A rapid and sensitive multisample identification method for methylthiohydantoin amino acids (MTH): Improved thin-layer chromatographic separation on a microscale
✍ Scribed by Klaus D. Kulbe; Yolanda Maria Nogueira-Hattesohl
- Publisher
- Elsevier Science
- Year
- 1976
- Tongue
- English
- Weight
- 780 KB
- Volume
- 72
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Two solvent systems have been established which are well suited for twodimensional separation of 23 methylthiohydantoin-amino acids obtained in Edman degradation with methyhsothiocyanate. Using a fluorescent indicator and doublefaced polyamide layers, it is also possible to separate and to identify at least 10 individual MTHI samples on a single 5 x 5 cm sheet at subnanomolar level (0.05-0.2 nmol) within 30 min by consecutively applying both solvents in identical direction. The methyhhiohydantoins of chemically modified cysteine and methionine derivatives most commomy obtained in primary structure determination are included into the standard mixtures. In case of insufficient separation of the Leu, lie, Phe, Val, and Thr derivatives, a third solvent was used. Synthesis and purification of MTH-AE'-Cys, MTH-His, MTH-MetSO% and MTH-Ser are described. The new method is a valuable adjunct in manual and automated sequence determination.