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)