

6. After the roughing pump shuts off, operate the power button (on the front of the instrument) to power-off the TQ Detector.

7. Disconnect the power cable from the back of the TQ Detector.

8. Power-off all other instruments and the workstation.

Note: The fans inside some instruments run continuously, even after you power-off the instruments.

Warning: The TQ Detector’s power switch does not isolate the instrument from the main power supply. To isolate the instrument, you must disconnect the power cable from the back of the instrument.


Emergency TQ Detector shutdown

To shut down the TQ Detector in an emergency:

1. Operate the power button on the front of the TQ Detector.

2. Disconnect the power cable from the back of the TQ Detector.

Warning: The TQ Detector’s power switch does not isolate the instrument from the main power supply. To isolate the instrument, you must disconnect the power cable from the back of the instrument.

Caution: Data can be lost during an emergency shutdown.



3 ESI and ESCi Modes of Operation

This chapter describes how to prepare the TQ Detector for the following modes of operation:

• ESI (electrospray ionization)

• ESCi (combined electrospay and atmospheric pressure chemical ionization)

If your system uses APCI mode, refer to Chapter 6 of the Waters TQ Detector Operator’s Guide.

Contents:

Topic Page

Introduction 3-2

Installing the ESI probe 3-2

Installing the corona pin 3-5

Optimizing the ESI probe for ESCi operation 3-7

Removing the corona pin 3-9

Removing the ESI probe 3-11

3-1

Introduction

The ESI and ESCi ionization mode options use the standard ESI probe that is fitted to the instrument when it is shipped from the factory. For ESCi operation, the corona pin is used in conjunction with the ESI probe. The following sections explain how to install and remove the ESI probe and corona pin.

See also:

• “Electrospray ionization (ESI)” on page 1-7.

• Table titled “Combined electrospray ionization and atmospheric pressure chemical ionization (ESCi)” on page 1-7.

Installing the ESI probe

Required materialsChemical-resistant, powder-free gloves

To install the ESI probe:

1. Prepare the instrument for working on the source.

See also: Chapter 5 of the Waters TQ Detector Operator’s Guide.

2. Open the instrument’s access door.

Warning: The ACQUITY UPLC system connections, ESI probe, and source can be contaminated with biohazardous and/or toxic materials. Always wear chemical-resistant, powder-free gloves while performing this procedure.

Warning: To avoid electric shock, ensure that the instrument is suitably prepared before commencing this procedure.

Warning: The source can be hot. To avoid burn injuries, take great care while working with the instrument’s access door open.

3-2 ESI and ESCi Modes of Operation

3. Remove the protective sleeve, if fitted, from the ESI probe tip.

4. Ensure that the contacts on the ESI probe align with the probe adjuster assembly contacts, and carefully slide the ESI probe into the hole in the probe adjuster assembly.

5. Secure the ESI probe by tightening the two thumbscrews.

See also: Figure titled “ESI probe mounted on the source enclosure, showing the connections to the front panel” on page 3-4.

Warning: The ESI probe tip is sharp. To avoid puncture wounds, handle the ESI probe with care.

TP02632

ESI probe

Probe adjuster assembly contacts

Probe adjuster assembly

Installing the ESI probe 3-3

ESI probe mounted on the source enclosure, showing the connections to the front panel:

6. Connect the ESI probe’s PTFE tube to the nebulizer gas connection.

7. Ensure that the probe adjuster assembly’s electrical lead is connected to the instrument’s probe connection.

8. Connect the ESI probe’s electrical lead to the instrument’s HV connection.

9. Connect the diverter valve to the ESI probe; use tubing of the appropriate internal diameter (ID).

Tip: Two tubes of different ID are supplied with the instrument.

Requirement: If you are replacing the tubing supplied with the instrument, you must minimize the length of the tube connecting the diverter valve to the ESI probe. This minimizes delays and dispersion.

