An eco-friendly first and second derivative synchronous spectrofluorimetry for quantification of florfenicol in presence of its different degradation products. Application to kinetic stability study.

Two rapid, simple, sensitive and selective derivative spectrofluorimetric methods (first and second derivative synchronous spectrofluorimetric (FDSFS and SDSFS) procedures) have been developed for the analysis of florfenicol in the presence of its various degradation products. FDSFS was applied to assay the drug in the presence of its alkaline, oxidative and photolytic degradation products while SDSFS was used to quantify it in the presence of its acidic degradation product. These methods permitted quantification of florfenicol at corresponding λ Em of 288, 287, 279 and 284 nm without interferences from any of its degradation products. Full validation procedures were applied to the suggested method according to International Conference of Harmonization guidelines. Moreover, different degradation kinetic parameters were calculated such as half-life (t 1/2), degradation rate constant (K) and activation energy (E a). Using the analytical eco-scale, green analytical procedure index and analytical greenness metric approach AGREE as greenness assessment tools, the proposed method was found to be environmentally friendly.

Tracing the influence of caffeine on the pharmacokinetic parameters of three headache relieving pharmaceuticals applying synchronous fluorescence spectroscopy.

A simple, sensitive, and selective first derivative synchronous fluorimetric method was developed and optimized to track the influence of caffeine content in beverages on the pharmacokinetic parameters of three pharmaceuticals used in relieving headache namely, aspirin (ASP), ibuprofen (IBU), and ergotamine tartrate (ERG). A full validation procedure was carried out to impart validity to the proposed method to apply it to biological fluids. The unique dissolving power of micellar solutions was utilized to avoid multiple extraction steps for both thein vitroandin vivoexperiments, aiming to obtain acceptable recoveries and to accomplish sustainability, where 0.1 M sodium dodecyl sulphate (SDS) was used for this purpose. Moreover, the developed bioanalytical method was subjected to full validation to avoid interferences emerging from biological matrices. The greenness of the proposed method was assessed according to the Analytical Eco-Scale and proved to be excellent green carrying a score of 98%.

Simultaneous determination of ergotamine, caffeine and dipyrone in their ternary mixture by applying double divisor and first derivative ratio spectra methods.

Two validated methods namely, double divisor ratio spectra derivative spectroscopy and derivative ratio spectroscopy with zero crossing point were applied to assay a ternary mixture of ergotamine tartrate (EGT), caffeine (CAF) and dipyrone sodium (DIP) without any additional separation steps. The linearity ranges using both methods were (1.0µg/mL-70.0µg/mL), (60.0µg/mL-100.0µg/mL) and (100.0µg/mL-300.0µg/mL) for EGT, CAF and DIP respectively. Double divisor ratio spectroscopy (method A) depends on dividing the different peak responses of EGT on (summation of peaks responses of CAF and DIP each of 10.0µg/mL concentration) at λ max=342nm, 310nm and 315nm for EGT, CAF and DIP respectively. Derivative ratio spectroscopy with zero crossing point (method B) depends on dividing the peak responses of two drugs (EGT and CAF) on (10.0µg/mL of DIP) and dividing the peak response of DIP on peak response of (10.0µg/mL of EGT). The detection limits of the studied drugs applying method A were (3.54, 12.96 and 8.748µg/mL), with quantitation limits of (10.73, 39.28 and 26.51µg/mL) for EGT, CAF and DIP respectively. Regarding method B, the limits of detection and quantitation for EGT were 0.604µg/mL and 1.829µg/mL respectively: with corresponding values of 19.44µg/mL and 58.92µg/mL for CAF and 20.44µg/mL and 61.9µg/mL for DIP. The obtained results were compared to those obtained by published methods and were found to be in accordance.

Studying the suitability of hybrid micelle liquid chromatography for estimating the lipophilicity of some partial dopamine agonists used to attain the reward circuit.

Three micellar-based mobile phases were developed and optimized for the simultaneous determination of certain partial dopamine agonists that are used to overcome the withdrawal symptoms of abused drugs, namely aripiprazole, pramipexole and piribedil. The studied drugs were separated using micellar liquid chromatography, hybrid micellar liquid chromatography (HMLC) and microemulsion liquid chromatography (MELC). The three developed mobile phases were studied to estimate their suitability for the measurement of log p-values of the studied drugs. Experimental determination of log Pm/w values using the three mobile phases demonstrates that HMLC is the mobile phase of choice since the obtained practical log Pm/w values were in accordance with the reported log P values, and calculated log P and log D values. An explanation of the obtained results was presented based on the separation retention mechanism for each chromatographic technique. Furthermore, the effect of the pH and the column temperature in HMLC on the practical log Pm/w values was studied. To verify its suitability for experimental measurement of log Pm/w , HMLC was subjected to full validation according to the United States Pharmacopeia.

Analysis of clozapine in its tablets using two novel spectrophotometric reactions targeting its tertiary amino group.

