Finnigan LC IsoLinksilab/ThermoManuals/LC/115_6140_LC... · 2008. 10. 14. · _ Finnigan LC IsoLink Operating Manual_ a Thermo ELECTRON CORPORATION Safety and EMC Information

Finnigan™ LC IsoLink™

LC-IRMS Interface

Operating Manual

Revision B115 6140

Prelim

inary V

ersion

Technical information contained in this publication is for reference purposes only and is subject to change without notice. Every effort has been made to supply complete and accurate information; however, Thermo Electron assumes no responsibility and will not be liable for any errors, omissions, damage, or loss that might result from any use of this manual or the information contained therein (even if this information is properly followed and problems still arise).

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Printing History: Revision B printed in 8/10/04.

Published by Product Marketing, Thermo Electron Corporation, Bremen, Germany.Copyright© 2004 Thermo Electron Corporation. All rights reserved. Printed in Germany.

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In compliance with international regulations: If this instrument is used in a manner not specified by Thermo Electron Bremen, the protection provided by the instrument could be impaired.

Safety and EMC Information

In accordance with our commitment to customer service and safety, these instruments have satisfied the requirements for the European CE Mark including the Low Voltage Directive.

Designed, manufactured and tested in an ISO9001 registered facility, this instrument has been shipped to you from our manufacturing facility in a safe condition.

Identifying Safety InformationThis reference manual contains precautionary statements that can prevent personal injury, instrument damage, and loss of data if properly followed. All statements of this nature are called to your attention through the use of bold type and the following icons:

Every instrument has specific hazards, so be sure to read and comply with the following precautions. They will help ensure the safe, long-term use of your system.

1. Before plugging in any of the instrument modules or turning on the power, always make sure that the voltage and fuses are set appropriately for your local line voltage.

2. Only use fuses of the type and current rating specified. Do not use repaired fuses and do not short-circuit the fuse holder.

3. The supplied power cord must be inserted into a power outlet with a protective earth contact (ground). When using an extension cord, make sure that the cord also has an earth contact.

Caution. This instrument must be used as described in this manual. Any use of this instrument in a manner other than described here may result in instrument damage and/or operator injury.

Warning! Cold Surface! Hot Surface Strong Magnetic Field!

High Voltage!

______________________ Finnigan LC IsoLink Operating Manual ____________________ aThermoELECTRON CORPORATION

Safety and EMC InformationIdentifying Safety Information ___________________________________________Finnigan LC IsoLink

4. Do not change the external or internal grounding connections.Tampering with or disconnecting these connections could endanger you and/or damage the system.

5. Never run the system without the housing on. Permanent damage can occur.

6. Do not turn the instrument on if you suspect that it has incurred any kind of electrical damage. Instead, disconnect the power cord and contact a Service Representative for a product evaluation. Do not attempt to use the instrument until it has been evaluated. (Electrical damage may have occurred if the system shows visible signs of damage, or has been transported under severe stress.)

7. Damage can also result if the instrument is stored for prolonged periods under unfavorable conditions (e.g., subjected to heat, water, etc.).

8. Always disconnect the power cord before attempting any type of maintenance.

9. Never try to repair or replace any component of the system that is not described in this manual without the assistance of your service representative.

Caution. The instrument is properly grounded in accordance with regulations when shipped. You do not need to make any changes to the electrical connections or to the instrument’s chassis to ensure safe operation.

Warning. Avoid any contact of the system with liquids! Permanent damage can occur due to high voltage, e.g. leaking liquids might get into contact with electronic components and cause a short circuit.

ThermoELECTRON CORPORATIONb________________________Finnigan LC IsoLink Operating Manual_____________________

ContentsFinnigan LC IsoLink______________________________________________________________________

Contents

Read This First ............................................................................................................................ ix

Changes to the Manual and Online Help ................................................................................................ x

Abbreviations ......................................................................................................................................... xi

Typographical Conventions .................................................................................................................. xvData Input ............................................................................................................................ xvBoxed Information.............................................................................................................. xviTopic Headings.................................................................................................................. xvii

Reply Cards ........................................................................................................................................ xviii

Layout ........................................................................................................................................ 1-1

1.1 Overall View ............................................................................................................................. 1-2

1.2 Front View................................................................................................................................. 1-5

1.3 Functional Separation into Cabinets ......................................................................................... 1-9

1.4 Back View............................................................................................................................... 1-11

1.5 Right View .............................................................................................................................. 1-13

1.6 Left View................................................................................................................................. 1-15

Steps of Analysis ........................................................................................................................ 2-1

2.1 Range of Applications and Analysis Principle ......................................................................... 2-2Range of Applications ........................................................................................................ 2-2Analysis Principle............................................................................................................... 2-2

____________________ Finnigan LC IsoLink Operating Manual _______________________ iThermoELECTRON CORPORATION

Contents_________________________________________________________________Finnigan LC IsoLink

2.2 HPLC Unit................................................................................................................................ 2-6Purging HPLC Pump and HPLC Pulse Damper ................................................................ 2-6Usage of Pure Water........................................................................................................... 2-7

Elimination of Organic Substances ............................................................................. 2-7Water Degassing .......................................................................................................... 2-9Avoidance of Water Regassing .................................................................................. 2-10Keeping Particles away by Eluent Filters .................................................................. 2-10

Backpiston Flushing of HPLC Pump............................................................................... 2-10Operating Additional Devices.......................................................................................... 2-10

Autosampler ...............................................................................................................2-11Oven with HPLC Column ..........................................................................................2-11UV Detector ................................................................................................................2-11In-Line Filter ..............................................................................................................2-11

2.3 Six-Port Valve - Leaving the HPLC Unit and Entering the Interface .................................... 2-12Load Position versus Inject Position................................................................................ 2-12Injection Modes................................................................................................................ 2-13

HPLC Mode ............................................................................................................... 2-13Direct Injection Mode (µ-EA Mode) ......................................................................... 2-13Operating an HPLC Autosampler in Both Modes ..................................................... 2-14

Other Applications of Six-Port Valve .............................................................................. 2-17

2.4 Sample Loop and HPLC Syringe ........................................................................................... 2-18Volume of Sample Loop .................................................................................................. 2-18Overload of Sample Loop ................................................................................................ 2-18Rinsing HPLC Syringe..................................................................................................... 2-18Sucking up Solutions ....................................................................................................... 2-19

2.5 T-Piece .................................................................................................................................... 2-20

