Method for separating diphenylurea (DPU) and phenylthiohydantoin

Details Method for separating diphenylurea (DPU) and phenylthiohydantoin Method for separating diphenylurea (DPU) and phenylthiohydantoin

1990-03-13

1992-09-01

Housel, James C.

Chemistry: analytical and immunological testing

Peptide, protein or amino acid

Amino acid or sequencing procedure

436 86, 436 87, 210656, 2101982, G01N 3300, B01D 1508

Patent

active

051438518

ABSTRACT:
A reverse-phase HPLC method is disclosed which allows the separation of DPU and PTH-Trp and, therefore, the correct assignment of tryptophan residues in an amino acid sequence. The method is based on a modification of the conventional HPLC gradient commonly used to elute and separate all PTH amino acids of interest in automated and manual sequencing. In one embodiment, using automated instruments manufactured by Applied Biosystems, gradient modification is achieved by changing the manufacturer-supplied gradient program which controls the operation of the HPLC pumps in the instrument. Using the methods of the present invention, the correct and reliable assignment of tryptophan residues is possible and was reproducible over time, even with small sample sizes.

REFERENCES:
patent: 4665037 (1987-05-01), Stolowitz

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