LocksprayTM

User's Guide

Micromass UK Limited

Floats RoadWythenshawe

M23 9LZTel: +44 161 945 4170 Fax: +44 161 998 8915

Tudor RoadAltrinchamWA14 5RZ

Tel: +44 161 282 9666 Fax: +44 161 282 4400

http://www.micromass.co.uk

The instrument is marked with this symbol where high voltages arepresent.

The instrument is marked with this symbol where hot surfaces arepresent.

The instrument is marked with this symbol where the user should refer tothis User's Guide for instructions which may prevent damage to the

instrument.

Warnings are given throughout this manual where care is required to avoid personalinjury.

If the instrument is used in a manner not specified by the manufacturer, the protectionprovided by the equipment may be impaired.

All information contained in this manual is believed to be correct at the time ofpublication. The publishers and their agents shall not be liable for errors contained

herein nor for incidental or consequential damages in connection with the furnishing,performance or use of this material. All product specifications, as well as theinformation contained in this manual, are subject to change without notice.

Micromass ® is a registered trade mark of Micromass Limited(Reg. U.S. Pat. & T.M. Off.).

Code Number 6666531 Issue 1© Micromass Ltd.

Table of Contents

LocksprayTM

User's Guide

ContentsDescription

Introduction 5

Installation7

Operation and Tuning9

Data Processing11

Maintenance and Fault FindingOscillating Baffle 13Reference Probe Capillary 14Reference Probe Tips 15Possible Faults 15

Appendix A - LockSpray Control UnitElectrical Connections 17Cleaning 18

Appendix B - Technical SpecificationTechnical Specification 19

Appendix C - Spare Parts ListSpare Parts List 21

Table of Contents

LocksprayTM

User's Guide

Table of Contents

LocksprayTM

User's Guide

DescriptionIntroduction

The LockSpray interface facilitates acquisition of exact mass data on electrospraytime-of-flight instruments using lock mass correction whilst avoiding many of theproblems associated with post-column teeing in of a reference material.

Current practice for obtaining exact mass data from a time of flight instrument is toadd a reference compound (lock mass) to the LC eluent prior to entry into the massspectrometer which can then be used to correct for drifts in the mass scale. There areinherent problems with this approach which can lead to poor exact mass capability:suppression of the analyte by the reference or vice versa; mass interferences betweenthe analyte and reference; clustering of the analyte and reference, and variation in thereference signal strength with changes in LC gradient. In addition, the process ofteeing in the reference can in some cases affect chromatographic performance.

DescriptionPage 5

LockSprayTM

User's Guide

LockSpray provides the capability to introduce the reference flow through a differentelectrospray probe from that of the analyte and using an oscillating baffle allowsanalysis of each spray independently, thus avoiding the problems outlined above. Thediagram below shows schematically how this is achieved.

The LockSpray source mounts directly onto the standard z-spray interface andprovides a low flow capability electrospray probe (<20 µl/min) orthogonal to thestandard electrospray probe for introduction of the reference compound. This probehas the same electrospray voltage applied as the standard probe and the nebulising gaslines are coupled. Heated desolvation gas for the reference comes indirectly from thestandard desolvation heater. Also on the LockSpray source is a baffle coupled to aposition indexed stepper motor which admits the spray from either the standardanalyte electrospray probe or the reference probe to the sampling cone for acquisition.Spray indexing allows the analyte and reference data to be acquired into separate datafiles and the design of the baffle produces negligible cross talk between the twosprays. Data from the reference spray are used to calculate a correction factor for themass scale which is then applied to the analyte data to provide exact massinformation.

DescriptionPage 6

LockSprayTM

User's Guide

AnalyteInlet

DESOLVATIONGAS

SamplingCone

IonBlock Hexapole

Ion Bridge

Baffle

ReferenceInlet

Installation

Remove the standard probe, probe adjustment flange, glass cover and mountingpillars. Leave the standard baffle in place and ensure it is pushed fully into itssupport.

Using the mounting pillars supplied, attach the LockSpray source to the Z-sprayinterface, taking great care not to damage the sampling cone with the baffle.Ensure that the pillars are fully tightened using a small bar inserted into theholes to act as a lever.

Place the new glass cover on the mounting pillars and prior to re-attaching theprobe adjustment flange, ensure that the in/out adjustment is fully out. Re-insertthe standard electrospray probe.

Connect the desolvation gas heater cable to the Probes socket on the instrumentfront panel and the PTFE gas line to the Desolvation Gas connector.

InstallationPage 7

LockSprayTM

User's Guide

Stepper Motor Cover

Baffle

Reference Probe

Glass Housing

Support Pillars

Standard Electrospray Probe

Connect the high voltage cable from the standard electrospray probe into thesocket on the reference probe, then connect the high voltage cable from thereference probe to the Capillary / Corona connector on the instrument frontpanel.

