SCIEX Anion Analysis Kit Handbuch

SCIEX Anion Analysis Kit

For P/ACE™ MDQ and P/ACE™ MDQ plus Capillary

Electrophoresis Systems

Instruction Guide

A49108AE

May 2015

AB Sciex

500 Old Connecticut Path

Framingham, MA 01701

USA

AB Sciex Pte. Ltd and its affiliates disclaims all warranties with respect to this document,

expressed or implied, including but not limited to those of merchantability or fitness for a

particular purpose. In no event shall AB Sciex Pte. Ltd. and its affiliates be liable, whether in

contract, tort, warranty, or under any statute or on any other basis for special, incidental, indirect,

punitive, multiple or consequential damages in connection with or arising from this document,

including but not limited to the use thereof.

For research use only. Not for use in diagnostic procedures.

The trademarks mentioned herein are the property of AB Sciex Pte. Ltd. or their respective

owners.

AB SCIEX™ is being used under license and SCIEX Separations is a part of AB Sciex.

Find us on the World Wide Web at www.sciex.com.

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Contents

Chapter 1 Using the Anion Analysis Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5

Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5

Materials and Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6

Storing Kit Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6

Cleaning Vial Caps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7

Turning on the UV Lamp . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7

Cleaning the Capillary Interface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7

Installing the Capillary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7

Conditioning a New Capillary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8

Storing the Capillary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8

Preparing the Buffer Trays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8

Preparing the Sample . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9

PCR Vial Setup–P/ACE MDQ System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10

Universal Vial Setup–P/ACE MDQ plus System . . . . . . . . . . . . . . . . . . . . . . .11

Running Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11

Initial Conditions for All Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12

Time Programs for Anion Separation Methods . . . . . . . . . . . . . . . . . . . . . . . .13

Time Program for Anion Shutdown Method . . . . . . . . . . . . . . . . . . . . . . . . . .14

Checking System Performance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14

Using the Anion Organic Test Mix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14

Using the Anion Inorganic Test Mix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15

Integration Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15

Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16

Appendix A Filter Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17

Installing the 230 nm Filter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17

Appendix B System Configuration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19

Configuring the P/ACE™ MDQ or P/ACE™ MDQ plus System . . . . . . . . . . . . .19

Activating Caesar Integration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .21

Contents

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Using the Anion Analysis Kit

Introduction

The SCIEX Anion Analysis Kit contains the supplies necessary for the separation and

quantitation of anions, using the SCIEX P/ACE MDQ and P/ACE MDQ plus Capillary

Electrophoresis Systems. Each anion kit yields approximately 500 tests.

This kit permits the analysis of small inorganic anions and organic acids, which are often UV

transparent. For this reason, the separation buffer contains a chromophore, and detection is

achieved in indirect mode.

The separation method is performed under reverse polarity so that the negatively charged ions

migrate toward the anode (the positively-charged electrode). In addition, the capillary is

dynamically coated with a polycation, which reverses the electro-osmotic flow (EOF) toward the

anode, thus reducing the separation time while maximizing migration time reproducibility.

IMPORTANT: The main focus of this application is in the biopharmaceutical market. This product

can also be used for environmental testing and food and beverage markets. This product is for

research use only. It is not for use in diagnostic procedures. No clinical decision or patient

notification may be made based on results using this research assay.

Safety

Refer to the Safety Data Sheets (SDS) information, available at sciex.com/safety-data-sheets,

regarding the proper handling of materials and reagents. Always follow standard laboratory

safety guidelines.

Note: The system must be equipped with a UV detector and a 230 nm filter to perform

this assay.

Using the Anion Analysis Kit

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Materials and Reagents

Storing Kit Components

Upon receipt, store all components at room temperature and away from direct sunlight.

Table 1-1 Kit Contents (PN A53537)

Component Quantity

Anion Coating 1

Anion Separation Buffer 1

Conditioner — Na 1

Anion Acid Rinse 1

Anion Internal Standard 1

Anion Organic Test Mix 1

Anion Inorganic Test Mix 1

Capillary, 50 cm, 75 µm I.D. 3 pieces

Rinse Solution 1

Ion Analysis Insert 1

Table 1-2 Materials Required but Not Included in This Kit

Description Part

Number

P/ACE System

MDQ MDQ plus

230 nm filter (see note below) 144433 

Adequate pipettes and pipette tips 

PCR vials (100-pack) 144709 

2 mL glass vials (100-pack) 144980 

Red caps for 2 mL glass vials (100-pack) 144648 

PCR vial holders (50-pack) 144657 

PCR vial springs (10-pack) 358821 

Gray caps for PCR vials (50-pack) 144656 

Universal plastic vials (100-pack) A62251 

Blue rubber caps for universal vials (100-pack) A62250 

Note: A 230 nm filter is required for the analysis. This filter is included with the P/ACE

MDQ plus system and is installed at position 4. However, it is not provided for the P/ACE

MDQ system and must be purchased separately.

