Flow Rate 0.4 mL/min	A	B
Time	Methanol: Formic acid (10%: 0.1%)	Methanol: Formic acid (50%: 0.1%)
0–6.5	10–30	90–70
6.5–7	30–100	70–0
8–8.5	100–10	0–90
8.5–12.5	10	90

Fluorescence is detected using a detector by excitation at 335 nm and emission at 440 nm. Retention was used to distinguish amino acids [32].