Environmentally benign first derivative synchronous spectrofluorimetry for the analysis of two binary mixtures containing duloxetine with avanafil or tadalafil in spiked plasma samples.

Antidepressants can cause sexual dysfunction side effects, necessitating the co-administration of phosphodiesterase type 5 inhibitors. The simultaneous determination of these drugs in biological fluids is critical for therapeutic drug monitoring. For the first time, two binary mixtures containing duloxetine with either avanafil or tadalafil were estimated utilizing simple green spectrofluorimetric methods without the need for a previous separation step. The study was based on first derivative synchronous spectrofluorimetry in ethanol using a change in wavelength difference (∆λ) of 20 and 25 nm for the first and second combinations, respectively. Duloxetine and avanafil were estimated at 297.7 and 331 nm in their binary mixture, while duloxetine and tadalafil were determined at 290.3 and 297.7 nm, respectively. The linearity was achieved over the ranges of 0.1-1.5 µg mL-1 for both duloxetine and avanafil and 0.01-0.40 µg mL-1 for tadalafil, with limits of detection of 0.013, 0.022, and 0.004 µg mL-1 for duloxetine, avanafil, and tadalafil, respectively. Successful application of the developed approaches was accomplished for the estimation of the two mixtures in dosage forms as well as human plasma with excellent percentage recoveries (96-103.75% in plasma), which supports their suitability for use in quality control laboratories and pharmacokinetic studies. Moreover, the adopted approaches' greenness was evidenced by applying three tools.

Green, Validated HPLC Method Coupled With Fluorescence Detection for the Analysis of Tafluprost in its Pure Form and Ophthalmic Formulation: Application to Aqueous Humor and Content Uniformity Testing.

A green, simple and sensitive HPLC method coupled with fluorescence detection was implemented for the quantitative determination of the anti-glaucoma drug tafluprost (TFL). Liquid chromatography was performed on HyperClone™ ODS (C18) column of dimensions; 150 × 4.6 mm i.d. and 5 µm particle size using a green eluent; ethanol:0.01 M phosphate buffer (60:40 v/v, pH 4.5) delivered at 1 mL min-1. Fluorescence detection was accomplished at 220 nm (excitation) and 292 nm (emission). Bimatoprost (BIM) was used as an internal standard (I.S.). In this method, TFL was eluted after 6.70 minutes. The method satisfied International Council for Harmonization (ICH) validation guidelines, as proved by good linearity (r = 0.9999, over the range 0.05-2 µg mL-1), accuracy (recovery average 100.13 ± 1.27%), precision, robustness and specificity. The limit of detection and limit of quantification were found to be 0.016 and 0.048 µg mL-1, respectively. The proposed method has been successfully applied for the estimation of TFL in eye drops and aqueous humor. For the first time, the approach was applied with acceptable results for the evaluation of the uniformity of TFL eye drops content. Furthermore, Green Analytical Procedure Index (GAPI) and analytical Eco-scale were used to prove that the proposed HPLC method is environmentally friendly.

Factorial design-assisted reverse phase HPLC-UV approach for the concurrent estimation of cetirizine and azelastine in aqueous humor.

A new analytical quality by design-assisted HPLC-UV approach is presented, for the first time, for the concurrent determination of cetirizine (CTZ) and azelastine (AZE) in raw materials, commercial eye drops and aqueous humor. The two drugs are co-administered as eye drops in severe ocular allergies. A 23 full factorial design was adopted for the chromatographic optimization to ensure the best analytical performance and reliability, as well as to save time, effort and solvent consumption. The parameters, including pH, acetonitrile ratio, and flow rate, were selected as independent factors. The responses analyzed were resolution and tailing of peaks. The separation was achieved through isocratic elution on C8 column with mobile phase made up of acetonitrile: 0.3% triethylamine of pH 5 (60:40 v/v) at a flow rate of 1.2 mL min-1 and detection at 216 nm. The elution time was less than 6 min. The approach was fully validated in accordance with International Council for Harmonization (ICH) guidelines. Good linearity was achieved over the concentration ranges of 1.0-30 and 0.5-10 µg mL-1 with limits of detection of 0.310 and 0.158 µg mL-1 and limits of quantification of 0.940 and 0.479 µg mL-1 for CTZ and AZE, respectively, with correlation coefficients of 0.9998. The intra- and inter-day precisions were lower than 2%. The good sensitivity of the approach permits the analysis of CTZ and AZE in spiked aqueous humor with mean percentage recoveries of 100.93 ± 1.42 and 100.11 ± 1.55, respectively. The statistical comparison between results of the developed method and the comparison method revealed no differences, indicating the accuracy of the method.

