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PFC Analysis Kit for ACQUITY UPLC

System Guide

71500183002/Revision A

Copyright © Waters Corporation 2009 All rights reserved

Copyright notice

© 2009 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.

Trademarks

ACQUITY, ACQUITY UPLC, Oasis, UltraPerformance LC, UPLC, and Waters are registered trademarks of Waters Corporation, and IntelliStart, MassLynx, PFC Isolator, QuanLynx, Quattro micro, Quattro Premier, TargetLynx, and “THE SCIENCE OF WHAT’S POSSIBLE.” are trademarks of Waters Corporation.PharMed and Tygon are registered trademarks of Saint-Gobain Ceramics & Plastics, Inc.Teflon is a registered trademark of E.I. du Pont de Nemours & Co., Inc.PEEK is a trademark of Victrex plc.Other registered trademarks or trademarks are the sole property of their owners.

Customer comments

Waters’ 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].

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Contacting Waters

Contact Waters® with enhancement requests or technical questions regarding the use, transportation, removal, or disposal of any Waters product. You can reach us via the Internet, telephone, or conventional mail.

Safety considerations

Some reagents and samples used with Waters instruments and devices can pose chemical, biological, and radiological hazards. You must know the potentially hazardous effects of all substances you work with. Always follow

Waters contact information

Contacting medium InformationInternet The Waters Web site includes contact information

for Waters locations worldwide. Visit www.waters.com, and click Waters Division > Contact Waters.

Telephone and fax From the USA or Canada, phone 800 252-HPLC, or fax 508 872 1990.For other locations worldwide, phone and fax numbers appear in the Waters Web site.

Conventional mail Waters Corporation34 Maple StreetMilford, MA 01757USA

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Good Laboratory Practice, 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.

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

Using this kit

When using this kit, follow standard quality-control (QC) procedures and the guidelines presented in this section.

Applicable symbols

Caution: Read the system guide thoroughly before using the product, and follow the instructions carefully. Waters cannot guarantee reliable assay results if you deviate from these instructions.

Caution: Do not use the kit components if the kit’s packaging is opened or damaged on arrival.

Symbol DefinitionConfirms that a manufactured product complies with all applicable European Community directivesAustralia C-Tick EMC Compliant

Confirms that a manufactured product complies with all applicable United States and Canadian safety requirements

ABN 49 065 444 751

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Audience and purposeThis document is for laboratory personnel. It includes procedures and information for installing, operating, and maintaining the PFC Analysis Kit for ACQUITY UPLC® systems.

Intended use of the PFC analysis kitWaters designed the PFC Analysis Kit for ACQUITY UPLC systems to analyze trace amounts of perfluorinated compounds in surface water samples.

CalibratingTo calibrate LC systems, follow acceptable calibration methods using at least five standards to generate a standard curve. The concentration range for standards must include the entire range of QC samples, typical specimens, and atypical specimens.When calibrating mass spectrometers, consult the calibration section of the operator’s guide for the instrument you are calibrating. In cases where an overview and maintenance guide, not operator’s guide, accompanies the instrument, consult the instrument’s online Help system for calibration instructions.

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

ISM classification

ISM Classification: ISM Group 1 Class BThis classification has been assigned in accordance with CISPR 11 Industrial Scientific and Medical (ISM) instruments requirements. Group 1 products apply to intentionally generated and/or used conductively coupled

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radio-frequency energy that is necessary for the internal functioning of the equipment. Class B products are suitable for use in both commercial and residential locations and can be directly connected to a low voltage, power-supply network.

EC Authorized Representative

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

Telephone: +44-161-946-2400Fax: +44-161-946-2480Contact: Quality manager

vi

Table of Contents

Copyright notice ................................................................................................... ii

Trademarks ............................................................................................................ ii

Customer comments ............................................................................................. ii

Contacting Waters ............................................................................................... iii

Safety considerations .......................................................................................... iii Safety advisories ................................................................................................. iv

Using this kit ......................................................................................................... iv Applicable symbols ............................................................................................. iv Audience and purpose.......................................................................................... v Intended use of the PFC analysis kit.................................................................. v Calibrating ........................................................................................................... v Quality-control ..................................................................................................... v

ISM classification .................................................................................................. v ISM Classification: ISM Group 1 Class B .......................................................... v

EC Authorized Representative ......................................................................... vi

1 PFC analysis kit ..................................................................................... 1-1

Kit contents ....................................................................................................... 1-2 ACQUITY UPLC retrofit kit for PFC analysis .............................................. 1-3 PFC standards ................................................................................................. 1-3

Purpose ................................................................................................................ 1-6

Compatible ACQUITY UPLC system configurations ................................ 1-7

Software control ................................................................................................ 1-7

Table of Contents vii

2 Setting up the System Hardware ....................................................... 2-1

Preparing an existing system ........................................................................ 2-3

Preparing the solvent reservoirs ................................................................. 2-5

Installing the columns and tubing ................................................................ 2-7

Removing the PFC analysis components .................................................. 2-10

3 Preparing the System for Analysis .................................................... 3-1

Flushing the system .......................................................................................... 3-2

Preparing eluents and wash solvents .......................................................... 3-3

4 Verifying System Performance ........................................................... 4-1

Preparing the standards ................................................................................. 4-2

Performing the verification test ................................................................... 4-2 Opening the project file ................................................................................... 4-3

Interpreting the test data ............................................................................... 4-9

5 Preparing Samples ................................................................................ 5-1

Typical sample preparation procedure ...................................................... 5-1

6 Troubleshooting ..................................................................................... 6-1

PFC analysis troubleshooting ........................................................................ 6-2

Thorough cleaning of solvent reservoirs .................................................... 6-4

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

Warning symbols ............................................................................................... A-1 Task-specific hazard warnings........................................................................ A-1 Task-specific hazard warnings........................................................................ A-2 Warnings that apply to particular instruments, instrument components, and

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

viii Table of Contents

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

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

Electrical and handling symbols ................................................................. A-12 Electrical symbols .......................................................................................... A-12 Handling symbols .......................................................................................... A-13

Index ..................................................................................................... Index-1

Table of Contents ix

x Table of Contents

1 PFC analysis kit

Use the kit to quantify trace levels of perfluorinated compounds (PFCs) in surface water samples. The kit minimizes contamination from system components and separates background contaminants from the analytes of interest. Contents

Topic PageKit contents 1-2Purpose 1-6Compatible ACQUITY UPLC system configurations 1-7Software control 1-7

1-1

Kit contents

The PFC analysis kit includes the elements described in the table, below. Some elements are described in subsequent sections of this guide.

Caution: To help minimize PFC contamination and bacterial growth in the system’s solvent supply, follow the recommendations in this guide for appropriate glassware and bottle closures.

PFC analysis kit contents

Part Description Analytical column ACQUITY UPLC® BEH C18 2.1 ×

50-mm analytical column ACQUITY UPLC retrofit kit for PFC analysis

See table titled “Retrofit kit for PFC analysis” on page 1-3.

PFC standards See “PFC standards” on page 1-3.