10. Close the instrument’s access door.

Warning: To avoid electric shock, do not use stainless steel tubing to connect the diverter valve to the ESI probe; use the PEEK™ tubing supplied with the instrument.

Thumbscrew

ESI probe

Nebulizer gas connection

Desolvation gas connection

Probe adjuster assembly

ESI probe electrical lead

Probe adjuster assembly electrical lead

Vernier probe adjuster

Diverter valve


Installing the corona pin

Required materials• Chemical-resistant, powder-free gloves

• Needle-nose pliers

• 80:20 acetonitrile/water

To install the corona pin:

1. In the ACQUITY UPLC Console, click Standby , and confirm that the Operate indicator is not illuminated.


3. Unlatch the source enclosure door’s handle and open the door.


Warning: To avoid electric shock, ensure that the instrument is in Standby mode when commencing this procedure.


Warning: The probe tip is sharp. To avoid puncture wounds, take great care while working with the source enclosure door open if an ESI probe is fitted.

Caution: Do not apply any downward force to the source enclosure door while the door is open.

Installing the corona pin 3-5

4. Use the needle-nose pliers to remove the blanking plug from the corona pin mounting contact.

Tip: Store the blanking plug in a safe location.

Corona pin mounting contact:

5. Use the needle-nose pliers to fit the corona pin to the mounting contact.

Requirement: Ensure that the corona pin is securely mounted and that its tip aligns with the sample cone orifice.

Warning: The corona pin tip is sharp. To avoid puncture wounds, handle the corona pin with care.

Caution: To avoid damaging to the corona pin’s tip and bending the pin, use the needle nose pliers to grip the corona pin at the end that fits into the mounting contact.

TP02660

Corona pin mounting contact blanking plug


Corona pin:

6. Use the vernier probe adjuster to position the ESI probe tip so that it is pointing approximately mid-way between the tips of the sample cone and corona pin.

See also: Figure titled “ESI probe mounted on the source enclosure, showing the connections to the front panel” on page 3-4.

7. Close the source enclosure door and fasten the handle.


Optimizing the ESI probe for ESCi operation

Required materials80:20 acetonitrile/water

To optimize the ESI probe for ESCi operation:

1. In the MassLynx window, click MS Tune.

2. In the Tune window, click Setup > Inter-scan Setup.

3. In the Inter-scan Setup dialog box, click Reset to Defaults.

TP02695

Corona pin

Sample cone tip

ESI probe tip

Optimizing the ESI probe for ESCi operation 3-7

4. Click OK.

5. In the MassLynx Tune window, click Ion Mode > ESCi+.

6. Select box numbers 1 and 2, clear box numbers 3 and 4 (above the peak display).

7. In row 1, set Ion Mode to ES.

8. In row 2, set Ion Mode to APcI.

9. In each row, set Mass to 42 and Span to 5.

10. Start an infusion of 80:20 acetonitrile/water.

11. Use the vernier probe adjuster to ensure that the ESI probe tip is pointing approximately mid-way between the tips of the sample cone and corona pin.

12. On the Tune window, observe the 42 Da peak in the ES+ and APcI+ peak displays, and increase the values of Capillary (kV) and Corona [(µA) in the current mode or kV in the voltage mode] to produce the most intense ESI+ and APcI+ signal.

13. Use the vernier probe adjuster to gradually move the probe bi-directionally to determine the best position for both the ESI+ and APcI+ signals.

Tip: You can find a position between the two modes that yields a relatively optimized signal.

14. To determine whether you have discrete ionization in the ESI or APcI mode, set the Capillary parameter to 0 kV and observe that little or no signal remains in ESI mode. Then set the Corona parameter to 0 µA or 0 kV, and observe that little or no signal remains in APcI mode.

Result: The ESI probe is now optimized for ESCi mode.

Tip: If necessary, repeat the above procedure using the analyte of interest, since ionization potentials may vary with different samples.


