Team:UFlorida/Experiments

HPLC Conditions

Eluent A

2.5 : 97.5 % (v/v) acetic acid: H2O, pH 3.8

*pH was adjusted by addition of 1 mol L-1 KOH

Eluent B

80 : 20 % (v/v) acetonitrile: H2O

Gradient Elution (A>B)

80:20

50:50

0:100

80:20

Time Scale(min) @ FR (1ml/min)

0-25

25-31

31-33

33-36

Analysis

Initial Standard Run of Tryptophan

MW of compounds:
Tryptophan MM: 204.225 g/mol
Indole-3-pyruvic acid MM: 203.197 g/mol
Indole-3-acetaldehyde MM: 159.188 g/mol
Tryptophol MM: 161.20 g/mol

Tryptophan and Indole-3-pyruvic acid

Tryptophan and indole-3-pyruvic acid are too close in MM that unintended reactions during the run would make it difficult to differentiate the two.

References

King ED, Ward MK, Raney DE, 1954. Two simple media for the demonstration of pyocyanin and fluorescin. Journal of Laboratory and Clinical Medicine 44, 301–7.