2.6 Oxidation Reactor................................................................................................................... 2-21Operating Temperature .................................................................................................... 2-21Avoiding Temperature Fluctuations................................................................................. 2-21

Pulsations of Oxidation Reactor ................................................................................ 2-21Pulsations of HPLC Pump ......................................................................................... 2-22Pulsations of Interface Pumps ................................................................................... 2-22

Tuning JUMO itron 16 Temperature Controller Manually.............................................. 2-23Need of Manual Tuning ............................................................................................. 2-23Parameter Values for Manual Tuning ........................................................................ 2-24Procedure of Manual Tuning ..................................................................................... 2-24

Problem of Incomplete Oxidation.................................................................................... 2-25Cooler............................................................................................................................... 2-25

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2.7 Separation Unit ....................................................................................................................... 2-26Separation Principle.......................................................................................................... 2-26Connections of Separation Unit........................................................................................ 2-26Membrane Damage at Separation Unit ............................................................................ 2-28Leak Check at Separation Unit......................................................................................... 2-28Avoidance of Particle Dispersion ..................................................................................... 2-28Backflush of Separation Unit Capillaries ......................................................................... 2-29

Mechanism of Capillary Plugging ............................................................................. 2-29Step 1 - Capillary Check for Non-Uniform Dropping ............................................... 2-30Step 2 - Backflush Procedure Itself ........................................................................... 2-31Step 3 - After an Unsuccessful Backflush Procedure ................................................ 2-33Step 4 - After a Successful Backflush Procedure ...................................................... 2-33

2.8 Gas Dryer ................................................................................................................................ 2-35Principle of Water Removal ............................................................................................. 2-35Connector of Gas Dryer.................................................................................................... 2-36Membrane Damage at Gas Dryer ..................................................................................... 2-37

2.9 Open Split ............................................................................................................................... 2-38Inverted Arrangement....................................................................................................... 2-38Need of Open Split ........................................................................................................... 2-38Principle of Open Split ..................................................................................................... 2-39

2.10 Reference Inlet ........................................................................................................................ 2-41Principle of Reference Inlet.............................................................................................. 2-41Dimensions of Capillaries ................................................................................................ 2-42Ref. Purge Valve ............................................................................................................... 2-43

2.11 Reagents .................................................................................................................................. 2-44General Remarks .............................................................................................................. 2-44

Need of Two Separate Reagent Containers ............................................................... 2-44Preparation of Reagent Containers ............................................................................ 2-44Example for Reagent Concentrations ........................................................................ 2-45Waste Disposal of Reagents ....................................................................................... 2-45

Oxidation Reagent ............................................................................................................ 2-46Acid/Catalyst Mixture ...................................................................................................... 2-46

Avoidance of Precipitations ....................................................................................... 2-46Working Without a Catalyst ....................................................................................... 2-46

Avoidance of Organic Solvents ........................................................................................ 2-47Degassing of Reagents ..................................................................................................... 2-47

Need of Reagent Degassing ....................................................................................... 2-47Degassing Procedure .................................................................................................. 2-47

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2.12 Interface Pumps ...................................................................................................................... 2-49General Remarks.............................................................................................................. 2-49Arrangement of Interface Pumps ..................................................................................... 2-49Tubings for Reagent-Pump Connections ......................................................................... 2-50Connecting Eluent Tubings to Pump Heads .................................................................... 2-51Filling the Lines between Containers and Interface Pumps for the First Time ............... 2-52

Suction using a Syringe ............................................................................................. 2-52Suction at the Bottom of Interface Pump .................................................................. 2-52

Motor Blocking at Interface Pumps ................................................................................. 2-52Piston Flush of Interface Pumps ...................................................................................... 2-53

Need of Piston Flush of Interface Pumps .................................................................. 2-53Using Piston Flush Pump for Backpiston Flushing ................................................... 2-53

Adjusting Interface Pumps with HPLC Pump ................................................................. 2-54Removal of Air Bubbles at Pump Heads ................................................................... 2-54

Starting Up Interface Pumps ............................................................................................ 2-56Dismantling Check Valves of Interface Pumps................................................................ 2-57

Step 1 - Replacing and Cleaning Check Valves ........................................................ 2-57Step 2 - Demounting Check Valves ........................................................................... 2-60

Improper Function of Interface Pumps ............................................................................ 2-61Improper Function of Acid Pump .............................................................................. 2-61Improper Function of Oxidation Reagent Pump ....................................................... 2-61

Pulse Damper of Interface Pumps.................................................................................... 2-62

Electronics Area ........................................................................................................................ 3-1

3.1 Individual Electronics Parts...................................................................................................... 3-2USB-RS 232 Converter...................................................................................................... 3-2Manifold Mountable Solenoid Valve ................................................................................. 3-3Power Inlet Module............................................................................................................ 3-4Line Filter........................................................................................................................... 3-5LC IsoLink Control Board ................................................................................................. 3-5Interface Pump Control Boards.......................................................................................... 3-6Switching Power Supply.................................................................................................... 3-8Heater Relay....................................................................................................................... 3-9Voltage Regulator for Cooler ............................................................................................. 3-9

3.2 Comments about Old Electronics ........................................................................................... 3-10Adapter for Old Electronics ............................................................................................. 3-10Operating Two Interfaces................................................................................................. 3-12If Isodat 2.0 Hangs Up with Old Electronics ................................................................... 3-12

3.3 Spare Parts of Electronics....................................................................................................... 3-13

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Isodat 2.0 Software .................................................................................................................... 4-1

4.1 Starting Isodat 2.0 the Very First Time ..................................................................................... 4-2

4.2 Creating a Gas Configuration ................................................................................................... 4-4

4.3 Creating a Configuration........................................................................................................... 4-7Configurations for Autosampler and/or HPLC Pump...................................................... 4-10Editing the Peripheral via Advanced Mode...................................................................... 4-11

Device tab .................................................................................................................. 4-12Events tab ................................................................................................................... 4-14Sequence tab .............................................................................................................. 4-15Monitor Parameter tab ............................................................................................... 4-16System Start Values tab .............................................................................................. 4-17

4.4 LC IsoLink Interface Window ................................................................................................ 4-19Access to LC IsoLink Interface Window ......................................................................... 4-19

If LC IsoLink Interface Window is not Visible ......................................................... 4-20If only Title Bar of LC IsoLink Interface Window is Visible .................................... 4-21