Removing the corona pin

Required materials• Chemical-resistant, powder-free gloves

• Needle-nose pliers

To remove the corona pin:

1. In the ACQUITY UPLC Console, click Standby , and confirm that the Operate indicator is not illuminated.


3. Unlatch the source enclosure door’s handle and open the door.

Warning: The ACQUITY UPLC system connections, corona pin, ESI probe, and source can be contaminated with biohazardous and/or toxic materials. Always wear chemical-resistant, powder-free gloves while performing this procedure.

Warning: To avoid electric shock, ensure that the instrument is in Standby mode when commencing this procedure.


Warning: The probe tip is sharp. To avoid puncture wounds, take great care while working with the source enclosure door open if an ESI probe is fitted.

Caution: Do not apply any downward force to the source enclosure door while the door is open.

Removing the corona pin 3-9

4. Use the needle-nose pliers to remove the corona pin from its mounting contact.

Tip: Store the corona pin in a safe location.

See also: Figure titled “Corona pin:” on page 3-7.

5. Use the needle-nose pliers to fit the blanking plug to the corona pin mounting contact.

See also: Figure titled “Corona pin mounting contact:” on page 3-6.

6. Close the source enclosure door and fasten the handle.


Warning: The corona pin tip is sharp. To avoid puncture wounds, handle the corona pin with care.

Caution: To avoid damaging to the corona pin’s tip and bending the pin, use the needle nose pliers to grip the corona pin at the end that fits into the mounting contact.


Removing the ESI probe

Required materialsChemical-resistant, powder-free gloves

To remove the ESI probe:

1. Prepare the instrument for working on the source.

See also: Chapter 5 of the Waters TQ Detector Operator’s Guide.


3. Disconnect the diverter valve tubing from the ESI probe.

4. Disconnect the ESI probe’s electrical lead from the high voltage connection.

5. Disconnect the ESI probe’s PTFE tube from the nebulizer gas connection.

6. Undo the two thumbscrews securing the ESI probe to the probe adjuster assembly.

7. Carefully remove the ESI probe from the probe adjuster assembly.


Warning: To avoid electric shock, ensure that the instrument is suitably prepared before commencing this procedure.

Warning: The ESI probe and source can be hot. To avoid burn injuries, take great care while working with the instrument’s access door open.

Warning: The ESI probe tip is sharp. To avoid puncture wounds, handle the probe with care.

Removing the ESI probe 3-11

8. If available, fit the protective sleeve to the ESI probe tip.



4 Operating the Waters TQ Detector

This chapter is an introduction to operating your TQ Detector; it explains these tasks:

• Setting-up your TQ Detector.

• Performing a sample tune.

• Developing instrument methods.

• Verifying the system.

Contents:

Topic Page

Setting-up the instrument 4-2

Performing a sample tune 4-7

Developing experiment methods 4-9

Verifying the system using System QC 4-11

4-1

Setting-up the instrument

The IntelliStart instrument setup calibrates the instrument and then, by default, performs a sample tune. If calibration is unnecessary, you can perform only a sample tune.

See also: “Performing a sample tune” on page 4-7.

Tip: Instrument setup need only be performed every three to six months, depending on your usage requirements.

In the following example, sodium cesium iodide is used as the calibrant solution and sulfadimethoxine the tune sample.

Tip: You can substitute solutions suitable for your requirements.


Required materials• Sodium cesium iodide solution (2 ng/µL)

• Sulfadimethoxine solution (100 pg/µL)

To prepare the IntelliStart fluidics system:

See also: “IntelliStart fluidics system operation” on page 1-17.

1. Ensure that IntelliStart fluidics system’s reservoir A is filled with sodium cesium iodide solution.

2. Ensure that reservoir B is filled with sulfadimethoxine solution.

Requirement: Ensure that there is enough solution in each reservoir for approximately 5 minutes of operation during the set-up procedure.

Recommendation: In general, place calibrant solution in reservoir A and tune sample in reservoir B.

