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.
- 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
- 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
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 (Edman's reagent).
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