Detection of indomethacin by high-performance liquid chromatography with in situ electrogenerated Mn(III) chemiluminescence detection

The determination of indomethacin (INM) in pharmaceutical and biological samples by means of high-performance liquid chromatography (HPLC) with in situ electrogenerated Mn(III) chemiluminescence (CL) detection was proposed. The method was based on the direct CL reaction of INM and Mn(III), which was in situ electrogenerated by constant current electrolysis. The chromatographic separation was carried out on Nucleosil RP-C₁₈ column (250 mm × 4.6 mm; i.d., 5 μm; pore size, 100 Å) at 20 °C. The mobile phase consisted of methanol:water:acetic acid = 67:33:0.1 solution. At a flow rate of 1.0 mL min⁻¹, the total run time was 10 min. The effects of several parameters on the HPLC resolution and CL emission were studied systematically. Under the optimal conditions, a linear range from 0.01 to 10 μg mL⁻¹(R² = 0.9991), and a detection limit of 8 ng mL⁻¹ (signal-to-noise ratio = 3) for INM were achieved. The relative standard deviations (R.S.D.) for 0.1 μg mL⁻¹ INM were 2.2% within a day (n = 11) and 3.0% on 5 consecutive days (n = 6), respectively. The recovery of INM from urine samples was more than 92%. The applicability of the method for the analysis of pharmaceutical and biological samples was examined.