Indomethacin was analyzed using Asahipak ODP-50 6E (a column for reversed phase chromatography). Indomethacin is a steroid-free, anti-inflammation drug used for rheumatism, inflammation of arthritis and podagra and is known to have strong side effects on the central nervous system and digestive organs. This is an example of a high sensitive analysis of indomethacin by using the principle that it converts to a fluorescent substance by alkaline hydrolysis at high temperature.

1) Use an ODP column with an eluent of pH10.
2) Set a heating coil at postcolumn.
3) Only one pump is necessary. Since the eluent is alkaline, postcolumn mixing is unnecessary.
4) The indomethacin is converted into fluorescent substance by alkaline hydrolysis in the heating coil, a highly selective and sensitive analysis is possible.

(Sample pre-treatment)
1) Add 20 μL of CH ₃ CN to 20 μL of serum
2) Stir (2 to 3 sec)
3) Centrifuge (10,000 G , 2 min)
4) Skim supermatant and inject


Sample : Indomethacin in Serum

Column       : Shodex Asahipak ODP-50 6E (6.0 mm I.D. x 150 mm)
Eluent       : 25 mM Sodium phosphate buffer (pH10.0)/CH3CN=65/35
Flow rate    : 1.0 mL/min
Detector     : Fluorescence (Ex. 298 nm , Em. 375 nm)
Column temp. : Room temp.
Post column  : Heating coil at 140 °C is followed with cooling coil reaction at 15 °C

H. Kubo, Y.Umiguchi, T.Kinoshita, Chromatographia Vol.33, No.7/8, 1992