Green HPLC method with time programming for the determination of the co-formulated eye drops of tafluprost and timolol in their challengeable ratio.

A new, simple and selective HPLC method was implemented for the simultaneous estimation of tafluprost (TFL) and timolol (TIM) in their new anti-glaucoma combination in the challengeable ratio of 3 and 1000 for TFL and TIM, respectively. Separation was achieved using a BDS Hypersil phenyl column and a mobile phase made up of acetonitrile: 0.015 M phosphate buffer (50:50 v/v, pH 3.5) delivered at 1 mL min-1 and the separation was completed in less than 6 min. UV detection was time programmed at 220 nm for the first 4.5 min and later at 254 nm. Mebeverine (MEB) was used as an internal standard (I.S.). The linearity was observed in the ranges of 0.6-45 and 50-2000 µg mL-1 with limits of detection (LOD) of 0.18, 16.48 µg mL-1 and limits of quantification (LOQ) of 0.55, 49.94 µg mL-1 for TFL and TIM, respectively. The method satisfied International Council for Harmonization (ICH) validation guidelines. The study was extended to the estimation of the studied drugs in their co-formulated eye drops as well as in their single dosage forms with acceptable percentage recoveries. Moreover, Green Analytical Procedure Index (GAPI) and analytical Eco-scale were investigated to confirm the greenness of the proposed HPLC method.

Synchronous Fluorescence as a Green and Selective Method for the Simultaneous Determination of Cetirizine and Azelastine in Aqueous Humor.

A green, simple, quick and economical method is implemented for the first time for the simultaneous estimation of cetirizine (CTZ) and azelastine (AZE) as co-administered eye drops. The method relies on synchronous spectrofluorimetry with ∆λ = 60 nm. Cetirizine can be estimated at 231 nm and AZE can be measured at 294 nm, each at the other's zero crossing point. All factors affecting the method were studied and properly optimized. Good correlation was obtained in the range of 0.1-2 µg mL-1 for both drugs. The limits of detection were 0.014 and 0.010 µg mL-1 and limits of quantitation were 0.043 and 0.029 µg mL-1 for CTZ and AZE, respectively. Moreover, ICH guidelines were carried out to validate the adopted method. The method was suitable for the analysis of CTZ and AZE in synthetic mixtures, eye drops and aqueous humor. The mean percentage of recoveries of CTZ and AZE in spiked aqueous humor were 99.83 and 99.37, respectively. Furthermore, Green Analytical Procedure Index (GAPI) and analytical Eco-scale approaches were used to evaluate the greenness of the suggested method.

Green highly sensitive spectrofluorimetric method for rapid determination of tafluprost in its pure form and ophthalmic formulation.

A green, simple and easy spectrofluorimetric method was studied for rapid estimation of tafluprost (TFL). The native fluorescence of TFL was measured at 292 nm after excitation at 220 nm. The results were linear in water over the concentration range 50-600 ng ml-1 with a correlation coefficient r = 0.9999 and intercept 1.1555. The limit of detection and limit of quantification were found to be 7.87 and 23.86 ng ml-1 , respectively. Neither different pH nor surfactants enhanced the fluorescence intensity. The high sensitivity of this spectrofluorimetric method makes it suitable for analysis of low concentrations of tafluprost in commercially available ophthalmic formulations. This procedure was validated according to International Council for Harmonisation Guidelines.