3. In the MassLynx window, click MS Console.



6. In the ACQUITY UPLC Console window, click the current flow rate.

7. In the Select Flow Rate dialog box, enter a 50 µL/min flow rate.

4-2 Operating the Waters TQ Detector

8. Click OK.

9. Click to purge the system.

Tip: System purge takes approximately 2 minutes.

10. Repeat step 9.

Rationale: Purging the IntelliStart fluidics system twice ensures that any traces of previously used solutions are removed from the system.

11. Click on the currently-selected reservoir display.

12. In the Select Reservoir dialog box, select the reservoir not currently selected.

13. Click OK.

Result: The other reservoir is selected, and the system is purged.

14. When the purge finishes, click to purge the system again.

To specify the instrument set-up parameters:


2. Ensure that Ion Mode is ES+.

3. In the ACQUITY UPLC Console system tree, click IntelliStart.

4. Ensure that Instrument Setup is selected.

5. If you require system pre-checking, click Pre-checks.

Rationale: If Pre-checks is selected, when IntelliStart starts the instrument set-up, it determines whether the existing calibration is still valid. If so, it does not perform a full calibration but proceeds to sample tuning the instrument.

See also: TQ Detector online Help topic “IntelliStart flow diagram”.

6. Click Start to open the IntelliStart Setup Parameters dialog box.

7. In the Instrument Setup tab’s Reference drop-down list, click Naics.

Rationale: Naics is the calibration reference file for sodium cesium iodide when working in ES+ ion mode.


8. Click Fill from reference file.

Result: The default mass values appear in the Instrument Tune Masses text boxes.

Rule: You must click Fill from reference file every time you select a new calibration reference file.

Instrument Setup Parameters dialog box Instrument Setup tab with default parameters:

Tip: You can use alternative reference solutions to calibrate at higher masses.

9. The tune and calibration results are written to the files in the Instrument Tune and Calibration text boxes; you can use the default files, enter new names, or Browse for files.


To specify the sample tune parameters:

1. In the IntelliStart Setup Parameters dialog box, click the Sample Tune tab.

2. Click Copy Instrument Setup masses.

Rationale: The Tune Masses from the Instrument Setup tab are copied into the Sample Tune Masses boxes.

Instrument Setup Parameters dialog box Sample Tune tab:

3. In the Tune text box, enter sulfadimethoxine.ipr.

Rationale: The sample tune results are written to this file.


Instrument Setup Parameters dialog box Sample Tune tab with example sample tune parameters:

To start instrument set-up:

1. Click Start .

Result: A message appears reminding you to ensure that the calibrant solutions, calibration parameters and LC flow are set correctly.

2. Click OK.

Result: Instrument set-up starts. An autotune on the calibrant is followed by automatic calibration. The ACQUITY UPLC Console displays the progress of the setup.


Example ACQUITY UPLC Console display during calibration:

IntelliStart creates tune and calibration files, which are saved as specified on the Instrument Setup Parameters dialog box’s Instrument Setup tab. Once calibration is complete, the sample tune starts on the four masses defined in the IntelliStart Setup Parameters dialog box. When the sample tune is complete, the results are written to the Sulfadimethoxine.ipr file.

Performing a sample tune

In the following example, sulfadimethoxine is used as the tuning sample.

Required materialsSulfadimethoxine solution (100 pg/µL)

To perform a sample tune:

1. Prepare the IntelliStart fluidics system with sulfadimethoxine solution in reservoir B.

See also: “To prepare the IntelliStart fluidics system:” on page 4-2.

Performing a sample tune 4-7


3. Ensure that the Ion Mode is ES+.


5. Clear the Instrument Setup check box.


Rationale: If Pre-checks is selected, when IntelliStart starts the sample tune, it determines whether the existing tune is still valid. If so, no sample tune is performed.


7. Click Start .

8. In the IntelliStart Setup Parameters dialog box’s Sample Tune tab, specify the sample tune parameters as described in “To specify the sample tune parameters:” on page 4-5.

