Determination of adapalene in gel formulation by conventional and derivative synchronous fluorimetric approaches. Application to stability studies and in vitro diffusion test.

BACKGROUND: Adapalene is a retinoid analogue with actions similar to those of tretinoin. It is used in topical treatment of mild to moderate acne. A survey of the literature reveals that no spectrofluorimetric method has been reported yet for determination of ADP, so it was thought necessary to develop a highly sensitive stability indicating spectrofluorimetric method. RESULTS: Two highly sensitive spectrofluorimetric approaches were conducted for the assay of adapalene (ADP) in its gel. In the first approach, ADP exhibits an intense native fluorescence at 389 nm after excitation at 312 nm using borate buffer (pH 7.0)/ethanol system. This approach was successfully applied for routine analysis of ADP in its gel and ideally suited to the in vitro diffusion test. To elucidate the inherent stability of ADP, bulk sample was subjected to different stress conditions as specified by ICH guidelines. The acidic and oxidative degradation products were resolved from the intact drug using second and first derivative synchronous fluorimetry at 346 and 312.45 nm, respectively (the second approach). The synchronous fluorescence was scanned at Δ λ of 80 nm in case of acidic degradation and at Δ λ of 100 nm in case of oxidative degradation. Good linearity was obtained for ADP over the range 2.0-14.0 ng/mL with good correlation coefficient   0.999 in each approach. The approaches were carefully examined in terms of linearity, accuracy and precision. They were suitable for routine quality control laboratory. Moreover, the stability-indicating power of the second approach was ascertained via forced degradation studies. CONCLUSIONS: The proposed approaches were validated and successfully applied for the quantitative assay of a small concentration of ADP in its pharmaceutical gel. The conventional spectrofluorimetry was ideally suited for in vitro diffusion test. Stability studies were also conducted using different forced degradation condition according to ICH recommendation.Graphical abstractSimultaneous determination of ADP and its degradation products.

Micellar HPLC and derivative spectrophotometric methods for the simultaneous determination of fluconazole and tinidazole in pharmaceuticals and biological fluids.

Micellar high-performance liquid chromatography (HPLC) and first-derivative ultraviolet spectrophotometry were used to simultaneously determine fluconazole (FLZ) and tinidazole (TNZ) in combined pharmaceutical dosage forms. The derivative procedure is based on the linear relationship between the drug concentration and the first derivative amplitudes at 220 and 288 nm for FLZ and TNZ, respectively. The calibration graphs were linear in the range of 1.5-9.0 µg/mL for FLZ and 10.0-60.0 µg/mL for TNZ. Furthermore, an HPLC procedure with ultraviolet detection at 210 nm was developed. For the HPLC procedure, good chromatographic separation was achieved using an ODS C18 column (250 × 4.6 mm i.d.). The mobile phase containing 0.15M sodium dodecyl sulphate, 0.3% triethylamine and 12% n-propanol in 0.02M orthophosphoric acid at pH 5.5 was pumped at a flow rate of 1 mL/min. Indapamide was used as an internal standard. The method showed good linearity over the concentration ranges of 1.5-30.0 and 10.0-200.0 µg/mL, with limits of detection of 0.36 and 2.70 µg/mL and limits of quantification of 1.1 and 8.2 µg/mL for FLZ and TNZ, respectively. The suggested methods were successfully applied for the simultaneous analysis of the drugs in their laboratory prepared mixture, co-formulated tablet and single dosage forms. Moreover the second method was also extended to the determination of the drugs in biological fluids.

Spectrofluorimetric determination of terbinafine hydrochloride and linezolid in their dosage forms and human plasma.

A highly sensitive, simple and rapid spectrofluorimetric method was developed for the determination of Terbinafine HCl (TRH) and linezolid (LNZ) in their pharmaceutical formulations. The proposed method is based on measuring the native fluorescence of the studied drugs in water at 336 nm after excitation at 275 nm for TRH and 375 nm after excitation at 254 nm for LNZ. The fluorescence-concentration plots were rectilinear over the range of 0.02-0.15 µg/mL for TRH and 0.5-5.0 µg/mL for LNZ. With lower detection limits of 3.0 and 110.0 ng/mL and a lower quantification limit of 9.0 and 320.0 ng/mL for TRH and LNZ, respectively. The method was successfully applied to the analysis of TRH in its commercial tablets, cream, gel and spray formulations and the results were in good agreement with those obtained with the official method. In addition the method was also applied to the analysis of LNZ in its capsule and I.V solution and the results were in good agreement with those obtained with the comparison method. The effect of sensitizers was studied. The method was extended to the determination of the studied drugs in spiked human plasma and the results were satisfactory.

Kinetic determination of ribavirin in drug formulations.

Two simple and sensitive kinetic methods were developed for the determination of ribavirin in bulk and in its pharmaceutical preparations using alkaline potassium permanganate as an oxidizing agent. The methods are based upon a kinetic investigation of the oxidation reaction of the drug at room temperature for fixed times of 20 and 30 minutes. In the first method, the absorbance of the colored manganate ion was measured at 610 nm, while in second method the reduction in the absorbance of permanganate was measured at 525 nm. The absorbance concentration plots were linear over the range of 3-15 µg/ml with detection limits of 0.028 µg/ml in the first method and 0.229 µg/ml for the second method. The proposed methods were applied successfully for the determination of the drug in its pharmaceutical formulations, the percentage recoveries were 100.15 ± 1.34, 100.06 ± 0.86 in the first method, and 99.60 ± 0.54, 100.43 ± 0.82 in the second method. The results obtained were compared statistically with those obtained by the official method and showed no significant differences regarding accuracy and precision.