[38] Improvement of efficiency of commercial spinning-cup sequenators

As, Bhown; Jc, Bennett

doi:10.1016/s0076-6879(83)91040-6

Cited by 2 publications

(1 citation statement)

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“…Currently most commercial protein sequencing systems employing the Edman degradation method use high-performance liquid chromatography (HPLC) with ultra-violet (UV) absorbance detection for the analysis of the phenylthiohydantoin (PTH) amino acids products that are produced from this process. Although systematic improvements over the years have been made to this novel method of protein sequencing [3,4,5,6,7], the choice of detection still suffers markedly from lack of speed, sensitivity, and structural information of nonstandard amino acids. Due to these limitations in detection there have been a number of alternative methodologies -mainly involving mass spectrometry (MS) -developed for the analysis of PTH amino acid derivatives.…”

Section: Introductionmentioning

confidence: 99%

Rapid separation of phenylthiohydantoin amino acids: ambient pressure ion-mobility mass spectrometry (IMMS)

2003

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An electrospray ionization (ESI) ambient pressure ion-mobility spectrometer (APIMS) interfaced to an orthogonal reflector time-of-flight mass spectrometer (TOFMS) was evaluated for the first time as a detector for the identification of phenylthiohydantoin (PTH)-derivatized amino acids, the final products in the Edman sequencing process of peptides and proteins. The drift and flight times of the twenty common PTH amino acids were characterized by a well-defined 2-D mobility/mass spectral pattern. The combination of mobility/mass modes of analysis gave rise to a unique trend-line formation for the series of PTH amino acids. In addition, each PTH amino acid had a unique reduced mobility constant K(o), thus enabling the differentiation of all the amino acid derivatives including the PTH-leucine and PTH-isoleucine isomers. More importantly it was shown that it was possible to resolve a complete reference mixture of PTH amino acids in a single experimental run in less than 1 min. Detection limits for the PTH amino acids were found to range from 1.04 to 3.52 ng; indicating that the limits of detection were less than 17.0 pmol for all of the PTH amino acids.

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Section: Introductionmentioning

confidence: 99%