HPLC Ion Pair Reagents

Pierce™ Heptafluorobutyric Acid (HFBA), Sequencing Grade (Thermo Scientific™)

Thermo Scientific Pierce Heptafluorobutyric Acid (HFBA) is manufactured and tested to meet strict specifications that ensure superior performance for use as an ion-pairing agent.

Features of HFBA:

Greater than 99.5% purity—allows sensitive, nondestructive peptide detection at low UV wavelengths in peptide separation systems
High-performance packaging—HFBA packaged under nitrogen
Quality control—tested for overall purity, chain-length purity, water content, sulfate content and absorptivity at three different UV-wavelengths

Applications:
• Protein/peptide sequencing
• Protein/peptide solubilizing agent
• Solid-phase peptide synthesis
• Amino acid analysis

Pierce HFBA is manufactured to the highest specifications to ensure the integrity of your data, maximize sensitivity in your assay and to prolong the life of your equipment.

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Pierce™ Heptafluorobutyric Acid (HFBA), HPLC Grade

Pierce™ Formic Acid, LC-MS Grade (Thermo Scientific™)

Thermo Scientific Pierce Formic Acid is high-purity solvent supplied in bottles or ampules as a convenient, contamination-free alternative for preparing elution solvents for HPLC separations of protein and peptides. This product is packaged in amber glass, pre-scored, nitrogen-flushed ampules to protect formic acid from light, moisture and contamination.

Features of Formic Acid:

Greater than 99% pure formic acid—consistent LC baselines, no potential interference introduced in LC or MS applications, and no signal suppression in the mass spectrometer
Convenient format—ampule packaging simplifies the preparation of gradient and isocratic mobile phases containing 0.1% (v/v) formic acid in water or acetonitrile; the contents of a single vial in a final volume of 1 L of solvent yields a mobile phase of the most common formic acid concentration.

General properties of formic acid
• Molecular formula: HCOOH (CH2O2)
• Molecular weight: 46.02
• Density: 1.22 g/mL
• CAS number: 64-18-6
• Refractive index: 1.3701–1.3721(20°C)
• Flash point: 69°C
• Freezing point: 8°C

Specifications of Thermo Scientific Pierce Formic Acid
• Visual: Clear liquid, free of particulate matter
• Identity (IR): Must show only peaks characteristic for the compound
• Purity: > 99%
• Refractive index: 1.3701 to 1.3721 (20°C, 589 nm)

Pierce Formic Acid is sealed in amber glass ampules under a dry nitrogen atmosphere. A pre-measured aliquot of acid greatly simplifies preparation of liter quantities of mobile phases at the standard 0.1% formic acid concentration. The quality of this formic acid coupled with either glass-ampule bottle packaging provides reliability and convenience that adds value to both the chromatographic and MS results.

Pierce Formic Acid is specially purified and tested to the highest specifications to ensure the integrity of your data, maximize sensitivity in your assay and to prolong the life of your equipment.

Formic acid is a common component of reverse-phase mobile phases that provide protons for LC/MS analysis. The presence of a low concentration of formic acid in the mobile phase is also known to improve the peak shapes of the resulting separation. Unlike trifluoroacetic acid (TFA), formic acid is not an ion-pairing agent and it does not suppress MS ionization of polypeptides when used as a mobile-phase component.

For added safety when opening glass ampules, consider using our Ampule Breakers, which are disposable safety devices that protect the fingers when breaking open a glass ampule.

Pierce™ Trifluoroacetic Acid (TFA), Sequencing grade (Thermo Scientific™)

Thermo Scientific Pierce Trifluoroacetic Acid (TFA), Sequencing grade is manufactured and tested to meet strict specifications that ensure superior performance for use as an ion-pairing agent in reverse-phase peptide separations.

Features of Trifluoroacetic Acid (TFA), Sequencing grade:

High purity and exceptional clarity—allows sensitive, nondestructive peptide detection at low UV wavelengths in reverse-phase HPLC protein and peptide separation systems
High-performance packaging—TFA packaged under nitrogen in amber glass ampules or bottles with protective PTFE-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 of Trifluoroacetic Acid (TFA), Sequencing grade:
• Ion pair reagent for reverse-phase HPLC
• Protein/peptide sequencing
• Protein/peptide solubilizing agent
• Solid-phase peptide synthesis
• Amino acid analysis
• Making 0.1% solutions of trifluoroacetic acid (w/v vs. v/v)

Trifluoroacetic acid (TFA) is the most commonly used ion pairing agent for use in reverse-phase HPLC peptide separations because it sharpens peaks and improves resolution, is volatile and easily removed, has low absorption within detection wavelengths. It is manufactured to the highest specifications to ensure the integrity of your data, maximize sensitivity in your assay and to prolong the life of your equipment.

Making 0.1% Trifluoracetic Acid Solutions:
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".

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Pierce™ Trifluoroacetic Acid (TFA), LC-MS Grade

Pierce™ Triethylamine (TEA), Sequencing Grade (Thermo Scientific™)

Thermo Scientific Pierce Triethylamine (TEA) is a high-purity solvent that is used in certain HPLC protocols, such as weak anion exchange, to resolve certain tryptic peptides on a reverse-phase column.

Features of Thermo Scientific Pierce Triethylamine (TEA):

• Greater than 99.5% triethylamine purity, allowing sensitive peptide detection at low UV wavelengths in reverse-phase HPLC peptide separation systems
• Packaged in amber glass bottles with protective PTFE-lined fluorocarbon caps for reagent integrity

Triethylamine is an ion-pairing reagent that alters selectivity in reverse-phase HPLC separations. By pairing with peptides, it effectively sharpens peaks, resulting in improved peak resolution. Pierce Triethylamine has low UV absorbance properties to provide sensitive detection across all wavelengths when used as an ion-pair reagent in HPLC separations. The reagent is designed and tested to meet the special requirements for peptide sequencing and analysis.

Pierce Triethylamine is specially purified and each lot is tested to the highest specifications to ensure the integrity of your data, maximize sensitivity in your assay and to prolong the life of your equipment.