Operations Using LC IsoLink Interface Window............................................................. 4-21Adjusting Standby Parameters ................................................................................... 4-22Changing Flow Rates of Interface Pumps ................................................................. 4-24Starting or Stopping Interface Pumps ........................................................................ 4-25If an Interface Pump is Blocked ................................................................................. 4-25Three-Way Valve (Purge Valve) ................................................................................ 4-26Six-Port Valve ............................................................................................................ 4-27Carrier Gas Flow ........................................................................................................ 4-28Piston Flush ................................................................................................................ 4-30

4.5 Peripheral Visualizations......................................................................................................... 4-32Loading a Peripheral Visualization .................................................................................. 4-32Elements of a Peripheral Visualization............................................................................. 4-33

Storing Graphical Elements ....................................................................................... 4-33Editing Graphical Elements ....................................................................................... 4-34

4.6 Zero Enrichment (Standard On/Off Test)................................................................................ 4-35Instrument......................................................................................................................... 4-35Time Events...................................................................................................................... 4-36Evaluation......................................................................................................................... 4-37Peak Detection.................................................................................................................. 4-37Printout ............................................................................................................................. 4-38ISL Script.......................................................................................................................... 4-38Start Zero Enrichment ...................................................................................................... 4-38Results .............................................................................................................................. 4-39

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4.7 Linearity Test .......................................................................................................................... 4-41

4.8 LC IsoLink Methods............................................................................................................... 4-43Predefined Methods Only as Examples ........................................................................... 4-43Creating a New Method ................................................................................................... 4-44

Instrument tab ............................................................................................................ 4-45Time Events tab ......................................................................................................... 4-46Component Names tab .............................................................................................. 4-48Evaluation tab ............................................................................................................ 4-48Peak Detection tab ..................................................................................................... 4-49Printout tab ................................................................................................................ 4-52ISL Script tab ............................................................................................................. 4-52Saving a Method ........................................................................................................ 4-54

4.9 LC IsoLink Sequences............................................................................................................ 4-56Predefined Sequences Only as Examples ........................................................................ 4-56Creating a New Sequence ................................................................................................ 4-57Saving a New Sequence................................................................................................... 4-59

Save Command .......................................................................................................... 4-59Save As Command .................................................................................................... 4-59Save All Command .................................................................................................... 4-60

Starting a Sequence.......................................................................................................... 4-60Backup in Isodat 2.0......................................................................................................... 4-62Shutdown of Isodat 2.0 .................................................................................................... 4-62

Maintenance .............................................................................................................................. 5-1

5.1 First Startup .............................................................................................................................. 5-2Initial Helium Flushing ...................................................................................................... 5-2Adjusting the Flow Rates................................................................................................... 5-2Leak Check by Initial Argon Measurement ....................................................................... 5-2

Performing Leak Checks ............................................................................................. 5-2Leaks of Gases ............................................................................................................. 5-3Leaks of Liquids .......................................................................................................... 5-3Changing the Flow Rate of HPLC Unit ...................................................................... 5-4Changing the Flow Rates of Reagents ......................................................................... 5-4Waiting for Equilibrium ............................................................................................... 5-4Degassing the Reagents Again .................................................................................... 5-4

Stabilization of CO2 Background ...................................................................................... 5-5

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5.2 Daily Startup ............................................................................................................................. 5-6CO2 Background Scan during Equilibrium (First Time Scan)........................................... 5-6Monitoring CO2 Background by a Second Time Scan (“Five Minutes Test”) .................. 5-7

Principle of “Five Minutes Test“ ................................................................................. 5-7Calculation of Standard Deviation and Slope .............................................................. 5-8

Checking the Other Backgrounds....................................................................................... 5-8Checking Water Background ....................................................................................... 5-8Checking Argon Background ...................................................................................... 5-9Checking Oxygen Background .................................................................................... 5-9

Magnetic Field Scan ........................................................................................................... 5-9Properties of a Magnetic Field Scan ............................................................................ 5-9Adjusting the Parameters for a Magnetic Field Scan ................................................. 5-10

Monitoring Three Different CO2 Background Stati ......................................................... 5-11

5.3 Before Operational Pauses ...................................................................................................... 5-13Before Leaving the System Overnight ............................................................................. 5-13Before Longer Pauses....................................................................................................... 5-13

Switching Off the System .......................................................................................... 5-13Quantitative Removal of Oxidation Reagent ............................................................. 5-13Handling the Interface Pumps .................................................................................... 5-13

Requirements ............................................................................................................................. 6-1

6.1 Site, Electrical and Temperature Requirements ........................................................................ 6-2

6.2 Substance Requirements ........................................................................................................... 6-3Reagents ............................................................................................................................. 6-3Gases................................................................................................................................... 6-3

Compressed Air ........................................................................................................... 6-3

6.3 Hardware and Software Requirements ..................................................................................... 6-5Hardware ............................................................................................................................ 6-5

Available Packages Including Finnigan LC IsoLink ................................................... 6-5IRMS ............................................................................................................................ 6-5

Software.............................................................................................................................. 6-5Minimum Requirements .............................................................................................. 6-5Driver Software for USB Port ...................................................................................... 6-6PCI-GPIB Card and its Driver Software ...................................................................... 6-6Surveyor® HPLC Pump ............................................................................................... 6-6Surveyor® Autosampler ............................................................................................... 6-6

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6.4 Spare Parts Lists ....................................................................................................................... 6-7LC IsoLink Itself (Basic Unit, Part No. 115 5550) ............................................................ 6-7Starter Kit LC IsoLink (Part No 115 6130)........................................................................ 6-7

Installation Kit (Part No. 116 3180) ............................................................................ 6-8

Before Sample Measurement in Direct Injection Mode and HPLC Mode .......................... 7-1

7.1 Introduction .............................................................................................................................. 7-2

7.2 Background Stabilization ......................................................................................................... 7-3General Remarks................................................................................................................ 7-3

Example ....................................................................................................................... 7-3Background Masses as Diagnostic Measures .................................................................... 7-4

CO2 Background (m/z 44) ........................................................................................... 7-4Water Background (m/z 18) ........................................................................................ 7-5Argon Background (m/z 40) ........................................................................................ 7-6Oxygen Background (m/z 32) ..................................................................................... 7-6

7.3 Measuring a Standard Component ........................................................................................... 7-8Reference Gas Peaks.......................................................................................................... 7-8

Adjusting Parameters of Reference Gas Peaks ........................................................... 7-8Starting the Measurement .................................................................................................. 7-9

Recording Reference Gas Peaks .................................................................................. 7-9First Injection of Standard Component Using Direct Injection Mode ...................... 7-10Further Injections of Standard Component Using Direct Injection Mode ................ 7-10Calculation of Standard Deviation ............................................................................ 7-10

Sample Measurement in Direct Injection Mode and HPLC Mode ...................................... 8-1

8.1 Principle.................................................................................................................................... 8-2

8.2 Order of Reference Gas Peaks and Sample Peaks.................................................................... 8-3Injection of Standard Component after Sample Peaks in HPLC Mode............................. 8-5

8.3 Example: Separation of an Amino Acids Mixture Using HPLC Mode ................................... 8-6Principle ............................................................................................................................. 8-6Sample Measurement Results ............................................................................................ 8-6

ThermoELECTRON CORPORATIONviii ______________________Finnigan LC IsoLink Operating Manual_____________________

Read This First

Welcome to the Thermo Electron, Finnigan LC IsoLink Operating Manual!

