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. 2014 Mar;55(3):573–582. doi: 10.1194/jlr.D044230

Fig. 2.

Fig. 2.

Optimization of aldehyde derivatization. Concentration of the different assay components, pH, and heating parameters were varied to optimize the derivatization of C13-al (triangles) and C17-al (open squares) (1.5 and 2 nmol, respectively). Fluorescent derivatives were directly separated by RP-HPLC, without dilution with methanol, and the integrated area of the signal was plotted against the variable parameter, while other parameters (1.3 M ammonium acetate, 35.7 mM CHD, 13.3% (v/v) acetic acid, 80°C temperature, 90 min heating time, 40% (v/v) methanol, based on initial tests) were kept constant, unless otherwise mentioned. A: Effect of ammonium acetate. B: Effect of CHD. C: Effect of acetic acid. D: Effect of temperature (1.6 M ammonium acetate, 14% (v/v) acetic acid, 60 min heating time). E: Effect of heating time (1.6 M ammonium acetate, 14% (v/v) acetic acid). F: Effect of methanol (1.2 M ammonium acetate, 14% (v/v) acetic acid, 65°C temperature, 60 min heating time).