Quantitation of nimesulide in human plasma by high-performance liquid chromatography with ultraviolet absorbance detection and its application to a bioequivalence study.
A rapid and simple high-performance liquid chromatography-reversed phase (HPLC-RV) method with ultraviolet detection (258 nm) was developed and validated for the quantitation of nimesulide (CAS 51803-78-2) in human plasma. After the plasma samples were extracted with 6.0 ml of dichloromethane containing the internal standard phenacetin 2 microg/ml in methanol, the analysis of the nimesulide level in the plasma samples was carried out using a reverse phase Supelcosil LC-18 (15 cm x 4.6 mm x 5 pm) column. The chromatographic separation was accomplished with an isocratic mobile phase consisting of a mixture of methanolphosphate buffer (pH 3.5; 0.01 mol/l) (55:45, v/v). The inter-assay accuracy ranged from 103.4 to 113.2%, while intra-day ranged from 105.6 to 117.5%. The inter-assay precision ranged from 11.7 to 14.6%, while intra-assay ranged from 3.2 to 9.5%. The recovery of nimesulide was determined as part of the assay validation process and was excellent. The linearity of the nimesulide curves ranged from 0.20 to 15.0 microg/ml (y = 0.3857-0.0081, r = 0.9975). Short-term stability showed that nimesulide is stable in plasma for at least 24 h at room temperature, while long-term stability studies showed that nimesulide is stable in plasma for at least 180 days when stored at -20 degrees C. This validated method was successfully applied to the bioequivalence study of nimesulide in tablets in healthy volunteers.
Rolim
Clarice Madalena Bueno
CM
Departamento de Farmácia Industrial, Centro de Ciências da Saúde, Universidade Federal de Santa Maria, Santa Maria, Brazil. crolim@smail.ufsm.br
Porta
Valentina
V
Storpirtis
Sílvia
S
D000328:Adult; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000097:blood* / Q000493:pharmacokinetics*; D019540:Area Under Curve; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D018592:Cross-Over Studies; D005260:Female; D006801:Humans; D007202:Indicators and Reagents; D008297:Male; D010615:Phenacetin / Q000097:blood; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D013449:Sulfonamides / Q000097:blood* / Q000493:pharmacokinetics*; D013810:Therapeutic Equivalency
D000894:Anti-Inflammatory Agents, Non-Steroidal; D007202:Indicators and Reagents; D013449:Sulfonamides; D010615:Phenacetin; C012655:nimesulide