Oasis® HLB PLUS extraction cartridges, 225 mg/60 μm

Box of 50

Vials, 12 × 32-mm, 2-mL, polypropylene, screw-neck, with polyethylene, septumless caps

Pack of 100

Borosilicate glass solvent reservoirs • Clear, 500-mL • Clear, 1000-mL (2) • Amber, 500-mL (2) • Amber, 1000-mL (2)

High-density polyethylene solvent caps, with accessories

• Solvent-cap inner liners (7) • Closed-thread solvent caps

(7 colors) • Open-thread solvent caps

(7 colors) • Solvent-cap-liner plugs (14) • Filters, 10-μm (7)

1-2 PFC analysis kit

ACQUITY UPLC retrofit kit for PFC analysisUsing the retrofit kit, you can install a complete flow path suitable for the trace analysis of perfluorinated compounds.

PFC standardsThe standards are divided into two groups: analytical standards used for calibration, and preparation standards used for quality control.

PFC calibration standards

For analytical standards, the PFC kit supplies five PFC calibration standards, one PFC internal standard, and one PFC recovery standard. For quality control, five preparation standards are provided.

Retrofit kit for PFC analysis

Part Description

PFC Isolator™ column assembly 2.1 × 50-mm

Tubing assembly, mixer to isolator column Coiled stainless-steel Tubing assembly, valve to isolator column Stainless-steel Tubing assembly, column to detector PEEK™

Tubing assemblies, solvent inlet PEEK (set of 7) Nut extender Flangeless, shortSample loop 10-μL

Analytical standards for calibration

Label Concentration VialPFC Calibration Standard #1

5.0 ng/mL for each analyte

2-mL, flame-sealed ampule

PFC Calibration Standard #2

25 ng/mL for each analyte





Kit contents 1-3

PFC Calibration standards #1 through #5 contain the following analytes:







Analytes in calibration standards

Analyte Chemical formula CAS number1

1. CAS (Chemical Abstracts Service, a division of the American Chemical Society) numbers are unique identifiers for chemical substances.

Perfluorobutyric acid CF3(CF2)2CO2H 375-22-4Perfluoropentanoic acid CF3(CF2)3CO2H 2706-90-3Perfluorohexanoic acid CF3(CF2)4CO2H 307-24-4Perfluoroheptanoic acid CF3(CF2)5CO2H 375-85-9Perfluorooctanoic acid CF3(CF2)6CO2H 335-67-1Perfluorononanoic acid CF3(CF2)7CO2H 375-95-1Perfluorodecanoic acid CF3(CF2)8CO2H 335-76-2Perfluoroundecanoic acid CF3(CF2)9CO2H 2058-94-8 Perfluorododecanoic acid CF3(CF2)10CO2H 307-55-1Perfluorotridecanoic acid CF3(CF2)11CO2H 72629-94-8Perfluorotetradecanoic acid CF3(CF2)12CO2H 376-06-7Perfluorobutanesulfonic acid CF3(CF2)3SO3H 375-73-5Perfluorohexanesulfonic acid CF3(CF2)5SO3H 355-46-4Perfluoroheptanesulfonic acid CF3(CF2)6SO3H Not availablePerfluorooctanesulfonic acid CF3(CF2)7SO3H 1763-23-1Perfluorodecanesulfonic acid CF3(CF2)9SO3H 335-77-3

Analytical standards for calibration (Continued)

Label Concentration Vial


Internal and recovery standards for calibration

Label Analytes Concentration VialPFC Internal Standard

• Perfluorooctanoic acid (1,2,3,4-13C4)

• Perfluorooctanesulfonic acid (1,2,3,4-13C4)



PFC Recovery Standard

• Perfluorohexanoic acid (1,2-13C2)

• Perfluoro-n-[1,2-13C2]decanoic acid

• Perfluorohexanesulfonic acid (18O2)



Preparation standards for quality control

Label Analytes Concentration VialsPFC Quality Control Standards

• Perfluoropentanoic acid• Perfluorohexanoic acid• Perfluoroheptanoic acid• Perfluorooctanoic acid• Perfluorononanoic acid• Perfluorodecanoic acid• Perfluoroundecanoic acid• Perfluorododecanoic acid• Perfluorobutanesulfonic acid• Perfluorohexanesulfonic acid• Perfluorooctanesulfonic acid• Perfluoroheptanesulfonic acid• Perfluorodecanesulfonic acid

5.0 to 500 ng/mL for each analyte

Five, 2-mL, flame- sealed ampules

Kit contents 1-5

Purpose

The PFC Analysis Kit for ACQUITY UPLC enables the quantitative analysis of perfluorinated compounds in surface water samples. It is designed to minimize interference from PFCs found in many laboratory instrument components, common HPLC solvents, and laboratory water. The kit contains tubing, fittings, filters, and other components to replace those that can contribute to contamination.The kit also contains a PFC isolator column that helps isolate the background contaminants from the analytes in a sample. The isolator column is in the flow path between the mixer of the binary solvent manager and the injector of the sample manager. During analysis, the isolator column retains the background PFCs, separating them from the analytes of interest. The increased retention time of the background PFCs makes them easily distinguishable by mass analysis.For examples of TIC and MRM chromatograms from the analysis of standard PFC compounds using the PFC analysis kit and method, refer to the application note, ACQUITY UPLC System Solution for Quantifying Trace Levels of Perfluorinated Compounds with an ACQUITY PFC Analysis Kit, on the documentation CD.

PFC Recovery Standard

• Perfluorohexanoic acid (1,2-13C2)

• Perfluoro-n-[1,2-13C2]decanoic acid

• Perfluorohexanesulfonic acid (18O2)

2500 ng for each analyte

2-mL, flame- sealed ampule

Preparation standards for quality control (Continued)

Label Analytes Concentration Vials


Compatible ACQUITY UPLC system configurations

The PFC analysis kit is compatible with an ACQUITY UPLC system composed of the following components:

• Binary solvent manager, with or without i2 Valves • Sample manager• Column heater or high temperature column heater• Sample organizer (optional)

• Tandem quadrupole mass spectrometer (TQ detector, Quattro Premier™ XE, Quattro micro™, or Xevo™ mass spectrometer)

Software control

ACQUITY UPLC systems using the PFC analysis kit require MassLynx™ software version 4.1. Use the QuanLynx™ or TargetLynx™ applications to analyze data. The kit is shipped with a MassLynx project template for running standards and quality control samples to verify system operation. You can also use the project template to run analytical samples. (See “To open the project file” on page 4-3.)

Caution: The kit does not support the ACQUITY UPLC column manager or detectors with Teflon® flow cells (ACQUITY PDA and TUV, for example).

Compatible ACQUITY UPLC system configurations 1-7

2 Setting up the System Hardware

Before installing the PFC analysis kit on an existing system, you must first remove some components. Installing the kit establishes a complete flow path for analyzing PFC-containing samples while minimizing interference from background contamination.

Requirement: Verify that the standard ACQUITY UPLC installation procedure is completed before installing the kit on a new system.See also: The ACQUITY UPLC System Operator’s Guide for basic ACQUITY hardware installation procedures.

Caution: To avoid damaging the kit hardware, use the PFC analysis kit only for applications involving the analysis of perfluorinated compounds.

Contents

Topic PagePreparing an existing system 2-3Preparing the solvent reservoirs 2-5Installing the columns and tubing 2-7Removing the PFC analysis components 2-10

2-1

Required materials

See also: Controlling Contamination in UltraPerformance LC/MS and HPLC/MS Systems (part number 715001307).

Warning: To prevent injury, always wear eye protection and particle-free, powder-free, non-latex gloves when handling tubing that can contain solvents. If possible, drain the tubing before handling.

