title: Development and validation of a high-performance thin-layer chromatographic method for determination of ofloxacin residues on pharmaceutical equipment surfaces.
abstract: An HPTLC method with densitometric quantification using fluorescence at 313 nm was developed and validated for the determination of ofloxacin residue in controlling pharmaceutical equipment cleanliness. Simulated samples at a residue level of 1 mg/m2 were prepared by spreading the calculated amount of ofloxacin solution on 1, 5, and 10 dm2 stainless steel surfaces. After evaporation of the solvent, the residue was removed by two ethanol wetted cotton swabs, which were thereafter extracted with the mixture of ethanol and Na2EDTA-water solution at pH 11 for 15 min with sonication. The extract and standards were applied on HPTLC silica gel 60 plates and then developed in a horizontal developing chamber from both sides using ethanol-conc. ammonia (4+1, v/v) as the mobile phase. The mean recovery (n=6) at 1 mg/m2 from 1, 5, and 10 dm2 was 95.3, 88.6, and 89.7% with the CV values 3.78, 4.41, and 4.97%, respectively. The absolute detection limit was 0.6 ng and the quantitation limit was 2 ng, but it was shown that these can be improved by immersion of the developed plate into a solution of liquid paraffin-n-hexane (1+2, v/v) to approximately 0.25 and 0.9 ng, respectively. The LOD of the method using detection without paraffin-n-hexane was 3, 0.6, and 0.3 microg/m2 by swabbing 1, 5, and 10 dm2, respectively. The method can be applied to routine control of pharmaceutical equipment cleanliness by sampling from stainless steel surface areas of 1 to 10 dm2 with acceptable residue limit/surface of 1 mg/m2.
authors.affiliation: National Institute of Chemistry, Laboratory for Food Chemistry, Hajdrihova 19, SI-1000 Ljubljana, Slovenia. irena.vovk@ki.si
authors.forename: Irena
authors.initials: I
authors.lastname: Vovk
authors.forename: Breda
authors.initials: B
authors.lastname: Simonovska
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry*; D002119:Calcium Carbonate / Q000737:chemistry; D002855:Chromatography, Thin Layer / Q000379:methods*; D002951:Citrates / Q000737:chemistry; D004338:Drug Combinations; D004355:Drug Stability; D004866:Equipment Contamination; D008277:Magnesium Oxide / Q000737:chemistry; D015394:Molecular Structure; D015242:Ofloxacin / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012997:Solvents
chemical_list: D000900:Anti-Bacterial Agents; D002951:Citrates; D004338:Drug Combinations; D012997:Solvents; C050615:Suby's G solution; D008277:Magnesium Oxide; D015242:Ofloxacin; D002119:Calcium Carbonate