UHPLC Analysis of Aflatoxins on Titan C18 with TFA Derivatization and Fluoresence Detection

sample preparation

200 μL of standard solution of aflatoxins was mixed with 300 μL of derivatization solution (7:1:2 water:acetic acid:trifluoroacetic acid). (The mixture was kept at 65 °C for 25 minutes, then cooled down to room temperature. 550 μL of water was added.)

column

Titan C18, 10 cm x 2.1 mm I.D., 1.9 μm particles (577124-U)

mobile phase

[A] 5% acetonitrile in water; [B] acetonitrile.

gradient

10 to 30% B in 3 min; to 100% B in 2 min; held at 100% B for 2 min; to 10% B in 0.5 min; held at 10% B for 2.5 min

flow rate

0.4 mL/min

column temp.

35 °C

detector

Fluorescence, 360/440 Ex/Em

injection

20 μL

sample

standard solution of aflatoxins contained 1.2 ng/mL each of afltoxin B1 and aflatoxin G1, and 0.3 ng/mL each of aflatoxin B2 and afltoxin G2 in acetonitrile:water (84:16)