This Finnigan LC IsoLink Operating Manual describes how to setup and use the Finnigan LC IsoLink.

This Operating Manual includes the following chapters:

Chapter 1: Layout outlines the arrangement of crucial parts by explaining different views onto the interface. Particular emphasis is set on three cabinets, which functionally separate liquid flows area, gas isotope area and electronics area from each other.

Chapter 2: Steps of Analysis describes the important hardware components a sample passes chronologically during its analysis throughout the system.

Chapter 3: Electronics Area deals with electronics components and their arrangement within the cabinet of the interface. It also contains comments about the old electronics and electronics spare parts.

Chapter 4: Isodat 2.0 Software treats of preparing Isodat 2.0 software to control measurements: it denotes how to start Isodat 2.0, how to create a gas configuration, a configuration, a method and a sequence, respectively. These are fundamental requirements to be met before any measurement procedure.

As central features and very helpful control tools, the LC IsoLink Interface window and its underlying peripheral visualization are described in detail.

Furthermore, this chapter denotes fundamental measurements like zero enrichment and linearity test.

Chapter 5: Maintenance begins with routine procedures to be performed at first startup of the interface. Actions to be done at daily startup follow.

Further, this chapter provides information about handling the interface at operational pauses.

Chapter 6: Requirements outlines site, electrical, temperature and substance-related requirements. Furthermore, hard- and software requirements are listed. It ends with various general spare parts lists.

Chapter 7: Before Sample Measurement in Direct Injection Mode and HPLC Mode treats the two procedures to be performed before sample measurement in Direct Injection mode and HPLC mode: background stabilization and standard component measurement.

Chapter 8: Sample Measurement in Direct Injection Mode and HPLC Mode deals with important aspects of sample measurement itself in Direct Injection mode and HPLC mode.

____________________ Finnigan LC IsoLink Operating Manual ______________________ ixThermoELECTRON CORPORATION

Read This FirstChanges to the Manual and Online Help ___________________________________Finnigan LC IsoLink

Changes to the Manual and Online HelpTo suggest changes to this manual or the online Help, please send your comments to:

Thermo Electron (Bremen) GmbHFinnigan Advanced Mass SpectrometryBarkhausenstr. 2D-28197 Bremen

Germany

You are encouraged to report errors or omissions in the text or index. Thank you.

ThermoELECTRON CORPORATIONx________________________Finnigan LC IsoLink Operating Manual_____________________

Read This FirstFinnigan LC IsoLink___________________________________________________________ Abbreviations

AbbreviationsThe following abbreviations are used in this and other manuals and in the online Help.

A ampereac alternating currentADC analog-to-digital converterAP acquisition processorAPCI atmospheric pressure chemical ionizationAPI atmospheric pressure ionizationASCII American Standard Code for Information

Interchangeb bitB byte (8 b)baud rate data transmission speed in events per second°C degrees CelsiusCD compact discCD-ROM compact disc read-only memorycfm cubic feet per minuteCI chemical ionizationCIP carriage and insurance paid tocm centimetercm3 cubic centimeterCPU central processing unit (of a computer)CRC cyclic redundancy checkCRM consecutive reaction monitoring control key on the terminal keyboardd depthDa daltonDAC digital-to-analog converterdc direct currentDDS direct digital synthesizerDEP™ direct exposure probeDS data systemDSP digital signal processor

____________________ Finnigan LC IsoLink Operating Manual ______________________ xiThermoELECTRON CORPORATION

Read This FirstAbbreviations_______________________________________________________Finnigan LC IsoLink

EI electron ionizationEMBL European Molecular Biology Laboratory enter key on the terminal keyboardESD electrostatic dischargeESI electrospray ionizationeV electron voltf femto (10-15)°F degrees Fahrenheit.fasta file extension of a SEQUEST search database fileFOB free on boardft footFTP file transfer protocolg gramG giga (109)GC gas chromatograph; gas chromatographyGC/MS gas chromatograph/mass spectrometerGND electrical groundGPIB general-purpose interface busGUI graphical user interfaceh hourh heightHPLC high-performance liquid chromatographHV high voltageHz hertz (cycles per second)ICIS™ Interactive Chemical Information SystemICL™ Instrument Control Language™ID inside diameterIEC International Electrotechnical CommissionIEEE Institute of Electrical and Electronics Engineersin. inchI/O input/outputk kilo (103, 1000)K kilo (210, 1024)KEGG Kyoto Encyclopedia of Genes and Genomeskg kilogram

ThermoELECTRON CORPORATIONxii_______________________Finnigan LC IsoLink Operating Manual_____________________

Read This FirstFinnigan LC IsoLink___________________________________________________________ Abbreviations

l lengthl literLAN local area networklb poundLC liquid chromatograph; liquid chromatographyLC IRMS liquid chromatography isotope ratio mass

spectrometer

LC/MS liquid chromatograph/mass spectrometerLED light-emitting diodeµ micro (10-6)m meterm milli (10-3)M mega (106)M+ molecular ionMB Megabyte (1048576 bytes)MH+ protonated molecular ionmin minuteml millilitermm millimeterMS mass spectrometer; mass spectrometryMS MSn power: where n = 1MS/MS MSn power: where n = 2MSn MSn power: where n = 1 through 10m/z mass-to-charge ration nano (10-9)NCBI National Center for Biotechnology Information

(USA)NIST National Institute of Standards and Technology

(USA)OD outside diameterΩ ohmp pico (10-12)Pa pascalPCB printed circuit boardPID proportional / integral / differentialP/N part number

____________________ Finnigan LC IsoLink Operating Manual _____________________ xiiiThermoELECTRON CORPORATION

Read This FirstAbbreviations_______________________________________________________Finnigan LC IsoLink

