Waters MassPREP Peptide Mix Standard User Manual

[ CARE AND USE MANUAL ]

MassPREP Peptide Mixture

CONTENTS

I. INTRODUCTION

II. STORAGE AND STABILIT Y

III. RECONSTITUTION OF MASSPREP
PEPTIDE MIXTURE

INTRODUCTION

The peptide mixture contains nine peptides with a broad range of

polarities, and isoelectric points (pI). In addition, there is a void

volume marker (Table 1). All peaks are resolved under standard

reversed-phase LC conditions typically used for peptide separation

(Figure 1). Therefore the mixture can be used for the evaluation of

columns and LC systems dedicated for peptide separations.

IV. MSDS INFORMATION

V. ORDERING INFORMATION

Peak #

Component

Name

1 Allantoin (V0 marker) 0.76 158.0 4 4 0 - -

2 R A S G-1 4.78 1000.4938 9.34 RGDS PASSK P

3 Angiotensin frag.1-7 9.59 898.4661 7.35 DRVYIHP

4 Bradykinin 12.0 4 1059.5 613 12.00 RPPGFSPFR

5 Angiotensin II. 13.06 1045.5345 7. 35 DRVYIHPF

6 Angiotensin I. 14.92 1295.6 775 7.51 DRVYIHPFHL

7 Renin substrate 16.82 1757.9253 7.61 DRVYIHPFHLLVYS

8 Enolase T35 19.18 1871.9604 7.3 4 WLTGPQLADLYHSLMK

9 Enolase T37 21.55 2827.2806 3.97 YPIVSIEDPFAEDDWEAWSHFFK

10 Melittin 24.93 2845.7381 12.0 6 GIGAVLKVLTTGLPALISWIKRKRQQ

Retention Time

*(minutes)

Molecular Weight

**(g/mol)

pKa Peptide Sequence

Table 1

* BioSuite™ C18 3.5 µm PA-B Column (4.6 x 50 mm), mobile phase A: 0.1% TFA

in water, B: 0.08% TFA in acetonitrile, gradient: 0-50% B in 30 minutes, 0.96
ml/min, 40° C 214 nm. Retention times will change if different LC conditions,
column, and LC system are used.

** Monoisotopic molecular weight, MassLynx™ v. 4.0

3.5 µm PA-B

18

0.5

Enolase T37

Enolase T35

Angiotensin I

Angiotensin II

Allantoin

AU

RASG-1

0.0

0 10 20 30

For chromatographic conditions see Table 1.

Figure 1: Standard Peptide Mixture on BioSuite C18 3.5 µm PA-B Column
(4.6 x 50 mm).

Angiotensin frag. 1-7

Bradykinin

Renin substrate

Melittin

Minutes

[ CARE AND USE MANUAL ]

II. STORAGE AND STABILITY

The lyophilized powder is stable at room temperature until

the expiration date printed on the label. The sample can be

reconstituted in 0.1% aqueous formic acid (for electrospray or

LC/MS applications) or in 0.1% aqueous trifluoroacetic acid

(for MALDI and LC/UV applications), the solution is stable

for one week when stored at 8 °C. Stability can be extended

by freezing samples at -20 °C. Note: The contamination of

sample with endo- and exopeptidase enzymes may cause

rapid sample degradation.

III. RECONSTITUTION OF MASSPREP
PEPTIDE MIXTURE

The peptide mixture contains approximately 1.5 µg (~1 nmole)

of each peptide. When reconstituted in 100 µL of solvent (0.1%

aqueous trifluoroacetic acid, or 0.1% aqueous formic acid),

the final sample concentration is ~15 µg/ml per peptide.

The suggested sample injection volumes for LC columns with

different internal diameters are summarized in Table 2.

IV. MSDS INFORMATION

Waters Peptide Mixture contains various non hazardous peptides

in powder form. T hey are for laboratory use only; solely for

purposes of scientific experimentation, analysis, research

or development. Waters Peptide Mixture should be used only

as directed and in accordance with good laboratory practice.

All Waters Peptide Mixtures are non hazardous as defined by

OSHA’s Hazard Communication Standard, 29 CFR 1910.1200(c)

and Appendix A and B, and are thus exempt from the requirements

of the Hazard Communication Standard, including the requirement

to supply MSDSs.

V. ORDERING INFORMATION

Item Part Number Quantity

Peptide Mixture 186 0 023 37 1

Peptide Mixture (5-pack) 186002338 5

Reconstitution

Volume (µl)

Column I. D.

(mm)

Inje cti o n

Volume

(µl)

Inje cted M ass

Per Peptide (ng)

100 4.6 75.0 112 5 . 0 750.0

100 2 .1 15.6 234.5 156.3

100 1.0 3.5 53.2 35.4

100 0.32 0.4 5.4 3.6

100 0.18 0.1 1.7 1.1

100 0.075 0.02 0.3 0.2

Table 2: Recommended Mass Load for LC Columns Evaluation.

Avg. I njecte d

Mas s Per

Peptide

(pmol e) *

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©2014 Waters Corporation.Produced in the U.S.A. August 2014 715001703EN TC-PDF

MassPREP, MassLynx, and BioSuite

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