9. Click Start .

Result: A message appears reminding you to ensure that the tune solutions, tune parameters, and LC flow are set correctly.

10. Click OK.

Result: Sample tune starts. The ACQUITY UPLC Console displays the progress of the tune. When the sample tune is complete, the results are written to the Sulfadimethoxine.ipr file.


Developing experiment methods

IntelliStart allows you to automatically develop quantitative MRM or SIR methods for compounds of interest. Up to four compounds can be handled in a single process. Up to three transitions can be selected for each compound.In this example, methods for sulfadimethoxine are created.

Required materialsSulfadimethoxine solution (100 pg/µL)

To create a method:

1. Prepare the IntelliStart fluidics system with sulfadimethoxine solution in reservoir B.

See also: “To prepare the IntelliStart fluidics system:” on page 4-2.


3. Ensure that the Ion Mode is ES+.


5. Clear the Instrument Setup check box.

6. Click Develop Method.



8. Click Start .

9. In the IntelliStart Setup Parameters dialog box, click the Method Developer tab.

Note: The masses, tune file and IntelliStart fluidics system parameters already set in IntelliStart are used.

10. Select the parameters shown in the following figure.

Developing experiment methods 4-9

Instrument Setup Parameters dialog box Method Developer tab:

In this case, Sulfadimethoxine.exp is the method file created.

The Validation pane selections save optimization data for validation purposes and creates an autotune report file (Sulfadimethoxine.xml).

11. Click Start .

Result: A message appears reminding you to ensure that the sample solutions, sample tune parameters and LC flow are set correctly.

12. Click OK.

Result: Method development starts. The ACQUITY UPLC Console displays the progress of the method development. When the method development is complete, the method is written to the Sulfadimethoxine.exp file.

The Validation pane selection saves optimization data for validation purposes, and creates an autotune report file (Sulfadimethoxine.xml).

A green check mark indicates a successful run; a red cross indicates a failure.


Verifying the system using System QC

The System QC function uses pre-defined LC/MS methods to test the system by injecting a diagnostic sample on-column. The sample helps identify any problems with the configured system by providing these data:

• Sensitivity (signal-to-noise ratio)

• Response (peak area, peak height)

• Chromatographic performance (peak width, retention time)

System QC can be activated in several ways:

• Manually, from the ACQUITY UPLC Console.

• Manually, as part of an ACQUITY UPLC Console Autotune sequence.

• Automatically, based on a scheduled date and time.

The following example shows you how to perform a manual verification.

See also: The MassLynx online Help for details of how to perform verification manually, as part of an Autotune sequence or automatically based on a scheduled date and time.

Multiple injections are processed using the OpenLynx Application-Manager. Measurements are reported for each chosen parameter across the repeat analyses. If results are within user-defined tolerances, the LC/MS system is ready to use.

The System Ready results are logged via the ACQUITY UPLC Console but can also appear in a printed report. The raw data and OpenLynx browser report are also stored for records.

Requirement: For System QC to work, OpenLynx must be installed with MassLynx. OpenLynx is normally installed by default. If it has not been installed, run the installation DVD, select Modify, and select OpenLynx when prompted.

Before running System QC you need a suitable tune file, method file and inlet file. You can use IntelliStart to create a tune file and develop a method for the sample you want to use for System QC.

See also: “Performing a sample tune” on page 4-7 and “Developing experiment methods” on page 4-9.


An inlet file must be created independently.

See also: The MassLynx online Help.

Requirement: IntelliStart reads the tune, inlet, and method files from the System QC project (SystemQC.pro). You must save the tune, inlet, and method files to the Aqudb folder of the System QC project before performing the verification.

To perform a manual verification using System QC:

1. In the ACQUITY UPLC Console, click System QC.

2. Click Start .

Instrument Setup Parameters dialog box System QC tab with default parameters:


3. In the IntelliStart Setup Parameters dialog box’s System QC tab, enter the number of pre-run and run injections, the vial (that is, the position of your sample in the Autosampler) and the injection volume.