Locations of required materials

Location Material PFC analysis kit • Flangeless nut extender

• Isolator column • Stainless-steel tubing assemblies,

labeled “Mixer to PFC Isolator” and “Valve to isolator column”

ACQUITY UPLC startup kit Open-end wrenches, 5/8-inch and 1/4-inch

PFC analysis kit or ACQUITY UPLC startup kit

ACQUITY UPLC BEH C18 2.1 μm × 50 mm column

PFC retrofit kit 10-μL sample loop ACQUITY UPLC bottle kit • Bottle-cap inner liners

• Closed-thread bottle caps • Open-thread bottle caps • Bottle-cap-liner plugs • Filters, 10-μm • Clear or amber, glass, borosilicate

bottles: 500-mL, 1000-mL

2-2 Setting up the System Hardware

Preparing an existing system

If you are adding the PFC analysis kit to an existing ACQUITY UPLC system, perform the following procedure before installing the PFC analysis kit components.

To prepare an existing system

1. Using the open-end wrenches, remove the stainless-steel tubing that extends from the mixer outlet of the binary solvent manager to port 5 of the sample manager injection valve. Tips: • Store the tubing for future use.• If you need more room to maneuver inside the binary solvent

manager, you can remove the ends of the pre-installed Tygon® tubing (running from the process waste port) and the PharMed® tubing (running from the needle-clean system waste port) from the pass-through of the binary solvent manager drip tray.

Mixer

Tubing to port 5 of the sample manager injection valve

Tygon and PharMed waste tubing

Tubing from mixer outlet in binary solvent manager

Port 5 of the sample manager injector valve

Preparing an existing system 2-3

2. Using the flangeless-nut extender tool, remove the Teflon solvent tubing from the A1, B1, A2, B2, SNW, WNW, and seal wash (SW) ports of the binary solvent manager.

3. Remove the solvent tubing and solvent bottles from the system, and store them for future use.

4. Use the wrenches to remove the currently installed sample loop. For detailed instructions on how to remove a sample loop from an ACQUITY UPLC system, consult the ACQUITY UPLC System Operator’s Guide.

5. Using the 1/4-inch wrench, remove the tubing that connects the analytical column to the existing detector.

6. Remove from the flow path any detectors employing Teflon flow cells (TUV or PDA, for example). To ensure adequate tubing lengths and distances for the PFC analysis kit components, physically remove these detectors from the ACQUITY stack. Requirements: • Do not install any detectors where flow cells are made of Teflon. • Do not place additional detectors in the ACQUITY stack.


Preparing the solvent reservoirs

To install solvent reservoirs, caps, and tubing

1. In the PFC analysis kit, unwrap the appropriate PEEK™ solvent tubing assemblies for the binary solvent manager’s A1, B1, A2, B2, SNW, WNW, and seal-wash (SW) ports. Tip: Each tubing assembly is labeled with a specific port name.

Recommendation: Match the cap color to the color on the tubing labels and the degasser port.

Caution: To prevent contamination, heed these measures: • Always wear particle-free, powder-free, non-latex gloves when

handling system components. • Select, prepare, and handle solvents and samples correctly. • For best results, use polypropylene vials and caps. • Avoid any vial with a Teflon-lined cap. • Do not wash reservoirs in detergent, alongside other glassware, or

in washing facilities that can retain detergent residue. Store glassware separately from common-use glassware.

• Use clean fittings, tubing, and columns. • Keep laboratory air clean.

Solvent labels and colors

Solvent type Label Color Strong needle wash SNW WhiteWeak needle wash WNW OrangeSolvent A1 A1 YellowSolvent A2 A2 BlueSolvent B1 B1 RedSolvent B2 B2 GreenSeal wash SW Brown

Preparing the solvent reservoirs 2-5

2. Install the bottle cap and solvent filter on one end of the PEEK tubing as follows:

a. Unwind one end of the PEEK tubing, and then slide the outer cap onto it with the threads facing toward the end of the tubing.

b. Slide the end of the tubing through an open hole in the inner liner (but not the center hole).

c. Insert the solvent filter fitting and then the solvent filter onto the end of the tubing.

d. Push the tubing all the way into the solvent filter, and then finger tighten the fitting.

e. If other solvent lines require the same solvent, repeat steps a through d.

f. Insert plugs into the remaining open ports on the inner liner.

3. Install the tubing in the appropriate degasser port as follows:

a. Install the compression screw on the degasser end of the tubing assembly.

b. Slide the lock ring and ferrule onto the tubing, with the metal end nearest the compression screw.

Tip: Finger tighten the compression screw, and then use the flangeless-nut extender tool to tighten it an additional 1/4-turn.

4. If the reservoir has a cap and plastic ring around its neck, remove them.

Inner liner

Center port with 10-μm filter attachedTubing port 5

Label PEEK tubing Inlet filter fitting

Inlet (solvent) filter

Compression screw Lock ring and ferrule


5. Rinse the reservoir with organic solvent, MS-grade water, and, finally, with the intended solvent. Tip: If more aggressive cleaning is required, see “Thorough cleaning of solvent reservoirs” on page 6-4.

6. Fill the reservoir with solvent (see “Preparing eluents and wash solvents” on page 3-3).

7. Insert the solvent filter and tubing in the reservoir. Requirement: Ensure that the entire filter is submerged in the solvent.

8. Fit the inner liner in the outer cap, and tighten the reservoir’s cap.

9. Place the reservoir in the solvent tray.

Installing the columns and tubing

To install the columns and tubing

1. In the PFC analysis kit, locate the two stainless-steel tubing assemblies, labeled “Mixer to PFC Isolator” and “Valve to isolator column”.Requirement: Remove the black, protective O-rings from the stainless-steel tubing before installing them.

2. Using the 5/8-inch and 1/4-inch wrenches, attach one end of the coiled, stainless-steel tubing assembly, labeled “mixer to PFC Isolator”, to the inlet of the PFC Isolator column provided with the PFC analysis kit.

Installing the columns and tubing 2-7

3. Attach the remaining stainless-steel tubing assembly, labeled “Valve to isolator column”, to the PFC Isolator column outlet.

4. Attach the free end of the coiled tubing assembly to the mixer outlet port, and the free end of the “valve to isolator column” tubing assembly to port 5 of the sample manager injector valve. Orient the tubing, as shown below.

Tubing, “Valve to isolator column”

Coiled tubing, “Mixer to PFC Isolator”

PFC Isolator column

Mixer

Port 5

Drip tray

Tubing, “Valve to isolator column”


Flow path through PFC isolator

Requirement: Place the ends of the Tygon tubing (running from the process waste port) and the PharMed tubing (running from the needle-clean system waste port) in the pass-through of the binary solvent manager drip tray, if you removed them to install the kit.

5. Install the 10-μL sample loop (PFC analysis kit). See also: ACQUITY UPLC System Operator’s Guide for detailed instructions on installing a sample loop.

Injection valve port 5

Sample manager

PFC Isolator flow path

Binary solvent manager

Isolator column

Mixer

Installing the columns and tubing 2-9

6. Install the ACQUITY UPLC BEH C18 2.1 × 50 mm column in the column compartment.

7. Using the 1/4-inch wrench, connect the PEEK, “column-to-detector” tubing assembly to the outlet of the analytical column, routing it to a waste container until the flushing procedure is completed (see “Flushing the system” on page 3-2).