P/P peak-to-peak voltageppm parts per millionpsig pounds per square inch, gaugeRAM random access memoryRF radio frequencyRMS root mean squareROM read-only memoryRS-232 industry standard for serial communicationss secondSIM selected ion monitoringsolids probe direct insertion probeSRM selected reaction monitoringss stainless steelSSQ® single stage quadrupoleTCP/IP transmission control protocol / Internet protocolTIC total ion currentTorr torrTSQ® triple stage quadrupoleu atomic mass unitURL uniform resource locatorV voltV ac volts alternating currentV dc volts direct currentvol volumew widthW wattWWW World Wide Web

Note. Exponents are written as superscripts. In the corresponding online Help, exponents are sometimes written with a caret (^) or with e notation because of design constraints in the online Help. For example:

MSn (in this manual) Ms^n (in the online Help)

105 (in this manual) 10^5 (in the online Help)

ThermoELECTRON CORPORATIONxiv ______________________Finnigan LC IsoLink Operating Manual_____________________

Read This FirstFinnigan LC IsoLink_________________________________________________Typographical Conventions

Typographical ConventionsTypographical conventions have been established for Thermo Electron San Jose manuals for the following:

• Data input

• Boxed information

• Topic headings

Data InputThroughout this manual, the following conventions indicate data input and output via the computer:

• Messages displayed on the screen are represented by capitalizing the initial letter of each word and by italicizing each word.

• Input that you enter by keyboard is represented in bold face letters. (Titles of topics, chapters, and manuals also appear in bold face letters.)

• For brevity, expressions such as “choose File > Directories” are used rather than “pull down the File menu and choose Directories.”

• Any command enclosed in angle brackets < > represents a single keystroke. For example, “press ” means press the key labeled F1.

• Any command that requires pressing two or more keys simultaneously is shown with a plus sign connecting the keys. For example, “press + ” means press and hold the key and then press the key.

• Any button that you click on the screen is represented in bold face letters and a different font. For example, “click on Close”.

____________________ Finnigan LC IsoLink Operating Manual _____________________ xvThermoELECTRON CORPORATION

Read This FirstTypographical Conventions_____________________________________________Finnigan LC IsoLink

Boxed InformationInformation that is important, but not part of the main flow of text, is displayed in a box such as the one below.

Boxed information can be of the following types:

• Note – information that can affect the quality of your data. In addition, notes often contain information that you might need if you are having trouble.

• Caution – information necessary to protect your instrument from damage.

• Warning – hazards to human beings. Each Warning is accompanied by a Warning symbol.

Note. Boxes such as this are used to display information.

ThermoELECTRON CORPORATIONxvi ______________________Finnigan LC IsoLink Operating Manual_____________________

Read This FirstFinnigan LC IsoLink_________________________________________________Typographical Conventions

Topic HeadingsThe following headings are used to show the organization of topics within a chapter:

Chapter 1Chapter Name

1.2 Second Level Topics

Third Level Topics

Fourth Level Topics

Fifth Level Topics

____________________ Finnigan LC IsoLink Operating Manual ____________________ xviiThermoELECTRON CORPORATION

Read This FirstReply Cards________________________________________________________Finnigan LC IsoLink

Reply CardsThermo Electron San Jose manuals contain one or two reply cards. All manuals contain a Customer Registration / Reader Survey card and some contain a Change of Location card. These cards are located at the front of each manual.

The Customer Registration / Reader Survey card has two functions. First, when you return the card, you are placed on the Thermo Electron San Jose mailing list. As a member of this list, you receive application reports and technical reports in your area of interest, and you are notified of events of interest, such as user meetings. Second, it allows you to tell us what you like and do not like about the manual.

The Change of Location card allows us to track the whereabouts of the instrument. Fill out and return the card if you move the instrument to another site within your company or if you sell the instrument. Occasionally, we need to notify owners of our products about safety or other issues.

ThermoELECTRON CORPORATIONxviii _____________________Finnigan LC IsoLink Operating Manual_____________________

Chapter 1Layout

1.1 Overall View

1.2 Front View

1.3 Functional Separation into Cabinets

1.4 Back View

1.5 Right View

1.6 Left View

____________________ Finnigan LC IsoLink Operating Manual ____________________ 1-1ThermoELECTRON CORPORATION

LayoutOverall View _______________________________________________________Finnigan LC IsoLink

1.1 Overall View

Figure 1-1. Finnigan LC IsoLink

ThermoELECTRON CORPORATION1-2 _____________________Finnigan LC IsoLink Operating Manual_____________________

LayoutFinnigan LC IsoLink____________________________________________________________ Overall View

Figure 1-2. Finnigan LC IsoLink and Finnigan Surveyor® HPLC Unit


LayoutOverall View _______________________________________________________Finnigan LC IsoLink

Figure 1-3. Finnigan LC IsoLink, Finnigan Surveyor® HPLC Unit and Finnigan Deltaplus Advantage


LayoutFinnigan LC IsoLink______________________________________________________________ Front View

1.2 Front View

1. Front doorOpen it by grasping into the cavity at its bottom side (see arrow) and then pulling it outwards as shown in Figure 1-5. This gives comfortable access to the pump heads located behind it. The pump heads are easily accessible,

• when connecting to oxidation reagent and acid, respectively and

• when replacing the pump heads(see e.g. Removal of Air Bubbles at Pump Heads on page 2-54).

Figure 1-4. Front View of Finnigan LC IsoLink

1

2

4

3

6

5

78

9

10

11

14

12

13

15


LayoutFront View _________________________________________________________Finnigan LC IsoLink

Two cut-outs in the door, one at its upper edge and the other one at its lower edge, allow to stick through the inlet tubes to the pumps. In doing so, the door can be kept closed during operation.

2. Oven temperature control (by JUMO itron 16 temperature controller)

3. Gas regulator for carrier gas (He)

4. Gas regulator for purge gas (He)

5. Gas regulator for reference gas (CO2)

6. Inject button. This pushbutton switch turns the two-position six-port valve from Load position to Inject position and vice versa.Load position: The yellow lamp 8 is flashing. A solvent or sample can be injected into the sample loop by a HPLC syringe. Use the external needle port, which - as the sample loop - needs to be screwed on.

Inject position: The green lamp 7 is flashing. The content within the sample loop will reach the interface via a PEEK™ line(see e.g. Backflush of Separation Unit Capillaries on page 2-29).

