Fmoc, 9-fluorenylmethoxycarbonyl monitoring

Figure 9.140 HPLC elution profile of a sample taken from a discontinuous assay of crude mouse spleen extract, using iV,-fluorenylmethoxycarbonyl-EEY(P)AA [Fmoc-EEY(P)AA] as substrate. Chromatography was performed on a C g Novapak column (10 cm X 8 mm) eluted isocratically with 36% acetonitrile-water (0.1% TFA) (v/v) at a flow rate of 2 mL/min. Peaks are due to (A) methanol and methanol-soluble compounds derived from the sample of crude homogenate, (B) Fmoc-EEY(P)AA (1238 pmol), and (C) Fmoc-EEYAA (195 pmol). Appropriate controls showed that no interfering compounds eluted in the position of the peptides. Inset Fluorescence monitoring of HPLC of Fmoc-EEYAA (75 fmol) eluted isocratically with 36% acetonitrile-water (0.1% TFA). Excitation and emission wavelengths, 268 and 307 nm, respectively, with gain X 100 and 10 mV chart scale. (From Nash et al., 1993.)...

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