Removing the PFC analysis components

You must remove the PFC analysis components to resume analyzing compounds other than PFCs.

To remove the components

1. Disconnect the end of the “valve to isolator column” tubing from the injection valve port 5 in the sample manager.

2. Disconnect the end of the coiled “mixer to PFC isolator” tubing from the mixer in the solvent manager.

3. Remove the PFC Isolator with the stainless steel tubing from the system. Tip: The PEEK tubing can remain.

4. Reinstall the original stainless-steel tubing, connecting the mixer and injector.

5. Check for leaks.

6. Verify system performance. See also: ACQUITY UPLC System Operator’s Guide for detailed instructions on verifying system performance.

7. Store the PFC analysis components for future use.


3 Preparing the System for Analysis

To minimize background PFC contamination, clean the system thoroughly before performing an analysis. Carefully prepare eluents, and wash solvents in the glassware provided with the kit.

Warning: To avoid chemical exposure risk, always observe Good Laboratory Practices when you use this equipment and when you work with solvents and test solutions. Know the chemical and physical properties of the solvents and test solutions you use. See the Material Safety Data Sheet for each solvent and test solution in use.

Caution: To avoid damaging the kit hardware, use the PFC analysis kit only for applications involving the analysis of perfluorinated compounds.

Contents

Topic PageFlushing the system 3-2Preparing eluents and wash solvents 3-3

3-1

Flushing the system

Before performing a PFC analysis, flush the ACQUITY UPLC system thoroughly to remove background contamination that can interfere with the analysis. Flushing is especially important when the system has been used for other types of analyses and it is also required for newly installed systems.

To flush the system

1. After installing the PFC analysis kit components, prepare the solvents:

a. Place all solvent lines except the seal wash in a reservoir containing four liters of 100% MS-grade methanol.

b. Prepare a solution of 10% methanol by mixing 100 mL of methanol with 900 mL of deionized water in a solvent reservoir, and then place the seal wash line in the 10% methanol reservoir.

c. Verify that the column outlet tubing is routed to a waste container.

2. Open the MassLynx application and log on. See also: MassLynx user documentation for instructions on how to complete basic software tasks.

3. On the MassLynx toolbar, click Instrument > MS Console.

4. In the ACQUITY UPLC System tree, click Binary Solvent Manager > Control > Prime seal wash > Yes.Result: Seal-wash priming begins and continues for 15 minutes.

5. In the ACQUITY UPLC System instrument tree, click Sample Manager, and then set the column temperature to 50 °C.

6. In the ACQUITY UPLC System instrument tree, click Binary Solvent Manager.

Caution: • To minimize contamination, use high-purity solvents (MS-grade or

better) and 18-megaohm•cm resistivity water for PFC analysis. • To remove background contamination, flush both existing and newly

installed systems thoroughly before performing a PFC analysis.

3-2 Preparing the System for Analysis

7. In the binary solvent manager window, prime the solvents A1 and B1:

a. Click Control > Prime A/B solvents.

b. Select solvent lines A1 and B1 for 3 minutes, and then click OK.

c. Click Control > Set flow.

d. Enter a flow rate of 0.4 mL/min for 50% A1 and 50% B1, and then click the green check mark.

8. After four hours, repeat step 6 and step 7 for the A2 and B2 solvent lines.

9. While the system is flushing, during step 6 through step 8, prime the syringes.

a. Click Sample Manager > Control > Prime syringes.

b. Select Sample syringe and wash syringes, enter 99 for the number of cycles, and then click OK. Allow approximately three hours.

10. After the syringe priming cycles are completed, wash the needle.

a. Click Control > Wash needle.

b. Enter 500 for both Strong Wash and Weak Wash, and then click OK.

c. Repeat nine times.

11. When the flushing procedure is completed, connect the “Column to detector” tubing (attached to the analytical column outlet) to the mass spectrometer.

12. Run five blank injections before running an analytical sample.

Preparing eluents and wash solvents

Successful analyses with the PFC kit require specific eluents and wash solvents. Use appropriate bottles and closures to minimize contamination and bacterial growth. Carefully prepare the wash solvents and eluents, as described below, in the appropriate glassware.

Caution: Use high-purity solvents (MS-grade or better) and 18-megaohm•cm resistivity water for PFC analysis.

Preparing eluents and wash solvents 3-3

Required materials

• Ammonium acetate• MS-grade methanol• 18 megaohm•cm resistivity water• Solvent reservoir, 1-L, with cap (including plugs for mixing)• Graduated cylinders• Analytical balance

To prepare the solvent for line A1

1. Weigh 0.1542 g of ammonium acetate on an analytical balance, and then transfer it to a 1-L solvent reservoir.

2. Using a graduated cylinder, add 50 mL of methanol to the reservoir.

3. To a clean graduated cylinder, add 950 mL of 18-megaohm•cm resistivity water to the same reservoir.

4. Cap the bottle, and plug any holes, to prevent leaks.

5. Shake the solvent bottle to mix the contents.

Wash solvents for PFC analyses on the ACQUITY UPLC system

Solvent line SolventStrong needle wash 1 L of 100% methanolWeak needle wash 1 L of 25% methanol in waterSeal wash 1 L of 10:90 methanol/water (use 5-minute washes)

Eluents for PFC analyses on the ACQUITY UPLC system

Solvent line EluentA1 1 L of 2 mM ammonium acetate in 5:95 methanol/water (see

“To prepare the solvent for line A1”, below)B1 1 L of 100% methanolA2 Same as B1 (can place solvent line in B1 reservoir)B2 Same as B1 (can place solvent line in B1 reservoir)

3-4 Preparing the System for Analysis

4 Verifying System Performance

By running a standard mixture of perfluorinated compounds on a system equipped with the PFC analysis kit, you verify that the system is performing correctly. This chapter assumes you are using the Waters TQ Detector mass spectrometer. See also: The mass spectrometer’s user documentation for specific information on using it with the PFC analysis kit.

Warning: To avoid chemical exposure risk, always observe Good Laboratory Practices when you use this equipment and when you work with solvents and test solutions. Know the chemical and physical properties of the solvents and test solutions you use. See the Material Safety Data Sheet for each solvent and test solution in use.

Contents

Topic PagePreparing the standards 4-2Performing the verification test 4-2Interpreting the test data 4-9

4-1

Preparing the standards

Required materials

• PFC calibration standards from the analysis kit• Pipetter with polypropylene tips capable of delivering 10 to 1000 μL• 18-megaohm•cm resistivity water• MS-grade methanol• Vials, 12 × 32-mm, 2-mL, polypropylene, screw-neck, with polyethylene,

septumless caps

To prepare the verification standard and blanks

1. Using a pipetter, transfer 1 mL of PFC calibration standard #1 to a mixing vial.

2. Deliver 3 mL of deionized water to the vial, so that the concentration of the resulting mixture is 1.25 ng/mL. Label this vial “Test solution #1”.

3. Prepare a 25% methanol solution by mixing 750 mL deionized water with 250 mL methanol.

4. Transfer test solution #1 to an individual UPLC sample vial for analysis as a verification standard.

5. Transfer the 25% methanol solution to UPLC sample vials to use as analysis blanks.

Performing the verification test

Requirement: Before you run a test to verify whether the system is operating properly for PFC analysis, ensure that the system is properly flushed (see page 3-2).The documentation CD shipped with the system includes a predefined project file for the verification test. Copy the project folder to the MassLynx directory on the hard drive of your computer before opening the project.The method in the predefined project focuses on ten, common, perfluorinated compounds. The calibration standards provided in the kit contain six additional analytes (see page 1-2) that a modified method can include.