7. The green lamp flashes during Inject position.

8. The yellow lamp flashes during Load position.

9. Two-position six-port valve(electronically switching, see Figure 2-7 and Figure 2-8).

10. Manometer for reference gas (CO2)

11. Manometer for purge gas (He)

12. Manometer for carrier gas (He)

13. Ref Purge (purge reference gas valve)

14. Connection between six-port valve and interface(PEEK™ tube, o.d. = 1/16”, i.d. = 0.01 mm)

15. Protective lid

Figure 1-5. Opening the Front Door of Finnigan LC IsoLink


LayoutFinnigan LC IsoLink______________________________________________________________ Front View

Warning. Pay attention to the safety instructions (Figure 1-6, Figure 1-7):

• Always operate Finnigan LC IsoLink with its protective lid attached so that the system is closed! Otherwise, you run the risk of an electric shock as the device is operated at 230 V mains voltage!

• If you temporarily want to remove its lid, always pull out the power plug and turn off the main switch before!

• To remove the lid then, first unscrew the screws at the rear panel. Then lift up the lid a bit until it slips out of its mounting (at the cone). Finally, move it slightly backwards.

• Only trained personnel may remove the lid and open the device!

Figure 1-6. Safety Instruction at Rear Panel

Figure 1-7. Rear Panel


LayoutFront View _________________________________________________________Finnigan LC IsoLink

Figure 1-8. Front View of Finnigan LC IsoLink (Door Removed)

1 six-port valve

2 feedthrough to interface

3 yellow LED (Load position)

4 green LED (Inject position)

5 Injection button

6 Ref Gas manometer/regulator

7 He purge manometer/regulator

8 He carrier manometer/regulator

9 Ref Gas purge

10 JUMO itron 16 temperature controller

11 acid pump

12 oxidation reagent pump

13 feedthrough piston flush

1

3

2

4

6b

5

6a

7a 7b

8a 8b

109

12

11

13


LayoutFinnigan LC IsoLink__________________________________________ Functional Separation into Cabinets

1.3 Functional Separation into Cabinets

As shown in Figure 1-9 and Figure 1-10, a mid plate and a separating plate form three different cabinets within Finnigan LC IsoLink. Each cabinet houses its own functional unit:

• electronics area

• liquid flows area

• gas isotope area

Figure 1-9. Plates and Cabinets within Finnigan LC IsoLink

pump heads

front panel

mid plate

separating plate

rear panel

liquid flows area

electronics area

gas isotope area


LayoutFunctional Separation into Cabinets ______________________________________Finnigan LC IsoLink

Figure 1-10. Functional Separation into Three Cabinets (Top View)

Back

Front

electronics area

liquid flows area

oxidant pumpacid pump

pulse damper

backp. flush pump

gas isotope area

oxidation reactor

cooler

separation unit

open split

reference inlet

2 water traps

separating plate

mid plate

1. The liquid flows area houses:

• interface pumps (that is, oxidation reagent pump and acid pump)

• pulse damper

• piston flush pump

2. In the electronics area, that is the cabinet behind the first one, electronics is mounted.

3. The gas isotope area is separated from both other cabinets by the mid plate. It houses:

• oxidation reactor

• cooler

• separation unit

• open split and reference inlet

• two water traps

ThermoELECTRON CORPORATION1-10 ____________________Finnigan LC IsoLink Operating Manual_____________________

LayoutFinnigan LC IsoLink______________________________________________________________ Back View

1.4 Back View

In order to enhance mechanical stability, the rear panel is a bit narrower than the front panel. The rear panel contains vent holes (see 1 in Figure 1-11) and serves as a feedthrough for the

• electronics (i.e. USB connector, power cable, connection to the IRMS; see 5, 9, 14 in Figure 1-11)

Figure 1-11. Rear Panel of Finnigan LC IsoLink (Housing)

1 vent holes

2 waste line of purge of interface pumps(via a Swagelok® connector SS-200-1 to a SS capillary)

3 exit of piston flush(Teflon® tubing)

4 waste line of interface’s separation unit(via a Swagelok® connector SS-200-1)

5 USB connector

6 exit of IRMS capillary

7 fan

power supply unit (8 - 10)

8 two fuses

9 connector for power plug

10 main switch (on/off)

gas inlet pipes (11 - 13)

11 compressed air inlet

12 Ref Gas inlet (CO2)

13 He inlet

14 cable feedthrough to IRMS

15 grounding (ground bolt)

11

9

23

4

1213

5

1

15

8 10

14

7

6

____________________ Finnigan LC IsoLink Operating Manual ___________________ 1-11ThermoELECTRON CORPORATION

LayoutBack View _________________________________________________________Finnigan LC IsoLink

• gases on their way into the interface (that is, reference gas and helium; see 11, 12, 13 in Figure 1-11)

• waste line (coming from the membrane of the separation unit and from purging oxidation reagent pump and acid pump; see 2, 4 in Figure 1-11).

Figure 1-12. Back View onto Gas Isotope Area


LayoutFinnigan LC IsoLink______________________________________________________________ Right View

1.5 Right View• The parts related to the liquids flow are arranged in the left cabinet

(liquid flows area).

• The electronic parts are housed in the right cabinet (electronics area).

• The three-way valve, that is “purge valve“ 2 is driven by a pneumatic actuating drive (compressed air). It is used to purge the interface pumps, that is acid pump 3 and oxidation reagent pump 4.See Three-Way Valve (Purge Valve) on page 4-26.

Figure 1-13. Right View onto Liquid Flows Area and Electronics Area

liquid flows area

electronics area

1a

1b

2

3

4

56

10

11

14

8; 9

12

7

16

13

1 cut-outs (1a, 1b)

2 three-way valve(“purge valve”)

3 acid pump

4 ox. reagent pump

5 pulse damper

6 piston flush pump

7 terminal board (24 V)

8 control board(ox. reagent pump)

9 control board(acid pump)

10 USB-RS 232 converter

11 LC IsoLink control board

12 mains switch (on/off)

13 valve cluster

14 fan

15 relay filter(for temp. controller)

16 terminal board

17 line filter (230 V)

18 purge outlet

19 power supply (24 V)

1516

17

1819


LayoutRight View _________________________________________________________Finnigan LC IsoLink

• The control board for the oxidation reagent pump 8 and the control board for the acid pump 9 are arranged back-to-back (Figure 3-11).

• The USB-RS 232 converter controls the interface pumps via computer. See USB-RS 232 Converter on page 3-2.

• The LC IsoLink control board 11 controls piston flush pump 6, valve cluster 13 and six-port valve. See Figure 3-5 and Figure 3-8.