Using the Anion Analysis Kit

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Cleaning Vial Caps

1. Using a clean beaker, rinse the vial caps twice with double-deionized (DDI) water.

Do not use soap.

2. Let the caps soak in DDI water for at least one hour, making sure the caps are

completely submerged.

3. Remove the caps from the water.

4. To dry the caps, either place them in an oven set at 55°C for two hours, or allow

them to dry overnight at room temperature covered by clean, lint-free laboratory

tissue.

The vial caps become compressed and lose elasticity during use, which can lead to

pressure failures and current leakage errors. Therefore, reusing caps is not

recommended.

Turning on the UV Lamp

Turn on the UV lamp and allow the system to warm up for at least 30 minutes prior to

experimentation.

Cleaning the Capillary Interface

Carefully clean the system electrodes and interface block as described in the “Maintenance

Procedure” section of the instrument manual. Repeat this procedure after every 24 hours of

operation.

Installing the Capillary

1. Install a 75 µm I.D., 60.2 cm long (50 cm from injection site to detector) fused-silica

capillary into a capillary cartridge using the Capillary Cartridge Rebuild Instructions

(PN 144655).

2. Use an 800 µm aperture in the cartridge. This aperture is labeled with an “8.”

3. After the capillary has been installed in the cartridge, insert the cartridge in the

instrument.

4. Close the cartridge cover and tray cover.

Note: The vial caps may contain impurities that can be detected with the Anion

Analysis Kit, therefore wash the caps before use.

Using the Anion Analysis Kit

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Conditioning a New Capillary

After installing a new capillary, rinse the capillary for one minute with Conditioner — Na. Wait four

minutes, then rinse for 30 seconds with Conditioner — Na. Rinse for one minute with Rinse

Solution. Use 20 psi of pressure for all rinses.

Storing the Capillary

After use, store the capillary on the instrument or in the original capillary storage box, with both

ends submerged in Rinse Solution. Do not allow the capillary ends to dry, because the capillary

may become plugged.

After a long storage period, or at the start of each day, condition the capillary using the Capillary

Conditioning method described in Running Methods on page 11.

Caution: Do not share capillaries between applications. If the capillary has been used for

cation analysis, do not use it for anion analysis.

Preparing the Buffer Trays

Use the correct vials and caps for your system:

• For the P/ACE MDQ system–use glass vials and red caps

• For the P/ACE MDQ plus system–use universal vials and blue caps

Replace all vials after twenty runs or after 24 hours inside the instrument. The increment option in

the method can be used to automatically increment the vials every twenty runs on both buffer

trays.

1. Fill the vials with equal volumes of each reagent in Tab le 1 -1 and position them in the

buffer trays (refer to Figure 1.1).

Use the correct volume for your system:

• For the P/ACE MDQ system–1.4 mL

• For the P/ACE MDQ plus system–1.5 mL

2. In the Waste position, place a vial partially filled with Rinse Solution.

Use the correct volume for your system:

• For the P/ACE MDQ system–700 µL

• For the P/ACE MDQ plus system–600 µL

3. Close each vial with a clean cap.

Using the Anion Analysis Kit

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4. Load the Inlet Buffer and Outlet Buffer trays in the instrument.

Note: The rinse solution used in this kit is ultra-purified water specifically for capillary

electrophoresis analysis of ions.

Preparing the Sample

Depending on the concentration of the analytes, the sample should be injected as is or diluted.

Dilution should be done so that the final concentration of the sample anions is between 5 ppm

and 200 ppm. Special care should be taken to verify the pH of the sample, which should be

above pH 5. A 50 mM NaOH solution can be used to dilute the sample and adjust its pH value.

The Anion Internal Standard (I.S.) consists of 30 mM sodium octanoate (sodium caprylate),

which is equivalent to 4296 ppm of octanoate ion. The I.S. can be used in the quantitation of the

sample anions. To use it, dilute the I. S. by a factor of 50 with the sample. For example, mix 4 µL

of I.S. with 200 µL of sample to yield 86 ppm of octanoate ion.

Figure 1.1 Buffer Tray Configuration for Anion Analysis

Note: When analyzing fluoride and/or phosphate ions, fill a vial with Anion

Acid Rinse and place it in position F1 of the Inlet Buffer Tray. The use of the

Anion Acid Rinse is required when running the Anion Inorganic Test Mix to

avoid peak tailing.

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PCR Vial Setup–P/ACE MDQ System

Fill a PCR vial with 200 µL of test or sample mix. Make sure there are no air bubbles at the

bottom of the PCR vial. Air bubbles can affect the sample injection. If bubbles exist, centrifuge

the vials for 2 minutes at 1000 x g and repeat if necessary. Place the PCR vial in a PCR holder

equipped with a vial spring (Figure 1.2). Seal the PCR vial with a clean gray cap and place it in

the inlet sample tray.

Figure 1.2 PCR Vial Setup–P/ACE MDQ System

Item Description

1 Vial cap (PN144656)

2 PCR vial (PN 144709)

3 PCR vial spring (PN 358821)

4 PCR vial holder (PN 144657)

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