Determination of sulindac and its metabolites in human serum by reversed-phase high-performance liquid chromatography using on-line post-column ultraviolet irradiation and fluorescence detection.
On irradiation with ultraviolet light, the antiinflammatory agent sulindac and its two metabolites sulindac sulfone and sulindac sulfide form highly fluorescent derivatives. This reaction was exploited for the sensitive and selective detection of these compounds in serum using reversed-phase high-performance liquid chromatography on a Ultrasphere octylsilane column (150 x 4.6 mm I.D.) at ambient temperature with a flow-rate of 0.5 ml/min. The analytes of interest were isolated from serum using a Bond-Elut C2 column with satisfactory recovery and selectivity. The detection limits were 10 ng/ml for each of the three analytes using 1 ml of serum and the limit of quantitation was 50 ng/ml. Linear calibration curves from 50 to 1000 ng/ml for all three analytes show coefficients of determination of 0.9999. The post-column ultraviolet irradiation was optimized and the effect of irradiation time on the fluorescence response was determined for all three analytes. Precision and accuracy of the method were 0.4-5.6 and 1.6-4.5% for sulindac, 2.3-5.6 and 1.4-5.3% for sulindac sulfone and 2.5-4.3 and 0.8-2.8% for sulindac sulfide, respectively.
Siluveru
M
M
Department of Medicinal Chemistry, College of Pharmacy, University of Georgia, Athens 30602-2352, USA.
Stewart
J T
JT
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000097:blood* / Q000378:metabolism; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D005453:Fluorescence; D006801:Humans; D007213:Indomethacin / Q000032:analysis; D016014:Linear Models; D010777:Photochemistry; D015203:Reproducibility of Results; D013467:Sulindac / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood* / Q000378:metabolism; D014466:Ultraviolet Rays
D000894:Anti-Inflammatory Agents, Non-Steroidal; D013467:Sulindac; C025462:sulindac sulfide; C025463:sulindac sulfone; D007213:Indomethacin