• A fan 14 cools the interface and provides stable temperature.


LayoutFinnigan LC IsoLink_______________________________________________________________ Left View

1.6 Left View

Figure 1-14. Left View onto Gas Isotope Area

1 connector separation unit - waste line

2 t-piece

3 oxidation reactor

4 cooler (fan, on a mounting plate)

5 separation unit(membrane exchanger)

6 gas dryer(two water traps)

7 open split(for the interface)

8 reference inlet(for Ref Gas)

9 waste

10 six-port valve

11 Ref Gas

12 He (purge)

13 He (carrier)

14 JUMO itron 16 temperature controller

15 reducer

1

2

3

4

5

6b6a

78

15

9

10

12

11

14

13


Chapter 2Steps of Analysis

2.1 Range of Applications and Analysis Principle

2.2 HPLC Unit

2.3 Six-Port Valve - Leaving the HPLC Unit and Entering the Interface

2.4 Sample Loop and HPLC Syringe

2.5 T-Piece

2.6 Oxidation Reactor

2.7 Separation Unit

2.8 Gas Dryer

2.9 Open Split

2.10 Reference Inlet

2.11 Reagents

2.12 Interface Pumps


Steps of AnalysisRange of Applications and Analysis Principle ________________________________Finnigan LC IsoLink

2.1 Range of Applications and Analysis Principle

Range of ApplicationsFinnigan LC IsoLink is built to directly address analysis of polar and thermo-labile compounds. Most of these compounds (such as amino acids, peptides, carbohydrates, nucleotides, alcohols, organic acids, sulfa drugs and many more) can be chromatographically separated in aqueous phase using inorganic buffers on ion exchange, size exclusion or specific reversed phase HPLC columns. Finnigan LC IsoLink enables on-line high precision determination of the 13C/12C isotope ratio of each separated component. It offers a wide range of applications:

1. Metabolism studies

• Tracer experiments with amino acids, lipids and carbohydrates

• Nucleotide analysis for DNA synthesis rates

2. Food and food ingredients

• Adulteration control of honey and fruit juices

• Determinating the origin of flavors and nutrients

• Analysis of phospholipids, carbohydrates, amino acids and vitamins

3. Pharmaceuticals

• Authenticity control of drugs

• Investigation of agents and additives

4. Doping control

• Analysis of steroids and hormones

5. Biogeochemistry

• History of biomolecules in sediments and soils

Analysis PrincipleFinnigan LC IsoLink is characterized by oxidation of individual organic compounds still in an aqueous phase. Because the oxidation happens in the mobile phase, it is restricted to water that may additionally contain inorganic buffers. Otherwise, the mobile phase would be converted to CO2 as well.

Afterwards, the mobile phase is separated from the formed CO2 by the separation unit. This process is quantitative and fractionation-free.


Steps of AnalysisFinnigan LC IsoLink___________________________________ Range of Applications and Analysis Principle

Finnigan LC IsoLink can be used in either of two injection modes, one for compound-specific isotope analysis and one for bulk-stable isotope analysis:

• In HPLC mode, samples are injected by a loop injection valve in front of the HPLC column. The mixture of organic compounds in the sample is separated on the HPLC column and the constant flow of the mobile phase is maintained through the oxidation interface.

• The Direct Injection mode (µ-EA mode) is a fast analysis of all water soluble materials and is thus a bulk measurement. Reference materials and unknown bulk samples can be analyzed with unmatched sensitivity and speed. Samples can be injected via the needle port into a sample loop of variable size, which is placed at the six-port-valve behind the HPLC column.

At the t-piece, a constant mixture of reagents is added to the mobile phase. The oxidation reagent consists of a peroxodisulfate M2S2O8 (M+ = Na+, K+, NH4+) in water, the acid/catalyst reagent of a mixture of phosphoric acid H3PO4 and silver nitrate AgNO3. Both reagents are pumped separately by two-head pumps.

To avoid pulsations of the reagent streams, which would result in an unstable background signal, a pulse damper is placed behind the two-head pumps.

The mixture of reagents and mobile phase flows through a capillary oxidation reactor at 99 ºC. Here, all organic compounds are converted individually and quantitatively into CO2 without isotope fractionation:

The oxidation potential of the oxidant can be increased by the addition of acid. Furthermore, the oxidative reaction can be catalyzed by Ag+ ions.

Figure 2-1. Analysis Principle of Finnigan LC IsoLink

HPLC

IRMS

Gas Dryer OpenSplit

He He+H20

OxidationReactor

Separationof the mobile phase

OxidationReagents

C12H22O11 C6H12O6 CO2 CO2 CO2

Figure 2-2. Oxidation of an Organic Compound to CO2

6 S2O82- C2H5OH 3 H2O+ + 12 SO42- 2 CO2 12 H

++ +→


Steps of AnalysisRange of Applications and Analysis Principle ________________________________Finnigan LC IsoLink

After passing the oxidation reactor, the mobile phase is cooled. The individual CO2 peaks are separated from the liquid phase by a separation unit: the CO2 is transferred through thin membranes into a counter flow of helium. The use of membranes, the pressure difference and the difference in CO2 concentration between inside and outside of the membranes contribute to an almost complete degassing of the liquid phase.

Acidification of the liquid can also be useful for complete degassing as it decreases solubility of CO2 in the liquid phase and thus shifts the chemical equilibrium of Figure 2-4 to the left side.

The solubility of CO2 is given by the Henry coefficient H1,2 (1 gas, 2 solvent),

with fugacity f and mole fraction x of the dissolved CO2. For the solubility of CO2 in water, the Henry coefficient at 298.15 K is 1622 bar. H1,2 increases with small amounts of acid. Subsequently, the solubility of CO2 in water decreases. Furthermore, solved salts, e.g. (NH4)2SO4, also increase H1,2 and assist degassing in addition by salt-out effects.

The individual CO2 peaks entrained in the helium stream are subsequently dried in an Nafion® on-line gas drying unit and then admitted to the IRMS via an open split.

These analysis steps are rather comprehensible and will be described below according to Figure 2-5, the schematic diagram of Finnigan LC IsoLink. Any of the parts described below is part of the interface. Only the HPLC unit is arranged outside of it. Besides, the oxidation reagent container and the acid/catalyst container are usually placed on top of it (see 4 in Figure 2-6).