Tip: At least two injections are required for a run, because some of the validation parameters use standard deviation. A pre-run sample is used to clean and stabilize the system. You need not perform a pre-run prior to a run.

Recommendation: Perform at least three injections.

4. Enter a tune, LC and inlet file, or use the browse button to select a file.

5. Select whether you are using a Scanning or SIR/MRM method. If using a scanning method, enter a Mass on which to perform peak detection.

6. Select Print Report to print a report of the results.

7. Select the Acceptance parameters.

Instrument Setup Parameters dialog box System QC tab Acceptance parameters :

Parameter Description

Retention Time Enter the expected retention time in the Set Point box and enter the tolerance. If, for any sample, the peak top is outside the range, the test reflects a failure.

Peak Area The variation in peak area is calculated by percentage Relative Standard Deviation (%RSD).%RSD = (Standard Deviation/Average Area) × 100%Enter a tolerance in %RSD, above which the test reflects a failure.

Peak Height This is similar to Peak Area but using Peak Height instead.

Peak Width Enter the expected peak width in the Set Point box and enter the tolerance. If, for any sample, the peak width falls outside the range, the test reflects a failure.


Tip: The calculations do not include injections where no peak was found. For example, if only eight out of ten injections evidence detected peaks, the acceptance test results would be based on eight values rather than ten. However, the System QC process would always be marked as “failed” due to the missing peaks.

8. Click Start .

Result: A message appears reminding you to ensure that the sample vial, QC parameters and LC flow are set correctly.

9. Click OK.

Result: System QC starts. The ACQUITY UPLC Console displays its progress.

Signal/Noise In Signal/Noise enter the signal-to-noise ratio below which the test reflects a failure.In Noise Range, enter the range in retention time window over which the noise should be calculated.Select how the noise is calculated, either RMS or Peak-to-Peak.Tip: The signal is the peak height.

Instrument Setup Parameters dialog box System QC tab Acceptance parameters (Continued):

Parameter Description


A 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. This appendix presents all the safety symbols and statements that apply to the entire line of Waters products.

Contents:

Topic Page

Warning symbols A-2

Caution symbol A-5

Warnings that apply to all Waters instruments A-5

Electrical and handling symbols A-13

A-1

Warning symbols

Warning symbols alert you to the risk of death, injury, or seriously adverse physiological reactions associated with an instrument’s use or misuse. 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.

Task-specific hazard warningsThe following warning symbols alert you to risks that can arise when you operate or maintain an instrument or instrument component. Such risks include burn injuries, electric shocks, ultraviolet radiation exposures, and others.

When the following symbols appear in a manual’s narratives or procedures, their accompanying text identifies the specific risk and explains how to avoid it.

Warning: (General risk of danger. When this symbol appears on an instrument, consult the instrument’s user documentation for important safety-related information before you use the instrument.)

Warning: (Risk of burn injury from contacting hot surfaces.)

Warning: (Risk of electric shock.)

Warning: (Risk of fire)

Warning: (Risk of needle puncture.)

Warning: (Risk of injury caused by moving machinery.)

Warning: (Risk of exposure to ultraviolet radiation.)

Warning: (Risk of contacting corrosive substances.)

Warning: (Risk of exposure to a toxic substance.)

A-2 Safety Advisories

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.

Mass spectrometer flammable solvents warning

This warning applies to instruments operated with flammable solvents.

Warning: (Risk of personal exposure to laser radiation.)

Warning: (Risk of exposure to biological agents that can pose a serious health threat.)

Warning: Pressurized nonmetallic, or polymer, tubing can burst. Observe these precautions when working around such tubing:• Wear eye protection.• Extinguish all nearby flames.• Do not use tubing that is, or has been, stressed or kinked.• Do not expose nonmetallic tubing to incompatible compounds like

tetrahydrofuran (THF) and nitric or sulfuric acids.• Be aware that some compounds, like methylene chloride and

dimethyl sulfoxide, can cause nonmetallic tubing to swell, which significantly reduces the pressure at which the tubing can rupture.