4-2 Verifying System Performance

Opening the project fileThe PFC_Analysis.PRO project folder contains the sample list PFC_analysis.SPL.

To open the project file

1. From the system CD, copy the PFC_Analysis.PRO folder to the MassLynx directory on the hard drive (usually C:\MassLynx).

2. Ensure the ACQUITY UPLC system is powered-on.

3. Open the MassLynx application and log on.See also: MassLynx user documentation for instructions on how to perform basic software tasks.

4. Click File > Open Project, and select the project folder “PFC_Analysis.PRO”.Result: The project opens and the sample list PFC_analysis.SPL appears. The list contains representative test samples and data for reference.

5. Click File > Open.

6. Select the PFC_analysis_run sample list to run samples.

Calibrating and tuning the mass spectrometer

Through the MassLynx IntelliStart™ interface, you can run an appropriate reference solution for ESI-negative analysis to calibrate and tune the mass spectrometer, which involves these tasks:

• Preparing the instrument• Setting up the Tune window and preparing the mass spectrometer’s

solvent delivery system• Generating a calibration curve using a reference solution• Tuning the system on the reference material

Tips: • For information about using IntelliStart, consult the MassLynx online

Help and the Waters TQ Detector Operator’s Guide. • Quattro Micro and Quattro Premier mass spectrometers require tuning

and calibration.

Performing the verification test 4-3

To calibrate the TQ detector and edit the tune file for PFC analysis

1. In the MS Console, launch the ACQUITY UPLC Console.

2. In the system tree, select TQD, and then click IntelliStart.

3. In the IntelliStart window, click Operate, API gas, and Tune.

4. In the Tune window, click File > Open.

5. In C:/MassLynx/PFC_Analysis.PRO/ACQUDB, select the tune file Instrument.ipr, and then click Open. Result: The Tune window displays the tune values.

6. Place the bottle of API calibration solution in reservoir A.

7. On the Fluidics tab, select reservoir A, and then click Purge fluidics system.

8. Select infusion as the flow state, and then specify 10 µL/min.

9. Click to start, and then minimize the Tune window.

10. In the IntelliStart window, select Instrument Setup, and then click Start.Result: The IntelliStart Setup Parameters window opens.

11. In the IntelliStart Setup Parameters window’s Instrument setup tab, enter or verify the following parameters:

Instrument setup conditions

Parameter Value Instrument tune masses 126.90, 426.69, 876.38, and 1625.85 Instrument tune file Browse to

C:/MassLynx/PFC_Analysis.PRO/ ACQUDB/Instrument.ipr

Target resolution MS and MSMS resolution: 0.85 Da Calibration range Low mass: 2.00 Da

High mass: 2040.00 Da Slow speed: 6.00 s Fast speed: 0.20 s


12. Click the Sample tune tab, and enter or verify the following parameters:

13. To start calibration, click the green check mark.

14. When calibration is complete, maximize the Tune window.

15. Enter these values in the Tune window:

Calibration files Reference: NainegCalibration: PFC_[date].cal

Calibration reporting Print calibration report selectedFluidics Enabled; reservoir A

Sample tune conditions

Parameter Value Sample tune masses 126.90, 426.69, 876.38, and 1625.85 Sample tune file Browse to

C:/MassLynx/PFC_Analysis.PRO/ ACQUDB/PFC_analysis.ipr

Fluidics Enabled; Reservoir A; Flow state: Infusion

Tune window conditions

Parameter ValueIonization mode ESI negativeCapillary voltage −3.6 kVCone voltage As is Extractor −3 VSource temperature 150 °CDesolvation temperature 400 °CDesolvation gas flow 800 L/hrCone gas flow 0 L/hrMode MSMS

Instrument setup conditions (Continued)

Parameter Value


16. Save the tune file as “PFC_analysis.ipr”.

To prepare the chromatography method and run blank and PFC standard samples

1. Place the blank sample vial in the sample tray position 2:1 and the test solution #1 (1.25 ng/mL PFC standard) sample vial in tray position 2:2.

2. In the MassLynx PFC_analysis_run.spl sample list, open the MS file named PFC_10MRM_9min.

3. Verify that the values match those in the following table:

Collision gas Argon, at approximately 3.2 × 10−4 kPa (3.2 × 10−3 mBar, 4.6 × 10−5 psi); 0.1 mL/min flow rate

Acquisition mode MRM

Settings for MRM of 10 mass pairs page

Parameter

Value

Parent (m/z)

Daughter (m/z)

Dwell (Secs)

Cone (Volts)

Coll Energy (eV)

Total run time 9.00 min Channels 299.00 80.00 0.020 50.00 23.00

313.00 269.00 0.020 16.00 7.00 363.00 319.00 0.020 15.00 7.00399.00 80.00 0.020 60.00 29.00 413.00 369.00 0.020 16.00 7.00 463.00 419.00 0.020 20.00 8.00 499.00 80.00 0.020 65.00 33.00 513.00 469.00 0.020 22.00 8.00 563.10 519.10 0.020 22.00 8.00 613.10 569.10 0.020 22.00 8.00

Tune window conditions (Continued)

Parameter Value


4. In the MassLynx project inlet file (PFC_inlet_9min), verify that the binary solvent manager and sample manager instrument methods match the values in the following table:

Ionization mode ESI negative Repeats 1 Span 0.2 Use tune cone settings

Not selected

Use tune coll energy

Not selected

Retention window Start: 0; End: 9

UPLC conditions

Parameter ValueWeak wash 25% methanol in water (500 μL)Strong wash Methanol (500 μL)Seal wash 90:10 water/methanol (5 minutes)Column temperature 50.0 °CSample temperature 20.0 °CSample loop and injection volume

10 μL (Full Loop)

Column ACQUITY UPLC BEH C18 2.1 × 50 mmMobile phase A1 2 mM ammonium acetate in 95:5

water/methanolMobile phase B1 100% methanolRun time 9.00 min

Settings for MRM of 10 mass pairs page (Continued)

Parameter

Value

Parent (m/z)

Daughter (m/z)

Dwell (Secs)

Cone (Volts)

Coll Energy (eV)


5. Confirm that you added 5 blank samples and your verification standard samples to the sample queue. Tip: To add a new test sample to the sample queue, click Samples > Add, and then specify the appropriate sample information. Use the optimized, mass-spectrometer information. Recommendation: At system startup, always run 5 blank samples before your analysis, to equilibrate the system and monitor the level of background PFC contamination.

6. Confirm the following selections:• MS tune file is PFC_analysis. • Injection volume in the sample list is 10 μL. • Mobile phases match the solvents in the table titled “Gradient

method” on page 4-8.

