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Phenylisothiocyanate (PITC)


High-purity reagent for pre-column quantitation of amino acids by reverse-phase HPLC.

Pierce PITC (phenylisothiocyanate)

Thermo Scientific Pierce Phenylisothiocyanate (PITC) is a high-purity reagent for pre-column quantitative derivatization of amino acids by reverse-phase HPLC.

Phenylisothiocyanate (PITC), also known as Edman’s Reagent, enables the sequential degradation of amino acids in a polypeptide chain, yielding primary structural information. PITC reacts readily with amino acids at alkaline pH. Precolumn derivatization results in phenylthiocarbamyl derivatives (PTC-amino acids) that can be separated and quantified using reverse-phase HPLC. This method produces stable products with all amino acids, including proline. PITC is volatile, making it possible to remove excess reagent in vacuo, thereby minimizing the possibility of reagent interference. Detection of picomole quantities of the derivatives can be achieved using a UV detector at 254nm. PITC derivatization followed by reverse-phase chromatography can be used for identification and quantitation of methylated, halogenated, phosphorylated and sulfonated amino acids.

Highlights:

  • For pre-column derivatization of amino acids for picomole detection levels in HPLC
  • Reacts with amine groups of amino acids in 5 to 10 minutes at room temperature to yield products that are detectable by UV absorbance (λmax = 254nm)
  • Produces phenylthiocarbamyl (PTC) derivatives that can be separated and quantified in 30 minutes using reverse-phase HPLC
  • Produces stable products with all amino acids, including proline

Reagent Properties:

  • Alternative names: PITC, Phenyl Isothiocyanate, Edman's Reagent
  • Chemical name: Isothiocyanatobenzene
  • Reactive toward: Primary amines (–NH2)
  • Chemical formula: C7H5NS
  • CAS number: 103-72-0
  • Molecular weight: 135.19
  • Form: Clear, colorless to pale yellow liquid

Product Details:

Unlike Fmoc-chloride, PITC does not yield disubstituted tyrosine or histidine derivatives. PTC-amino acids demonstrate improved stability at pH 5-7.5 as well as increased stability at room temperature over o-phthalaldehyde (OPA)-amino acid adducts. Also, unlike OPA, PITC enables the direct analysis of secondary amino acids.

Chemical structure of PITC, phenylisothiocyanate, Edman's Reagent Chemical structure of PITC (Edman's reagent).

References:

  1. Edman, P. (1950). Preparation of phenylthiohydrantoins from natural amino acids. Acta Chem Scand 4:277-82.
  2. Edman, P. and Begg, G. (1967). A protein seqenator. Eur J Biochem 1(1):80-91.
  3. Heinrikson, R.L. and Meridith, S.C. (1984). Amino acid analysis by reverse-phase high-performance liquid chromatography: Precolumn derivatization with phenylisothiocyanate. Anal Biochem 136:65-74.
  4. Schoze, H. (1985). Determination of phenylthiocarbamyl amino acids by reverse-phase high-performance liquid chromatography. J Chromatogr 350:453-60.
  5. Ebert, R.F. (1986). Amino acid analysis by HPLC: Optimized conditions for chromatography of phenylthiocarbamyl derivatives. Anal Biochem 154:431-5.
  6. Cohen, S.A. and Strydom, D.J. (1988). Amino acid analysis utilizing phenyliosthiocyanate derivatives. Anal Biochem 174:1-16.

Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
26922 PITC (Edman's Reagent)
Formulation: Phenylisothiocyanate (at least 99.5% pure)
Sufficient For: Derivatizing hundreds of amino acid samples for typical HPLC analysis
10 x 1mL Product Instructions for product #26922 PITC (Edman's Reagent) MSDS for product #26922 PITC (Edman's Reagent) Certificate of Analysis for product #26922 PITC (Edman's Reagent) $108.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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