Warning: Where significant quantities of flammable solvents are involved, a continuous flow of nitrogen into the ion source is required to prevent possible ignition in that enclosed space. Ensure that the nitrogen supply pressure never falls below 400 kPa (4 bar, 58 psi) during an analysis in which flammable solvents are used. Also ensure a gas-fail connection is connected to the HPLC system so that the LC solvent flow stops if the nitrogen supply fails.

Warning symbols A-3

Mass spectrometer shock hazard

This warning applies to all Waters mass spectrometers.

This warning applies to certain instruments when they are in Operate mode.

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.

Warning: To avoid electric shock, do not remove the mass spectrometer’s protective panels. The components they cover are not user-serviceable.

Warning: High voltages can be present at certain external surfaces of the mass spectrometer when the instrument is in Operate mode. To avoid non-lethal electric shock, make sure the instrument is in Standby mode before touching areas marked with this high voltage warning symbol.

Warning: Waters's instruments and software can be used to analyze or process potentially infectious human-sourced products, inactivated microorganisms, and other biological materials. To avoid infection with these agents, assume that all biological fluids are infectious, observe good laboratory practices and, consult your organization’s biohazard safety representative regarding their proper use and handling. Specific precautions appear in the latest edition of the US National Institutes of Health (NIH) publication, Biosafety in Microbiological and Biomedical Laboratories (BMBL).


Chemical hazard warning

This warning applies to Waters instruments that can process corrosive, toxic, flammable, or other types of hazardous material.

Caution symbol

The caution symbol signifies that an instrument’s use or misuse can damage the instrument or compromise a sample’s integrity. The following symbol and its associated statement are typical of the kind that alert you to the risk of damaging the instrument or sample.

Warnings that apply to all Waters instruments

When operating this device, follow standard quality control procedures and the the equipment guidelines in this section.

Warning: Waters instruments can be used to analyze or 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 representative regarding proper use and handling. Guidelines are provided in the latest edition of the National Research Council's publication, Prudent Practices in the Laboratory: Handling and Disposal of Chemicals.

Caution: To avoid damage, do not use abrasives or solvents to clean the instrument’s case.

Caution symbol A-5

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 exploiter l’équipement.

Achtung: Jedwede Änderungen oder Modifikationen an dem Gerät ohne die ausdrückliche Genehmigung der für die ordnungsgemäße Funktionstüchtigkeit verantwortlichen Personen kann zum Entzug der Bedienungsbefugnis des Systems führen.

Avvertenza: eventuali modifiche o alterazioni apportate a questa unità e non espressamente approvate da un ente responsabile per la conformità annulleranno l’autorità dell’utente ad operare l’apparecchiatura.

Atencion: cualquier cambio o modificación efectuado en esta unidad que no haya sido expresamente aprobado por la parte responsable del cumplimiento puede anular la autorización del usuario para utilizar el equipo.


Warning: Use caution when working with any polymer tubing under pressure:• Always wear eye protection when near pressurized polymer tubing.• Extinguish all nearby flames.• Do not use tubing that has been severely stressed or kinked.• Do not use nonmetallic tubing with tetrahydrofuran (THF) or concentrated

nitric or sulfuric acids.• Be aware that methylene chloride and dimethyl sulfoxide cause

nonmetallic tubing to swell, which greatly reduces the rupture pressure of the tubing.

Attention: Manipulez les tubes en polymère sous pression avec precaution:• Portez systématiquement des lunettes de protection lorsque vous vous

trouvez à proximité de tubes en polymère pressurisés.• Eteignez toute flamme se trouvant à proximité de l’instrument.• Evitez d'utiliser des tubes sévèrement déformés ou endommagés.• Evitez d'utiliser des tubes non métalliques avec du tétrahydrofurane

(THF) ou de l'acide sulfurique ou nitrique concentré.• Sachez que le chlorure de méthylène et le diméthylesulfoxyde entraînent le

gonflement des tuyaux non métalliques, ce qui réduit considérablement leur pression de rupture.