7. Prime the A and B pumps for 3 min.

Low pressure limit 0 High pressure limit 15000 psi

Gradient method

Time (min) Flow rate (mL/min) %A %B Curve

0 0.400 75.0 25.00.50 0.400 75.0 25.0 65.00 0.400 15.0 85.0 65.10 0.400 0.0 100.0 65.60 0.400 0.0 100.0 67.00 0.550 0.0 100.0 19.00 0.400 75.0 25.0 112.00 0.000 75.0 25.0 11

UPLC conditions (Continued)

Parameter Value


8. Start the flow at 0.4 mL/min with 75% solvent A for 3 min, and inspect all connections and columns for leaks.

9. In the MassLynx sample list window, click Run. Recommendations

• Set up a refined MS method, with multiple MRM time windows, allowing more time to scan target MRM transitions at the target PFC peak, which yields better signal-to-noise ratios and peak detection. (See the application note in the startup CD, “Quantifying Trace Levels of Perfluorinated Compounds with an ACQUITY PFC Analysis Kit”.)

• To achieve optimal sensitivity for each PFC analyte, use IntelliStart’s Develop method functions, which fine-tune the MRM transitions, cone voltage, and collision energy for each perfluorinated compound. (See the MassLynx online Help and the Waters TQ Detector Operator’s Guide.)

• At system startup, and before each run, monitor the level of background PFC contamination, and verify the performance of the isolator column by running 5 blank injections and a PFC standard.

Interpreting the test data

The system is performing correctly when the following statements about your processed data are true:

• The retention time difference between the perfluorooctanoic acid (PFOA) analyte peak and the leading PFOA contaminant peak is at least 0.4 minutes.

• The PFOA and perfluorononanoic acid (PFNA) peaks are baseline-resolved.

To process data from the verification standard

1. In the MassLynx window, click the line number of the standard for which you are processing data.

2. In the sample list menu bar, click Chromatogram.Result: The TIC chromatogram for the standard appears.

Interpreting the test data 4-9

TIC chromatogram of PFC standard (ESI negative mode)

3. In the Chromatogram window, click Display > Mass, and then double-click the mass transition for PFOA (413 > 369) in the function list.

4. Select Add Trace (or Replace trace) > OK.Result: The chromatogram for PFOA appears.

5. Click Process > Smooth, and then enter 3 for the window size and 2 for the number of smoothing operations.

6. Select Mean > OK.Result: The smoothed chromatogram for PFOA appears.Tip: To delete the TIC and the unsmoothed chromatogram, click within the plot area (a colored square appears to the left of the plot), press the keyboard’s Delete button, and then click OK.

7. Click Display > View.


8. In the Chromatogram Display View window, select Dot from the Grid drop-down menu, and click OK.

Smoothed PFOA chromatogram

9. Expand the view of the analyte peak and the leading contaminant peaks.Tip: To draw a box around the peaks of interest, click the top-left corner of the region of interest, and then drag diagonally downward to the axis, without extending the box below the axis.

10. Measure the distance between the apex of the PFOA peak and the half-height of the first contaminant peak.Tip: Click and drag a horizontal line between the two points. The retention time difference between the two points appears in the bottom-left corner of the chromatogram window.


Measuring the difference in retention time

11. Verify that the difference in retention time is at least 0.4 minutes.Tip: A distance of less than 0.4 minutes can indicate a problem (see “Potential analysis problems and probable solutions” on page 6-2).

12. In the Chromatogram window, click Display > Mass, and then double-click the mass transition for PFNA in the Function list (463 > 419).

13. Select Add Trace > OK.Result: The chromatogram for PFNA appears.

14. Click Process > Smooth, and then specify 3 as the window size and 2 as the number of smooths.

15. Select Mean > OK.Result: The smoothed chromatogram for PFNA appears.Tip: To delete the unsmoothed PFNA chromatogram, click within the plot area (a colored square appears to the left-hand side of the plot), and then press Delete > OK.

16. Click Display > View.

17. In the Chromatogram Display View window, select Overlay Graphs > OK.


PFOA and PFNA peaks

18. Examine the PFOA and PFNA peaks in the plots that you overlaid, and verify they are baseline-resolved. Tip: Peaks not baseline-resolved can indicate a problem (see “Potential analysis problems and probable solutions” on page 6-2).


5 Preparing Samples

The sample preparation procedure in this chapter serves as a guideline for preparing a surface water sample for PFC analysis.

Typical sample preparation procedure

Required materials

• Oasis HLB PLUS extraction cartridge, 225 mg, 60 μm• SPE extraction manifold • MS-grade methanol• 18-megaohm•cm resistivity water• PFC Internal Standard from the analysis kit, 1-ng/mL dilution

(page 1-5)• Surface water sample• Nitrogen gas supply

Caution: • To avoid sorption of PFC analytes on the walls of containers, use

only polypropylene sample vials. • To limit contamination, wash lab ware with nonphosphate, alkaline

soap, and rinse it well with warm water. Rinse with methanol prior to use. If previous use is unknown, see “Thorough cleaning of solvent reservoirs” on page 6-4.

Typical sample preparation procedure 5-1

Requirement: The system requires a supply of dry, oil-free nitrogen with a purity of at least 95%. The use of nitrogen cylinders is acceptable.

• Centrifuge tube, 15-mL polypropylene• Pipetter with polypropylene tips capable of delivering 10 to 1000 μL• Nitrogen evaporator or nitrogen evaporator with a heated water-bath

evaporator capable of attaining 25 to 60 °C and 69 to 103 kPa (0.69 to 1.03 bar, 10 to 15 psi) nitrogen.

• Sample vials, 12 × 32-mm, 2-mL, polypropylene, screw-neck, with polyethylene, septumless caps

• Vortex mixer• Polypropylene bottle with cap

To prepare a sample for PFC analysis

1. Set up the SPE extraction manifold following the manufacturer’s recommendations.

2. Place Oasis HLB SPE cartridges onto the extraction manifold.

3. At a flow rate of less than 10 mL/min, pass 10 mL of methanol through the Oasis cartridge to condition it.

4. Pass 10 mL of 18-megaohm•cm resistivity water through the cartridge.

5. Measure 500 mL of water sample into the polypropylene bottle.

6. Using a pipetter, add 50 μL of 1-ng/mL internal standard solution to the sample.

7. At a flow rate of 5 mL/min, pass 500 mL of the water sample (including the internal standard) through the Oasis cartridge.

8. To purge the cartridge and remove water, pass nitrogen through it until the cartridge returns to its original (dry) color, from top to bottom (approximately 20 minutes).

Caution: If copper tubing is used for the nitrogen line, the copper must be chemically cleaned; if stainless steel tubing is used, the stainless steel must be medical grade. Ensure that there are no soldered or brazed joints in the line, as these may result in contamination of the instrument with tin or lead oxide. Any joints in the nitrogen line must be compression fittings.

5-2 Preparing Samples

9. At a flow rate of 1 mL/min, pass 2 mL of methanol through the cartridge, and collect the solvent in a 15-mL polypropylene centrifuge tube.

10. Using a nitrogen/bath evaporator set to 35 °C, evaporate the eluent in the tube until it reaches a total volume of 500 μL.

11. Using a pipetter, deliver 300 μL of the methanol eluent to a UPLC sample vial.

12. Deliver 900 μL of 18-megaohm•cm resistivity water to the same vial.Result: The target sample mixture is 1:3 eluent/water.

13. Using a vortex mixer, mix the vial for 5 seconds or until the sample is uniform.

Typical sample preparation procedure 5-3

5-4 Preparing Samples

6 Troubleshooting

See page 6-2 for information about how to solve potential analysis problems. See also: • ACQUITY UPLC System Operator’s Guide for general information

on troubleshooting the system. • Controlling Contamination in UltraPerformance LC/MS and

HPLC/MS Systems (part number 715001307).