Two simple spectrophotometric methodologies have been proposed and validated for the measurement of an atypical antipsychotic drug Clozapine (CLZ). Method A depends on interaction of CLZ with N-bromosuccinimide(NBS) resulting in formation of a yellowish orange colored product, measured at 320 nm. The linearity range was 5.0-70.0 µg/mL. Method B depends on condensation of the same drug with acetic acid mixed anhydride reagent producing a purple colored product, measured at 319 nm. The linearity range was 8.0-24.0 µg/mL. All parameters affecting the reaction condition (volume of both reagent, temperature, time and the different diluting solvents) were optimized. Both methods were successfully applied to assay CLZ in its pure form and tablets giving mean percentage recoveries of (98.87 ± 1.8 and 100 ± 1.7) for method A, and corresponding values of (98.6 ± 0.96 and 99.5 ± 1) for method B. Besides, the study of reactions stoichiometry was performed and the reaction mechanisms were proposed.

Micellar liquid chromatographic method for the simultaneous determination of citalopram hydrobromide with its two demethylated metabolites. Utility as a diagnostic tool in forensic toxicology.

A micellar liquid chromatographic method has been developed for the simultaneous determination of citalopram hydrobromide (CTA) with its two demethylated metabolites namely; desmethyl citalopram hydrochloride (DCTA) and didesmethyl citalopram tartrate (DDCTA). Separation was conducted on a C18 column using a mobile phase composed of 0.18 M sodium dodecyl sulphate (SDS), 15% 1-propanol, 0.3% tri-ethylamine, adjusted to pH 4 with 0.2 M o-phosphoric acid and adopting UV detection at 240 nm. Analysis was performed at 60 °C applying a flow rate of 2 mL/min. The proposed method was linear over the concentration ranges of 1.0-200.0, 0.6-200.0, and 0.5-200.0 µg/mL for CTA, DCTA, and DDCTA respectively, with corresponding limits of detection (LOD) of 0.5, 0.4, and 0.3 µg/mL and limits of quantification (LOQ) of 0.8, 0.5, and 0.4 µg/mL. The method was fully validated which allowed its application for the determination of CTA in its tablets. Moreover, the proposed method was extended to assay CTA with its metabolites in rat tissue organs samples which allowed the method to be used as a diagnostic tool in forensic toxicology.

Analytical method development and validation of spectrofluorimetric and spectrophotometric determination of some antimicrobial drugs in their pharmaceuticals.

In this study, three novel, sensitive, simple and validated spectrophotometric and spectrofluorimetric methods have been proposed for estimation of some important antimicrobial drugs. The first two methods have been proposed for estimation of two important third-generation cephalosporin antibiotics namely, cefixime and cefdinir. Both methods were based on condensation of the primary amino group of the studied drugs with acetyl acetone and formaldehyde in acidic medium. The resulting products were measured by spectrophotometric (Method I) and spectrofluorimetric (Method II) tools. Regarding method I, the absorbance was measured at 315nm and 403nm with linearity ranges of 5.0-140.0 and 10.0-100.0µg/mL for cefixime and cefdinir, respectively. Meanwhile in method II, the produced fluorophore was measured at λem 488nm or 491nm after excitation at λex 410nm with linearity ranges of 0.20-10.0 and 0.20-36.0µg/mL for cefixime and cefdinir, respectively. On the other hand, method III was devoted to estimate nifuroxazide spectrofluorimetrically depending on formation of highly fluorescent product upon reduction of the studied drug with Zinc powder in acidic medium. Measurement of the fluorescent product was carried out at λem 335nm following excitation at λex 255nm with linearity range of 0.05 to 1.6µg/mL. The developed methods were subjected to detailed validation procedure, moreover they were used for the estimation of the concerned drugs in their pharmaceuticals. It was found that there is a good agreement between the obtained results and those obtained by the reported methods.

Simultaneous Liquid Chromatographic Determination of Ebastine with Two Sympathomimetic Drugs Using a Monolithic Column.

Ebastine (EBS) has been assayed in its laboratory-prepared co-formulated tablets with either pseudoephedrine hydrochloride (PSU) or phenylephrine hydrochloride (PHR) using isocratic reversed-phase chromatography. Separation was conducted using a 50 mm × 4.6 mm i.d., Chromolith® SpeedROD RP-18 end-capped column at ambient temperature. A mobile phase composed of water:acetonitrile in a ratio of 25:75 having a pH of 3.2, has been utilized at 1 mL/min with UV detection at 254 nm for both EBS and PSU and 274 nm for PHR which in turn increased the sensitivity of the proposed method significantly. Symmetric well-separated peaks resulted in a short chromatographic run; <5 min. The proposed method was subjected to detailed validation procedures and proved to be highly sensitive as shown from limit of quantification values which were 4.7, 39.4 and 10.2 µg/mL for EBS, PSU and PHR, respectively. The proposed method was used to analyze EBS in its laboratory-prepared co-formulated tablets; the obtained results were comparable to those resulting from the reference method.

Novel validated spectrofluorimetric methods for the determination of taurine in energy drinks and human urine.

