Thermo Scientific
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43
Thermo Scientific
Peptide Retention Standards
Peptide Retention Standard
for Reverse-Phase HPLC
Increases the efficiency of peptide elution profile
predictions.
A simple, quantitative method for predicting peptide
retention times was developed by Guo,
et. al.
1-3
Retention
times are predicted by totaling the values that represent the
contribution in minutes of each amino acid residue and the
peptide terminal groups.
Retention time is dependent upon the molecular weight of
the peptide. The effect on retention is relatively unimportant
with a small peptide, but it increases with the size of the
molecule. The accuracy of predicting peptide retention time
significantly decreases beyond 20 residues.
To ensure accuracy, a peptide standard is used to correct
for instrument variation, column aging, n-alkyl chain length
variation and ligand density.
By using a Thermo Scientific Peptide Retention Standard,
you can:
• Determine the relative order of peptide elution of a
complex mixture
• Increase the efficiency of predicting peptide elution
profiles
• Save time in peptide purification
• Simplify identification of specific peptides in a complex
mixture
• Predict the HPLC retention time for peptides of known
amino acid composition on reverse-phase HPLC columns
• Monitor column performance – efficiency, selectivity and
resolution during column aging
• Compare reverse-phase columns from different
manufacturers
• Evaluate reverse-phase supports of varying n-alkyl chain
lengths and ligand densities
References
1. Guo, D.,
et al.
(1985).
Proceedings of the Ninth American Peptide Symposium,
Published by Pierce, Rockford, Illinois, page 23.
2. Guo, D.,
et al.
(1986).
J. Chromatogr.
359,
499-517.
3. Guo, D.,
et al.
(1986).
J. Chromatogr.
359,
519-532.
4. Mant, C.T. and Hodges, R.S. (1986).
L.C. Magazine Liq. Chrom. and HPLC
4(3),
250.
5. Guo, D.,
et al.
(1987).
J. Chromatogr.
386,
205-222.
Ordering Information
Product # Description
Pkg. Size
TS-31700
Peptide Retention Standard, S1-S5
1 vial
Contains: 5 C-terminal amide decapeptides,
4 of which are N-acetylated with the
sequence variation as follows:
AC-Arg-Gly-X-X-Gly-Leu-GIy-Leu-GIy-Lys-
Amide; GIy
3
-Gly
4
-AIa
3
-GIy
4
, VaI
3
-GIy
4
and Val
3
-
Val
4
. The fifth peptide, AIa
3
-GIy
4
, contains a free
N-amino group. This mixture will provide
100-200 injections at 0.1 AUFS at 210 nm.
Peptide Retention Time Calibration
Mixture
Heavy peptide mixture for column assessment and
prediction of peptide retention times.
• Assessment of chromatography and MS instrument
performance
• Prediction of peptide retention across multiple
instrument platforms
• Prediction of peptide retention time from sequence using
calculated hydrophobicity factor
• Optimization of scheduled MS acquisition windows for
improving quantification and increased multiplexing
• Internal standard to normalise for variation in retention
times and peak intensities between runs
Ordering Information
Product # Description
Pkg. Size
TS-88320
Peptide Retention Time Calibration
50µL x1
Mixture, 0.5pmol/µL
TS-88321
Peptide Retention Time Calibration
200µL x1
Mixture, 5pmol/µL
