Mettler Toledo PureSpeed C18 Operating instructions

Brand: Mettler Toledo

Model: PureSpeed C18

Type: Operating instructions

PureSpeed

™

C18

Desalting Tips

Operating Instructions

Introduction

Rainin PureSpeed C18 Desalting Tips are an innovative solution for micro-scale reverse phase

desalting. The tips are manufactured in two types to accommodate universal and LTS pipettes.

Each tip contains a small C18 lter at its distal end, with the purpose of desalting peptides

prior to proteomic applications. The tips may be used with the appropriate manual and elec-

tronic single and multichannel pipettes. The protocol contained in this manual describes the

use of the tips when desalting peptides.

Description

There are two different tip types for C18: 10 μL universal tips and 20 μL LTS tips. The 10 μL

traditional tips can be used with universal manual and electronic pipettes, whereas the 10 μL

LTS tips are used with Rainin manual and electronic pipettes with LTS. 20 μL Rainin manual

and electronic pipettes are recommended for the LTS desalting tip version.

Preparation

Prior to carrying out the desalting protocol, prepare the following solutions:

• Tip activation solvent – 100% Acetonitrile

• Tip washing solution – 0.1% (v/v) Trifluoroacetic Acid

• Tip elution solution – 0.1% (v/v)Trifluoroacetic Acid / 90% Acetonitrile

Sample Preparation – Prepare the sample in 0.1% (v/v) TFA. Optimal binding occurs within

this sample buffer.

Pipette Protocol (note the C18 protocol is included in the E4 XLS PureSpeed program)

1. Tip Loading –Mount a C18 tip to the pipette shaft (see table for recommended pipettes) by

pressing the shaft into the wide orifice on the tip. Make sure the tip is securely attached to

the pipette so that there is adequate sealing for up-and-down pipetting.

2. Tip Activation – Set the pipette volume setting to 10 μL or 20 μL depending on the pipette

nominal volume. Pipette 10 μL or 20 μL of tip activation solvent up-and-down five times,

dispensing all solvent from the tip. This action wets the tip and is required to bind the pep-

tide onto the C18 filter.

3. Tip Equilibration – Pipette 10 μL or 20 μL of tip washing solution up-and-down five times.

This step removes the majority of acetonitrile from the filter and prepares the resin for

sample binding. Dispense all solvent from the tip

4. Sample Binding – Pipette 10 μL or 20 μL of peptide sample up-and-down five times. This

step immobilizes the peptides to the C18 silica for washing in the next step.

5. Sample Washing – Pipette 10 μL or 20 μL of tip washing solution up-and-down five

times. Using fresh Tip Washing Solution each time, repeat this step two times. This step

removes the salt from the sample.

6. Sample Elution – Pipette 1 μL to 5 μL of tip elution solution up-and-down five times. This

removes the peptides from the C18 filter and elutes them into the tip elution solution.

7. Dispense sample into an appropriate container ready for the next steps in your workflow.

PureSpeed C18

Desalting Tips

PureSpeed C18

Troubleshooting

Problem Possible Cause Suggested Remedy

Tip does not aspirate

liquid

Tip is not mounted correctly, sealing is com-

promised

Make sure that the tip is securely fitted

onto the shaft or nozzle

Pipette seal has malfunctioned Replace seal

Electronic pipette is not charged Charge electronic pipette before use

Tip does not dis-

pense liquid

Tip is not mounted correctly, sealing is com-

promised

Make sure that the tip is securely fitted

onto the shaft or nozzle

Significant back pressure Use pipette blowout to dispense the

residual liquid

Tip does not bind

peptide

Sample contains organic solvents Evaporate solvents using lyophiliza-

tion or centrifugal evacuation

Peptides are not sufficiently hydrophobic to

bind tip

Change pH to modify peptide hydro-

phobicity

Tip has not been pre-wetted with acetonitrile,

or acetonitrile has evaporated from the tip

Make sure that the tip is activated with

100% acetonitrile before use

Tip does not elute

peptide

Peptides are too hydrophobic Use a elution solvent of higher elutro-

pic strength

Final sample is not

sufficiently desalted

Insufficient washing steps in C18 protocol Incorporate additional washing steps

prior to elution

Specifications

Pipette tip: Polypropylene

Media: C18: Irregular, 120 μm, 60 Å pore size

Max Tip Volume: 10 μL universal / 20 μL LTS

Absorptive bed: C18: 0.91 μL

Length: 1.05” universal / 1.65” LTS

Capacity: Up to 10 μg

Operating Temperature: 4°C – 70°C

Working pH Range: 2 to 13

Mass Spectrometry Data

Fig 1: Protein Sequence: Bovine Serum Albumin

220

For more information

Rainin Instrument, LLC.

7500 Edgewater Drive

Oakland, CA 94621

Sales +1 510 564 1600

Service +1 510 564 1600

Subject to technical changes

Printed in USA 9920414 Rev A

a METTLER TOLEDO Company

www.mt.com/purespeed

Rainin, Pipetting 360°, LTS, E4, XLS, and PureSpeed are trademarks of Rainin Instrument, LLC and

Mettler-Toledo, Inc.

Ordering Information

Catalog No.

MT Order

No.

Description

PureSpeed Tips and Accessories

PT-LC18-12 17014043 10 µL PureSpeed C18 Desalting Tips, LTS, 12 Tips

PT-C18-12 17014044 10 µL PureSpeed C18 Desalting Tips, Universal, 12 Tips

PT-LC18-96 17014045 10 µL PureSpeed C18 Desalting Tips, LTS, 96 Tips

PT-C18-96 17014046 10 µL PureSpeed C18 Desalting Tips, Universal, 96 Tips

PT-LC18-960 17014047 10 µL PureSpeed C18 Desalting Tips, LTS, 960 Tips

PT-C18-960 17014048 10 µL PureSpeed C18 Desalting Tips, Universal, 960 Tips

Recommended Rainin Pipettes

L-20XLS 17011785 Pipet-Lite XLS – LTS 2.0 to 20 µL

SL-10XLS 17011792 Pipet-Lite XLS – Universal 0.5 to 10 µL

L8-20XLS+ 17013803 Pipet-Lite XLS+ 8-channel – LTS 2.0 to 20 µL

L12-20XLS+ 17013808 Pipet-Lite XLS+ 12-channel – LTS 2.0 to 20 µL

E4-20XLS 17012306 E4 XLS – LTS 2.0 to 20 µL

SE4-10XLS 17012346 E4 XLS – Universal 0.5 to 10 µL

E8-20XLS+ 17013793 E4 XLS+ 8-channel – LTS 2.0 to 20 µL

E12-20XLS+ 17013798 E4 XLS+ 12-channel – LTS 2.0 to 20 µL

PureSpeed Accessories

PT-E4-SD 17012587 E4 XLS Micro SD Card with PureSpeed Protocol

Mettler Toledo PureSpeed C18 Operating instructions

Mettler Toledo PureSpeed C18 Operating instructions

Mettler Toledo PureSpeed C18 Operating instructions

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