Figure 2-3. Definition of Henry Coefficient H1,2

Figure 2-4. Chemical Equilibrium of CO2 in Water

H1,2f1x1-----

x1 0→

x2 1→

T const.=

lim=

CO2 H2O+ HCO-3 H

++←


Steps of AnalysisFinnigan LC IsoLink __________________________________ Range of Applications and Analysis Principle

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____________________ Finnigan LC IsoLink Operating Manual_____________________ 2-5ThermoELECTRON CORPORATION

Steps of AnalysisHPLC Unit _________________________________________________________Finnigan LC IsoLink

2.2 HPLC Unit

Purging HPLC Pump and HPLC Pulse DamperThe HPLC pulse damper is used to minimize the pulsations of the HPLC pump. Thereby, it ensures a smooth background signal. A membrane (“diaphragm“, not semipermeable) is located at its backside. Behind the membrane, a distorted steel capillary filled with water is located.

Figure 2-6. Parts of Finnigan Surveyor® HPLC Unit

Note. Refer to Surveyor® MS Pump Hardware Manual, Revision A, Part No. 60053-97042 (particularly to its chapter 5 - Maintenance) for:

• operating, dismantling and subsequently cleaning the HPLC pump

• replacing the Liquid Displacement Assembly, LDA

The HPLC pump produces pulsations as well. After a long time of operation, the LDA must be replaced in order to restrict the wiggles of the HPLC pump.

HPLCUnit

3

4

1

2

The HPLC unit is arranged outside of Finnigan LC IsoLink interface.

As shown in Figure 2-6, it comprises:

• HPLC pump 1

• Autosampler 2

• UV detector 3

• Reagent containers 4

The oxidation reagent container and the acid/catalyst mixture container can be placed on top of the HPLC unit.


Steps of AnalysisFinnigan LC IsoLink______________________________________________________________ HPLC Unit

For trouble-free operation, purge HPLC pump and HPLC pulse damper from time to time. This helps to remove possible air bubbles and organic contaminations, which otherwise are easily deposited especially at the diaphragm and thus increase the CO2 background.

For details, refer to Surveyor® MS Pump Hardware Manual, Revision A, Part No. 60053-97042.

Usage of Pure WaterFor proper handling of the HPLC unit, the following problems concerning the water in use must be solved:

• Any organic substances must be eliminated from the water.

• Water must be sufficiently degassed.

• Regassing of water must be avoided.

• Particles must be kept away from the system by eluent filters.

Elimination of Organic Substances

General Remarks

As organic substances, e.g. bacteria and residuals of solvents can occur. They must all be quantitatively eliminated from the water.

Note. For details about the HPLC pulse damper refer to Surveyor® MS Pump Hardware Manual, Part No. 60053-97042 (particularly to its chapter 5 - Maintenance).

Warning. First, open the purge valve in order not to damage the system before applying a high flow!

Note. Never contaminate any parts of the HPLC unit with organic materials, e.g. the Surveyor® MS pump!

Warning. In order not to damage the HPLC column remove it before eliminating any organic substances!



Elimination of BacteriaBacteria tend to spread in aqueous solutions and settle particularly on semipermeable membranes. They will be subject to oxidation as well and thereby markedly increase the CO2 background signal.

If the CO2 background signal nevertheless is still considerably high after purging with water, eliminate remaining bacteria by oxidation: therefore, purge the HPLC unit including the HPLC pulse damper with oxidation reagent daily before measurement (see Purging HPLC Pump and HPLC Pulse Damper on page 2-6).

A lower concentration of M2S2O8 (M+ = Na+, K+, NH4+) than that used for measurements is sufficient, e.g. 50-100 g/l. Finally, purge several times using pure water.

Bacteria will have increased their populations especially after longer working pauses or weekends. Therefore, when restarting operation after such pauses purge even longer.

Warning. For trouble-free operation, purge the system daily including the HPLC pulse damper. Purging helps to remove possible air bubbles and organic contaminations.

• First, open the purge valve in order not to damage the system before applying a high flow!

• Always begin purging by using only water.

For details, refer to Surveyor® MS Pump Hardware Manual, Revision A, Part No. 60053-97042.

Warning. Let the oxidation reagent pass neither through the HPLC column nor through the interface! Instead, use the purge valve of the HPLC pulse damper to let pass oxidation reagent as follows:

• Suck in the oxidation reagent.

• Open the purge valve at the HPLC pulse damper.

• Purge using oxidation reagent.

This procedure allows to apply relatively high flow rates.


Steps of AnalysisFinnigan LC IsoLink______________________________________________________________ HPLC Unit

Avoidance of Organic Solvents

Release of Organic Substances from the HPLC ColumnAs usual when working with organic- or polymer-containing HPLC columns, small amounts of organic substances can be released from the column and transported into the system (bleeding of HPLC column). They will increase the background signal and can block the flow through the separation unit.

Water Degassing

Warning. Always run the HPLC pump only with water as the mobile phase!

• Ensure that the HPLC pump has never been operated using any organic solvent before!

• Organic solvents are easily oxidized. Furthermore, they can contaminate the seals of its pistons. Each pump movement could be visible as high background and/or pulse at the corresponding mass.

• If the HPLC pump has been contaminated by any organic solvent, it needs to be dismantled and then intensely be cleaned: only use water, an ultrasonic bath and replace the seals.

Note. Therefore,

• an in-line filter is located between HPLC column and six-port valve. It prevents particles that have been released from the bleeding HPLC column from spreading within the system. See 4 in Figure 6-7.

• only use HPLC columns that release negligible amounts of organic substances (that is, marginal bleeding).

• only use HPLC columns that can be run with water as a solvent (additionally containing inorganic buffers).

• use the HPLC column that is suited best for your particular application.

Note. The water used as the only solvent must be sufficiently degassed!

• First, degas in an ultrasonic bath.

• Afterwards, strip the water using a helium atmosphere to prevent regassing of the water.

• Finally, leave the water under this atmosphere by providing a constant helium stream.

Note. Change the water at least every three days.

Additionally, slightly acidify it to eliminate possible organic material via protonation.



Avoidance of Water Regassing

Keeping Particles away by Eluent FiltersOxidation reagent container and acid/catalyst cont

Documents

Finnigan LC IsoLinksilab/ThermoManuals/LC/115_6140_LC... · 2008. 10. 14. · _____ Finnigan LC IsoLink Operating Manual_____ a Thermo ELECTRON CORPORATION Safety and EMC Information

Finnigan LC IsoLinksilab/ThermoManuals/LC/115_6140_LC... · 2008. 10. 14. · _ Finnigan LC IsoLink Operating Manual_ a Thermo ELECTRON CORPORATION Safety and EMC Information