Amino Acids, Peptides and Proteins: An Introduction by H.D. Jakubke, J. Jeschkeit

By H.D. Jakubke, J. Jeschkeit

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In the gas chromatographic analysis of Dnp-amino acid methyl esters, amounts as small as 10-14 mol of amino acid may be detected. This extreme sensitivity is achieved with an electron capture detector, making use of the high electron affinity of the Dnp group. The separation of diastereoisomeric amino acid derivatives by gas chromatography has already been described (on p. 42). 48 Amino Acids Table 10. c: Cl.. ~a! 0 0 ~'~ '- ~ ..... ::1 \0 l!. ~ ~ '- <-J ~ E C) g I 0 Time 15 20 25 Jamin Figure 12.

This was a considerable achievement. The accuracy ( = 3 per cent) was the highest achieved up to that t-ime and very little material was used. However, the analysis of each sample took a week. MooRE and STEIN were able to perform the analysis in a shorter time by changing to ion exchange chromatography. The most suitable ion-exchangers for amino acid analysis are the Dowex resins (Dowex 50 X 8 and Dowex 50 X 4) which are crosslinked with 8 and 4 per cent divinylbenzene, respectively, and sulphonated polystyrene resins of small bead size (Amberlite resins), which \Yere used in the first automatic amino acid analysers 56 • Figure 10 illustrates the principle of the first automatic analysers developed by SPACKMANN, MooRE and STEIN.

Ultra-violet region above wavelengths of 220 nm, the aromatic amino acids tryptophan, tyrosine, phenylalanine and histidine show characteristic maxima above 250 nm. The high molar extinctions of tyrosine and tryptophan facilitate rapid quantitative determination of these amino acids in proteins. In the IR spectra of amino acids the normal NH-stretching frequencies in the region of 3300-3500 cm-1 are absent. Instead, absorption at 3070 cm- 1 6-1~ E. I ~1 3 \ ~ \ 'I\ 0zz5 245 265 285 nm315 a 2 1 I I \I I I \ ', \ ~ t( I v / ; ' \ b \ 0225 2lr5 265 285 nm 315 Figure 5.

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