A fluorimetric method for the determination of tryptophan in animal tissues.
Tryptophan, tryptamine and peptides containing N-terminal tryptophan give two highly fluorescent products on treatment with dithiothreitol and acid ninhydrin reagent 1 or 2. The first fluorescent product (product A) gives an emission at 500nm on activation at 390-400nm and is stable for 20min. The second product (product B), which gives an emission at 530nm on activation at 470nm, is detectable within 1h after the reaction. It gives almost maximum intensity in 4h and is stable for at least 48h. Except lysine, which in equimolar amounts gives less than 1% of a product similar to product B, no other naturally occurring amino compounds give fluorescent products. A procedure is given for the determination of 0.05-34nmol of tryptophan in tissue extracts. By using this procedure rat brain was found to contain 17.56+/-0.76 (s.e.m.) nmol/g wet wt.
D000085:Acetates; D000818:Animals; D001923:Brain Chemistry; D001933:Brain Stem / Q000032:analysis; D002021:Buffers; D002951:Citrates; D004027:Diencephalon / Q000032:analysis; D004229:Dithiothreitol; D007031:Hypothalamus / Q000032:analysis; D007202:Indicators and Reagents; D007475:Ion Exchange Resins; D008239:Lysine / Q000032:analysis; D008722:Methods; D051379:Mice; D010455:Peptides / Q000032:analysis; D051381:Rats; D013050:Spectrometry, Fluorescence; D014363:Tryptamines / Q000032:analysis; D014364:Tryptophan / Q000032:analysis* / Q000302:isolation & purification
D000085:Acetates; D002021:Buffers; D002951:Citrates; D007202:Indicators and Reagents; D007475:Ion Exchange Resins; D010455:Peptides; D014363:Tryptamines; D014364:Tryptophan; D008239:Lysine; D004229:Dithiothreitol