Using the legris Tee piece provided couple the PTFE gas lines from the twoprobes to the Nebuliser Gas connector.

Connect the cable from the LockSpray control unit to the D-Type socket on theunderside of the motor cable. Refer to Appendix A for instructions on installingthe LockSpray control box.

In the main MassLynx menu select Tools.

From the Options dialogue check the LockSpray box.

InstallationPage 8

LockSprayTM

User's Guide

Operation and TuningThe procedures for undertaking exact mass work using lock mass correction onthe time-of-flight instruments are described in the User’s Guide and should beused with the LockSpray source.

The standard electrospray probe can be operated as on a standard electrospraysource, coupled to an appropriate LC system. The reference sprayer should becoupled to a pump capable of delivering the reference solution in a pulse-freemanner over typically 5–20 µl/min, a syringe pump is particularly suitable. Thefrequency with which the reference spray is sampled is user configurable butshould generally be set in the 2 to 10 second range. The acquisition time on thereference spray is the same as set for the analyte spray.

The operating conditions for the standard electrospray probe in LockSpray areessentially the same as for the standard electrospray source and the user shouldrefer to the appropriate sections of the instrument User’s Guide for optimisation.The positioning of the probe tip with respect to the sample cone and theoptimum capillary voltage may differ somewhat from the standard arrangementdue to the proximity of the earthed baffle. Care should be taken when settingthese parameters to ensure that the probe does not electrically discharge to thebaffle.

There are no positional adjustments possible for the reference probe. To obtainthe required reference signal for exact mass work, the concentration and flowrate of the reference material should be adjusted accordingly once the standardsprayer has been optimised for the analyte. The cone voltage for use with thereference compound can also be optimised as it is configurable in the experimentset-up from the sample list. The signals from the analyte or reference sprayercan be observed for tuning purposes by using the LockSpray selection menu onthe ES Source window of the tune page.

Operation and TuningPage 9

LockSprayTM

User's Guide

Operation and TuningPage 10

LockSprayTM

User's Guide

Data ProcessingDuring the course of an acquisition the analyte and reference data from the LockSpraysource are stored in the same folder as two separate functions and can be viewedindependently.

When data processing with LockSpray the number of reference scans to average needsto be set. This parameter determines how many data points of the reference file up tothe time of the combined range of analyte data will be averaged to provide acorrection factor. Values between 5 and 10 are usually suitable. If the value used istoo low then ion statistics can affect the mass accuracy, too high and there is thepossibility that the instrument will have drifted slightly giving rise to an inappropriatecorrection factor.

Data ProcessingPage 11

LockSprayTM

User's Guide

Data ProcessingPage 12

LockSprayTM

User's Guide

Maintenance and Fault FindingGeneral maintenance and fault finding for a Z-spray source operating in theelectrospray mode can be found in the appropriate section of the User’s Guide.

Oscillating BaffleTo remove the baffle for cleaning,

undo the single screw holding it to the shaft. In order to gain access to the screwwith the LockSpray source still mounted on the instrument, the motor cable willneed to be unplugged to allow free rotation of the baffle.

Put the baffle in a beaker of 50:50 acetonitrile:water or 50:50 methanol waterwith 1% formic acid and place in an ultrasonic bath for 15 minutes. Rinse withwater after cleaning.

When replacing the baffle, ensure it locates correctly on the square end of theshaft and check that it is oriented such that the reference and analyte sprays areselected correctly from the tune page.

Maintenance and Fault FindingPage 13

LockSprayTM

User's Guide

Reference Probe Capillary

If the stainless steel reference capillary cannot be cleared, or if it is contaminated ordamaged, replace it as follows:

Ensure that the system is out of operate.

Disconnect the reference line from the probe and remove the finger-tight nut.

Disconnect the HV cable from the reference probe at the instrument front panel.

Disconnect the HV cable from the analyte probe at the reference probeconnector.

Maintenance and Fault FindingPage 14

LockSprayTM

User's Guide

ReferenceCapillary

Tube

ConductiveSleeve

PTFE LinerTube

(20mm)

PTFE LinerTube

(50mm)

GrubScrew

LC Union

O Ring

Probe Tip

AdapterNut

Disconnect the reference probe nebulising gas tube.

Undo the two bolts holding the reference probe to the LockSpray housing andcarefully withdraw the probe.

Loosen the grub screw retaining the LC union.

Remove the probe cover by unscrewing the three retaining screws.

Unscrew and remove the probe tip.

Remove the LC union, adapter nut and conductive sleeve. Withdraw and discardthe stainless steel reference capillary.

Sleeve one end of a new reference capillary with the PTFE liner tube.

Using a GVF/16 ferrule and the adapter nut, connect the sample capillary to theLC union, ensuring that both the liner tube and sample capillary are fully buttedinto the LC union.