Contents

Topic PagePFC analysis troubleshooting 6-2Thorough cleaning of solvent reservoirs 6-4

6-1

PFC analysis troubleshooting

Consult the table below for help solving problems with your analysis.

Potential analysis problems and probable solutions

Symptom Possible cause SolutionContaminant peaks are larger than expected and are comparable to analyte peaks

System has not been sufficiently flushed

Repeat the methanol flushing procedure (page 3-2) until the background contamination is at a reasonable level.

Retention time difference between PFOA (analyte) peak and leading PFOA-contaminant peak is less than 0.4 minutes; analyte peak shapes are normal

Incorrect flow rate Verify the method. Adjust the flow rate to the correct value.

Incorrect solvent composition

Verify the method and the solvent supply. Adjust the solvent composition as needed.

PFC Isolator column is not performing correctly

Replace the PFC Isolator column.

Retention time difference between PFOA (analyte) peak and leading PFOA-contaminant peak is less than 0.4 minutes; noticeable broadening of peaks

Analytical column is not performing correctly

Compare current resolution of PFOA and PFNA peaks to the resolution at installation. Verify column efficiency. Replace column if needed.

6-2 Troubleshooting

PFOA and PFNA peaks are not baseline-resolved

Incorrect flow rate Verify the method. Adjust the flow rate to the correct value.

Incorrect solvent composition

Verify the method and the solvent supply. Adjust the solvent composition as needed.

Analytical column is not performing correctly

Compare current resolution of PFOA and PFNA peaks to the resolution at installation. Verify column efficiency. Replace column if needed.

PFC peaks appear in blank runs along with delayed background contaminant peaks

PFC contamination in injector

Repeat the flushing procedures specific to the sample manager (page 3-3). Prime the syringes and wash the needle until the peaks no longer appear.

PFC contamination in sample vial

Obtain clean sample vials. For best results, use polypropylene vials and caps. Avoid any vial with a Teflon-lined cap.

PFC contamination in solvents

Use higher purity solvents for your analysis. See the Cautions, below.

Caution: To avoid contamination, use MS-grade solvents and 18-megaohm•cm resistivity water for your analysis. Purify the solvents by solid-phase extraction (Waters Oasis MAX or HLB cartridges, for example), if necessary.

Potential analysis problems and probable solutions (Continued)

Symptom Possible cause Solution

PFC analysis troubleshooting 6-3

Thorough cleaning of solvent reservoirs

When a container’s history is unknown, you must clean it thoroughly.

To thoroughly clean solvent reservoirs

1. Sonicate with MS-quality 10% formic or nitric acid.

2. Sonicate with water.

3. Sonicate with methanol.

4. Sonicate with water.

5. Repeat steps 1 through 4, twice.

6. Rinse with organic solvent, MS-grade water, and, finally, with the solvent that the reservoir will contain.

Recommendations: • Do not wash reservoirs in detergent, alongside other glassware, or in

washing facilities that can retain detergent residue. • Store glassware separately from common-use glassware.

To clean solvent reservoirs contaminated with microbial growth

1. Treat the reservoirs in an autoclave.

2. Remove and replace all filters and tubing between the solvent reservoir and the instrument.

3. Purge the system with methanol, and let it stand overnight.

Caution: Replacing check valves can increase the background PFC contamination in the system. To avoid additional contamination, flush the system thoroughly after replacing check valves (page 3-2).

6-4 Troubleshooting

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.

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 warningsWarning 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.

Contents

Topic PageWarning symbols A-1Caution symbol A-5Warnings that apply to all Waters instruments A-5Electrical and handling symbols A-12

Warning symbols A-1

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.)

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

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

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.

Mass spectrometer shock hazard

This warning applies to all Waters mass spectrometers.

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 690 kPa (6.9 bar, 100 psi) during an analysis in which flammable solvents are used. Also ensure a gas-fail connection is connected to the LC system so that the LC solvent flow stops if the nitrogen supply fails.

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

Warning symbols A-3

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.

Chemical hazard warning

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

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 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).

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 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 equipment guidelines in this section.

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: qualsiasi modifica o alterazione apportata a questa unità e non espressamente autorizzata dai responsabili per la conformità fa decadere il diritto all'utilizzo dell'apparecchiatura da parte dell'utente.

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.

注意：未經有關法規認證部門允許對本設備進行的改變或修改,可能會使使用者喪失操作該設

備的權利。

注意：未经有关法规认证部门明确允许对本设备进行的改变或改装，可能会使使用者丧失操作该设备的合法性。

주의: 규정 준수를 책임지는 당사자의 명백한 승인 없이 이 장치를 개조 또는 변경할 경우, 이 장치를 운용할 수 있는 사용자 권한의 효력을 상실할 수 있습니다.

注意：規制機関から明確な承認を受けずに本装置の変更や改造を行うと、本装置のユーザーとしての承認が無効になる可能性があります。


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.

Warnings that apply to all Waters instruments A-7

Attenzione: fare attenzione quando si utilizzano tubi in materiale polimerico sotto pressione:• Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero

pressurizzati.• Spegnere tutte le fiamme vive nell'ambiente circostante.• Non utilizzare tubi eccessivamente logorati o piegati.• Non utilizzare tubi non metallici con tetraidrofurano (THF) o acido solforico

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

rigonfiamenti nei tubi non metallici, riducendo notevolmente la pressione di 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.

警告：當在有壓力的情況下使用聚合物管線時，小心注意以下幾點。

• 當接近有壓力的聚合物管線時一定要戴防護眼鏡。

• 熄滅附近所有的火焰。

• 不要使用已經被壓癟或嚴重彎曲管線。

• 不要在非金屬管線中使用四氫呋喃或濃硝酸或濃硫酸。

• 要了解使用二氯甲烷及二甲基亞楓會導致非金屬管線膨脹，大大降低管線的耐壓能力。


警告：当有压力的情况下使用管线时，小心注意以下几点：

• 当接近有压力的聚合物管线时一定要戴防护眼镜。

• 熄灭附近所有的火焰。

• 不要使用已经被压瘪或严重弯曲的管线。

• 不要在非金属管线中使用四氢呋喃或浓硝酸或浓硫酸。

• 要了解使用二氯甲烷及二甲基亚枫会导致非金属管线膨胀，大大降低管线的耐压能力。

경고: 가압 폴리머 튜브로 작업할 경우에는 주의하십시오.• 가압 폴리머 튜브 근처에서는 항상 보호 안경을 착용하십시오.• 근처의 화기를 모두 끄십시오.• 심하게 변형되거나 꼬인 튜브는 사용하지 마십시오.• 비금속(Nonmetallic) 튜브를 테트라히드로푸란(Tetrahydrofuran: THF) 또는 농축 질산 또는 황산과 함께 사용하지 마십시오.

• 염화 메틸렌(Methylene chloride) 및 디메틸술폭시드(Dimethyl sulfoxide)는 비금속 튜브를 부풀려 튜브의 파열 압력을 크게 감소시킬 수 있으므로 유의하십시오.

警告：圧力のかかったポリマーチューブを扱うときは、注意してください。

• 加圧されたポリマーチューブの付近では、必ず保護メガネを着用してください。

• 近くにある火を消してください。

• 著しく変形した、または折れ曲がったチューブは使用しないでください。

• 非金属チューブには、テトラヒドロフラン(THF)や高濃度の硝酸または硫酸などを流

さないでください。

• 塩化メチレンやジメチルスルホキシドは、非金属チューブの膨張を引き起こす場合が

あり、その場合、チューブは極めて低い圧力で破裂します。


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 die eingebauten Sicherheitseinrichtungen unter Umständen nicht ordnungsgemäß funktionieren.