Vorsicht: Bei der Arbeit mit Polymerschläuchen unter Druck ist besondere Vorsicht angebracht:• In der Nähe von unter Druck stehenden Polymerschläuchen stets

Schutzbrille tragen.• Alle offenen Flammen in der Nähe löschen.• 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.


Attenzione: prestare attenzione durante l’utilizzo dei tubi di polimero pressurizzati:• Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero

pressurizzati.• Estinguere ogni fonte di ignizione circostante.• Non utilizzare tubi soggetti che hanno subito sollecitazioni eccessive o son

stati incurvati.• Non utilizzare tubi non metallici con tetraidrofurano (THF) o acido

solforico o nitrico concentrato.• Tenere presente che il cloruro di metilene e il dimetilsolfossido provocano

rigonfiamento nei tubi non metallici, riducendo notevolmente la resistenza alla rottura dei tubi stessi.

Advertencia: se recomienda precaución cuando se trabaje con tubos de polímero sometidos a presión:• El usuario deberá protegerse siempre los ojos cuando trabaje cerca de

tubos de polímero sometidos a presión.• Si hubiera alguna llama las proximidades.• No se debe trabajar con tubos que se hayan doblado o sometido a altas

presiones.• Es necesario utilizar tubos de metal cuando se trabaje con

tetrahidrofurano (THF) o ácidos nítrico o sulfúrico concentrados.• Hay que tener en cuenta que el cloruro de metileno y el sulfóxido de

dimetilo dilatan los tubos no metálicos, lo que reduce la presión de ruptura de los tubos.



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.

Vorsicht: Der Benutzer wird darauf aufmerksam gemacht, dass bei unsachgemäßer Verwenddung des Gerätes unter Umständen nicht ordnungsgemäß funktionieren.

Attenzione: l’utente deve essere al corrente del fatto che, se l’apparecchiatura viene usta in un modo specificato dal produttore, la protezione fornita dall’apparecchiatura potrà essere invalidata.

Advertencia: el usuario deberá saber que si el equipo se utiliza de forma distinta a la especificada por el fabricante, las medidas de protección del equipo podrían ser insuficientes.


Warning: To protect against fire hazard, replace fuses with those of the same type and rating.

Attention: Remplacez toujours les fusibles par d’autres du même type et de la même puissance afin d’éviter tout risque d’incendie.

Vorsicht: Zum Schutz gegen Feuergefahr die Sicherungen nur mit Sicherungen des gleichen Typs und Nennwertes ersetzen.

Attenzione: per una buona protezione contro i rischi di incendio, sostituire i fusibili con altri dello stesso tipo e amperaggio.

Advertencia: sustituya los fusibles por otros del mismo tipo y características para evitar el riesgo de incendio.


Warning: To avoid possible electrical shock, disconnect the power cord before servicing the instrument.

Attention: Afin d’éviter toute possibilité de commotion électrique, débranchez le cordon d’alimentation de la prise avant d’effectuer la maintenance de l’instrument.

Vorsicht: Zur Vermeidung von Stromschlägen sollte das Gerät vor der Wartung vom Netz getrennt werden.

Attenzione: per evitare il rischio di scossa elettrica, scollegare il cavo di alimentazione prima di svolgere la manutenzione dello strumento.

Precaución: para evitar descargas eléctricas, desenchufe el cable de alimentación del instrumento antes de realizar cualquier reparación.


Electrical and handling symbols

Electrical symbolsThese 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.

Electrical and handling symbols A-13

Handling symbolsThese 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!


Documents

Waters TQ Detector Operator’s Guide - Xi'an University ...jg.xauat.edu.cn/labs/hjgch/files/pdf/2014/20141216105217_9573.pdf · 1 Waters TQ Detector Overview This chapter describes