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
1) Add 20 μL of CH3
CN to 20 μL of serum
2) Stir (2 to 3 sec)
3) Centrifuge (10,000G , 2min)
4) Skim supermatant and inject
Sample : Indomethacin
Column : Shodex Asahipak ODP-50 6E (6.0mmID*150mm)
Eluent : 25mM Sodium phosphate buffer(pH10.0)/CH3CN=65/35
Flow rate : 1.0mL/min
Detector : Fluorescence detecotr(Ex.298nm , Em.375nm)
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