Slide the conductive sleeve onto the reference capillary, feed the capillarythrough the probe and tighten down the adapter nut.

Replace the probe tip and adjust so that 0.5mm of reference capillary protrudesfrom the probe tip.

Replace the probe cover and tighten the grub screw to clamp the LC union.

Re-insert the reference probe with the HV socket in the correct orientation,tighten down and re-connect the gas and HV lines.

Reference Probe TipsTo replace a damaged or blocked probe tip requires the probe to be removed from theLockSpray housing, refer to the section above for replacement of a reference capillary.

Possible FaultsIf the tune page times out when attempting to find either the analyte or referencespray, check that the motor cable is connected and that the LockSpray control unit isswitched on.

If the signals from either spray are unstable or the signal changes when operating, it ispossible that the baffle may need cleaning (see above) or that the probe tips aredischarging to the baffle and so the capillary voltage should be reduced.

Maintenance and Fault FindingPage 15

LockSprayTM

User's Guide

General maintenance and fault finding for a Z-Spray source operating in theelectrospray mode can be found in the appropriate section of the instrumentUser's Guide.

Maintenance and Fault FindingPage 16

LockSprayTM

User's Guide

Appendix A - LockSpray Control UnitCaution: The controller is to be used only as specified in this document, connected asshown to a Micromass mass spectrometer. The controller is designed for indoor useonly. Do not obstruct the air vents (on the underside of the unit).

Electrical ConnectionsWith reference to the diagram below:

Disconnect the SIP cable from the SIP input at the rear of the MS.

Connect the LockSpray signal cable (3892011CC1) to the MS and LockSpray controller.

Connect the SIP cable to the third D connector of the LockSpray signal cable.

Connect the LockSpray motor cable (3892010CC1) to the LockSpray source andcontroller.

Connect the LockSpray Control Unit to the mains supply (there is a 3 pin IECinlet at the rear of the unit). The supply must include a protective ground (Earth).

Switch on the controller via the power switch at the rear of the unit.

Operate the LockSpray system via Micromass’ MassLynx software.

When the LockSpray system is not in use the control unit should be switched off.

Appendix A - LockSpray Control UnitPage 19

LockSprayTM

User's Guide

LockSpraySignal Cable

SIP Cable

LockSprayMotor Cable

Cable ConnectorEMBEDDED PC

Cable Connector SIPCable Connector

MSMS Connector

SIP

To Embedded PC

MassSpectrometer

LockSpray

Cable ConnectorCONTROL UNIT

ControllerConnectorCOMMS

ControllerConnector

MOTOR

Cable ConnectorCONTROL UNIT

LockSprayController

In the unlikely event of a fuse blowing, they are both to be replaced simultaneously asfollows:

Switch off the unit and remove the mains connector at its rear.

Slide out the fuse drawer (built into the IEC inlet).

Replace both fuses with 3.15A (F) quick blow 20mm fuses.

Slide the drawer back and reconnect the mains supply.

Cleaning

Never immerse the unit (partially or fully) in any liquid. Do not use strong solvents toclean the unit. A cloth lightly dampened with soapy water may be used to clean theoutside of the controller.

Appendix A - LockSpray Control UnitPage 20

LockSprayTM

User's Guide

Appendix B - Technical SpecificationTechnical Specification

Item Specification Comment

Operating Temperature 10°C to 35°C

Storage Temperature -20°C to +60°C

Power 90VAC to 264VAC, 47 to 63 Hz Single or dual phase

Operating Humidity 30-90%, noncondensing

Storage Humidity 5-95%, noncondensing

AltitudeUp to 2000 metres (operating)

up to 12 000 metres(non-operating)

Input Surge Current (max)17A @ 120V

32A @ 230V

Safety Ground Leakage Current0.7mA @ 120VAC 60Hz

1.4mA @ 230VAC 50Hz

Input Current (max)1.05A @ 120V

0.51A @ 230V

Input Fuses (User Accessible) 250VAC F 3.15A 20mm Both phases are fused

Appendix B - Technical SpecificationPage 21

LockSprayTM

User's Guide

Appendix B - Technical SpecificationPage 22

LockSprayTM

User's Guide

Appendix C - Spare Parts ListSpare Parts List

Item Code

Probe tip assembly M955090AD1

Reference capillary tube M955785AD1

Conductive sleeve M955714AD1

PTFE Tube (20mm long x 0.010” ID) M955090AD1

PTFE Tube (50mm long x 0.015” ID) M955089AD1

Adapter nut M955541AD1

GVF16 ferrules 6070117

Peek union 6367402

Appendix C - Spare Parts ListPage 23

LockSprayTM

User's Guide

Appendix C - Spare Parts ListPage 24

LockSprayTM

User's Guide