Attenzione: si rende noto all'utente che l'eventuale utilizzo dell'apparecchiatura secondo modalità non previste dal produttore può compromettere la protezione offerta dall'apparecchiatura.

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.

警告：使用者必須非常清楚如果設備不是按照製造廠商指定的方式使用，那麼該設備所提供的保護將被消弱。

警告：使用者必须非常清楚如果设备不是按照制造厂商指定的方式使用，那么该设备所提供的保护将被削弱。

경고: 제조업체가 명시하지 않은 방식으로 장비를 사용할 경우 장비가 제공하는 보호 수단이 제대로 작동하지 않을 수 있다는 점을 사용자에게 반드시 인식시켜야 합니다.

警告：ユーザーは、製造元により指定されていない方法で機器を使用すると、機器が提供している保証が無効になる可能性があることに注意して下さい。


Warning: To protect against fire, replace fuses with those of the type and rating printed on panels adjacent to instrument fuse covers.Attention: pour éviter tout risque d'incendie, remplacez toujours les fusibles par d'autres du type et de la puissance indiqués sur le panneau à proximité du couvercle de la boite à fusible de l'instrument.Vorsicht: Zum Schutz gegen Feuer die Sicherungen nur mit Sicherungen ersetzen, deren Typ und Nennwert auf den Tafeln neben den Sicherungsabdeckungen des Geräts gedruckt sind.Attenzione: per garantire protezione contro gli incendi, sostituire i fusibili con altri dello stesso tipo aventi le caratteristiche indicate sui pannelli adiacenti alla copertura fusibili dello strumento.Advertencia: Para evitar incendios, sustituir los fusibles por aquellos del tipo y características impresos en los paneles adyacentes a las cubiertas de los fusibles del instrumento.

警告 : 為了避免火災，更換保險絲時，請使用與儀器保險絲蓋旁面板上所印刷之相同類型與規格的保險絲。

警告 : 为了避免火灾，应更换与仪器保险丝盖旁边面板上印刷的类型和规格相同的保险丝。

경고: 화재의 위험을 막으려면 기기 퓨즈 커버에 가까운 패널에 인쇄된 것과 동일한 타입 및 정격의 제품으로 퓨즈를 교체하십시오.

警告 : 火災予防のために、ヒューズ交換では機器ヒューズカバー脇のパネルに記載されているタイプおよび定格のヒューズをご使用ください。


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.


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!

Electrical and handling symbols A-13

Index

Aadding samples to the sample queue

4-8analysis blanks 4-2, 4-8analysis method 4-2analytes not included in method 4-2analytical column 1-2application description 1-6

Bbaseline resolution of PFOA and PFNA

4-9, 4-13biohazard warning A-4blank runs 4-2, 4-8bottles, for solvents 3-3burst warning A-3

Ccalibrating and tuning 4-3, 4-4calibration standard solutions 1-3caution symbol A-5check valves, replacing 6-4chemical hazard warning A-4chromatograms of standards 1-6, 4-10,

4-11, 4-12, 4-13chromatography method 4-6cleaning the system 2-10, 3-1, 3-2column manager, using with kit 1-7column, for PFC analysis 1-2contamination 2-2, 6-1

Ddata processing 4-9detectors compatible with kit 1-7, 4-1

EEC Authorized Representative vielectrical symbols A-12

eluents 3-3equipment guidelines A-5ESI-negative analysis 4-3existing system, using with kit 2-3

Fflammable solvents A-3flow path 1-6flushing the system 2-10, 3-1, 3-2

Ggradient method 4-6guidelines for preparing a sample 5-1

Hhandling symbols A-13HLB cartridges 1-2, 5-1

Iinlet method 4-6IntelliStart calibrating and tuning 4-3,

4-4intended use vinternal standard 1-5, 5-1ISM classification visolator column 1-3

Kkit

application verification 4-1contents 1-2hardware 1-3, 2-1usage guidelines iv

LLC method 4-6

Index-1

Mmass spectrometer

calibrating 4-4compatibility with kit 1-7, 4-1

mass spectrometer shock hazard A-3mass transitions for PFCs 4-10, 4-12MassLynx IntelliStart 4-3, 4-4method 4-2

chromatography 4-6mass spectrometer calibration 4-4

methodology 1-6multiple detectors 2-4

Ppass criteria for verification test 4-9PDA detector, using with kit 1-7, 2-4perfluorinated compounds. See PFCsPFC analysis kit 1-2, 1-3, 2-1

chromatograms 1-6, 4-10, 4-11, 4-12, 4-13

contamination 2-2, 6-1eluents and wash solvents 3-3flow path 1-6flushing the system 2-10, 3-1, 3-2installation 2-1isolator column 1-3methodology 1-6preparing samples 5-1project template 1-7quality control standards 1-5running a standard 4-1standard solutions 1-3

preparing 4-2system configurations 1-7troubleshooting 6-1usage guidelines ivusing with an existing system 2-3

PFC Isolator column 1-3, 1-6

PFCsin standards 1-4, 4-2using kit for analysis of compounds

1-6PFNA mass transition 4-12PFOA

mass transition 4-10retention time difference 4-9

PFOA and PFNA peak resolution 4-9, 4-13

preparation standards 1-5preparing

eluents and wash solvents 3-3samples 5-1standards 4-2

processing data 4-9project template 1-7, 4-2purifying solvents 6-3

Qquality control standards 1-5QuanLynx, analyzing data 1-7Quattro micro mass spectrometer 1-7,

4-1Quattro Premier XE mass

spectrometer 1-7, 4-1

Rrecovery standard 1-5, 1-6replacing check valves 6-4resolution of PFOA and PFNA peaks

4-9, 4-13resuming non-PFC analysis 2-10retention time difference between

PFOA peaks 4-9retrofit kit 1-3running a standard 4-1

Index-2

Ssafety advisories A-1sample

preparation 5-1queue setup 4-8vials 1-2

sample loop 1-3, 2-9setting up ESI-negative analysis 4-3software control with MassLynx 1-7solid phase extraction

cartridges 1-2, 5-1solvent

grades 3-2, 3-3reservoirs 3-3tubing assemblies 2-5

stainless-steel tubing 2-2, 2-7standard solutions 1-3, 4-2

analytes 1-4internal and recovery standards

1-5, 5-1running a standard 4-1

symbolscaution A-5electrical A-12handling A-13warning A-1

system configurations compatible with kit 1-7

system verification 4-1

TTargetLynx, analyzing data 1-7TQ Detector mass spectrometer 1-7,

4-1troubleshooting 6-1tubing assemblies 2-2, 2-5, 2-7tune file 4-4tuning and calibrating 4-3, 4-4TUV detector, using with kit 1-7, 2-4

I

Uusage guidelines iv

Vverification test 4-1, 4-2

criteria 4-9method 4-2

vials for sample analysis 1-2

Wwarning symbols A-1, A-5wash solvents 3-3

XXevo mass spectrometer 1-7, 4-1

Index-3

Index-4

pfc analysis kit for acquity uplc system guide€¦ · performance. safety advisories consult...

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