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For more information, or to download product instructions,
Thermo Scientific HPLC Ion Pair Reagents
Thermo Scientific Hypercarb Columns
100% porous graphitic carbon for extended
separation capabilities
• Exceptional retention of very polar analytes
• Separates structurally related substances
• pH stable from 0 to 14
• Ideal for high temperature applications
Trifluoroacetic Acid (TFA)
1 ml ampules allow you to make a fresh 0.1% TFA solution in
seconds!
Thermo Scientific Trifluoroacetic Acid (TFA) is the most
commonly used ion pairing agent in reverse-phase peptide
separations because TFA:
• Sharpens peaks and improves resolution
• Is volatile and easily removed
• Has low absorption within detection wavelengths
• Has a proven history
Highlights:
Purity
• > 99.5% TFA purity and exceptional clarity, allowing
sensitive, nondestructive peptide detection at low UV
wavelengths in reverse-phase HPLC protein and peptide
separation systems
High-performance packaging
• Our TFA is packaged under nitrogen in amber glass
ampules or bottles with protective Teflon – TFE-lined
fluorocarbon caps to ensure TFA integrity
Economical convenience
• Choose the TFA format that works best for your
application. In just a few seconds, 1 ml ampules can be
used to prepare 1 liter of fresh 0.1% v/v trifluoroacetic acid
solution for the mobile phase in reverse-phase
chromatography
Applications:
• Ion pair reagent for reverse-phase HPLC
1-3
• Protein/peptide sequencing
4-7
• Protein/peptide solubilizing agent
4-7
• Solid-phase peptide synthesis
8
• Amino acid analysis
Making 0.1% Solutions of Trifluoroacetic Acid
For complex peptide separations, the key to success can be
to vary selectivity. Varying mobile phase composition on the
same column can change selectivity enough to resolve
peptides that would otherwise overlap. Trifluoroacetic acid is
the most frequently used modifier for peptide separations in
reverse-phase HPLC. The TFA concentration usually
specified is 0.1%. For reproducible separations from
run-to-run or from lab-to-lab, it is essential to make TFA
concentrations the same.
Trifluoroacetic acid concentration can and should be
specified as either “w/v” (weight/volume), or as “v/v”
(volume/volume). The w/v specification designates that the
TFA is to be weighed and added to a volume of mobile phase
(e.g. 0.1% TFA w/v requires one gram of TFA per liter). The v/v
specification designates that the TFA is to be measured by
volume (e.g. 0.1% TFA v/v requires one ml of TFA per liter).
Because the density of trifluoroacetic acid is 1.53 g/ml the
difference between 0.1% TFA (w/v) and 0.1% TFA (v/v) is more
than 50%. For the sake of reproducibility, it is essential for
authors of a method to specify, and for users of a method to
know, whether the TFA concentration is given as “w/v”
or “v/v”.
References
1. Chicz, R.M. and Regnier, F.E. (1990) Methods Enzymol.
182,
392-421.
2. Zarember, K.A., et al. (2002) Infect. Immun.
70,
569-576.
3. Lassy, R.A. and Miller, C.G. (2000) J. Bacteriol.
182,
2536-2543.
4. Smith, B.J. (1997) Protein Sequencing Protocols. Humana Press.
5. Allen, G. (1989) Sequencing of Proteins and Peptides, Second Revised Addition.
Elsevier.
6. Backstrom, J.R., et al. (1996) J. Neurosci.
16,
7910-7919.
7. Hermann, P.M., et al (2000) J. Neurosci.
20,
6355-6364.
8. Stuart, J.M. and Young, J.D. (1984) Solid Phase Peptide Synthesis, Second
Edition. Pierce Chemical Company.
Ordering Information
Product # Description
Pkg. Size
✖
TS-28901
Trifluoroacetic Acid, Sequencing Grade
500 ml
✖
TS-28902
Trifluoroacetic Acid, Sequencing Grade
10 x 1 g
✖
TS-28903
Trifluoroacetic Acid, Sequencing Grade
100 g
✖
TS-28904
Trifluoroacetic Acid, Sequencing Grade
10 x 1 ml
ampules
✖
Additional hazardous handling charge.
A planar compound can align itself
closely with the Hypercarb
®
surface resulting
in more interaction and retention.
Electrostatic charge of polar analyte
approaching the graphite surface
Electrons in the polarizable surface of the
graphite attracted towards positive charge
Electrons in the polarizable surface of the
graphite repelled away from a negative charge
Electrostatic charge of polar analyte
approaching the graphite surface
A non-planar molecule cannot align itself
as closely with the Hypercarb surface resulting
in less interaction and retention.
Charge-induced dipole at
the graphite surface