Two sensitive, selective, economic and validated spectrofluorimetric methods were developed for the determination of taurine in energy drinks and spiked human urine. Method Ι is based on fluorimetric determination of the amino acid through its reaction with Hantzsch reagent to form a highly fluorescent product measured at 490 nm after excitation at 419 nm. Method ΙΙ is based on the reaction of taurine with tetracyanoethylene yielding a fluorescent charge transfer complex, which was measured at λex /em of (360 nm/450 nm). The proposed methods were subjected to detailed validation procedures, and were statistically compared with the reference method, where the results obtained were in good agreement. Method Ι was further applied to determine taurine in energy drinks and spiked human urine giving promising results. Moreover, the stoichiometry of the reactions was studied, and reaction mechanisms were postulated.

Liquid chromatography coupled with fluorimetric detection and third derivative synchronous fluorescence spectroscopy as two analytical methods for the simultaneous determination of rabeprazole sodium and domperidone after derivatization with 4-chloro-7-nitrobenzofurazan.

Two simple, sensitive, rapid, economic and validated methods, namely reversed phase liquid chromatography (method Ι) and third derivative synchronous fluorescence spectroscopy (method ΙΙ) have been developed for the simultaneous determination of rabeprazole sodium and domperidone in their laboratory prepared mixture after derivatization with 4-Chloro-7-nitrobenzofurazan. Reversed phase chromatography was conducted using a Zorbax® SB-Phenyl column (250.0 mm × 4.6 mm id) combined with a guard column at ambient temperature with fluorimetric detection at 540 nm after excitation at 483 nm. A mobile phase composed of a mixture of distilled water with methanol and acetonitrile in a ratio of 50:20:30 adjusted pH to 4 has been used at a flow rate of 1 mL/min. Sharp well resolved peaks were obtained for domperidone and rabeprazole sodium with retention times of 5.5 and 6.4 min respectively. While in method ΙΙ, the third-derivative spectra were estimated at 507 and 436 nm for rabeprazole sodium and domperidone respectively. Linearity ranges for rabeprazole sodium and domperidone respectively in both methods were found to be 0.15-2.0 and 0.1-1.5 µg/mL. The proposed methods were successfully applied for the analysis of the two compounds in their binary mixtures, and laboratory prepared tablets. The obtained results were favorably compared with those obtained by the comparison method. Furthermore, detailed validation procedure was also conducted.

Liquid chromatographic determination of roxithromycin: application to stability studies.

A simple, stability-indicating, reversed-phase liquid chromatographic method has been developed for the determination of roxithromycin in the presence of its forced alkaline, oxidative and ultraviolet degradation products. Reversed-phase chromatography was conducted using an ODS C18 (150 × 4.6 mm i.d.) column at ambient temperature with ultraviolet detection at 215 nm. A mobile phase consisting of 0.03 M potassium dihydrogen phosphate buffer-methanol (40:60, v/v) adjusted to pH 4.5 was used for the separation of the studied drug and its degradation products at a flow rate of 1 mL/min. The method showed good linearity over the concentration range of 10.0-150.0 µg/mL with a detection limit of 2.5 µg/mL and quantification limit of 8.4 µg/mL. The proposed method was successfully applied for the analysis of roxithromycin in its commercial tablets; the obtained results were favorable compared with those obtained by the official method. Furthermore, content uniformity testing of the studied tablets was also conducted. The method was also utilized to investigate the kinetics of the different degradation products of the drug. The first-order rate constant, half-life time and activation energy of the degradation reactions were calculated.

Validated Stability indicating Spectrophotometric Method for the Determination of Acetazolamide in Dosage Forms through Complex Formation with Palladium (II).

A simple and sensitive spectrophotometric method was developed for the determination of acetazolamide (ACM) in pure form and pharmaceutical preparations. The proposed method is based on the complex formation of acetazolamide with Palladium (II) chloride in acetate buffer pH5.4 and measuring the absorbance at 308 nm. The absorbance- concentration plot was rectilinear over the concentration range of 5-70 µg/ml with a minimum detection limit (LOD) of 0.98 µg/ml, limit of quantification (LOQ) of 2.96 µg/ml, and a molar absorptivity ζ=2.7 × 10(3) L/mol.cm. The factors affecting the absorbance of the formed complex were carefully studied and optimized. The composition of the complex as well as its stability constant was also investigated. The proposed method was applied for the determination of acetazolamide in its tablets and the results obtained were favorably compared with those obtained using the official method. A proposal of the reaction pathway was postulated.

Spectrofluorimetric Determination of Famotidine in Pharmaceutical Preparations and Biological Fluids through Ternary Complex Formation with Some Lanthanide Ions: Application to Stability Studies.

A simple, sensitive and specific method was developed for the determination of famotidine (FMT) in pharmaceutical preparations and biological fluids. The proposed method is based on ternary complex formation of famotidine (FMT) with EDTA and terbium chloride TbCl3 in acetate buffer of pH 4. Alternatively, the complex is formed via the reaction with hexamine and either lanthanum chloride LaCl3, or cerous chloride CeCl3 in borate buffer of pH6.2 and 7.2 respectively. In all cases, the relative fluorescence intensity of the formed complexes was measured at 580 nm after excitation at 290 nm. The fluorescence intensity - concentration plots were rectilinear over the concentration range of 10-100, 5-70, and 5-60 ng/ml, with minimum quantification limits (LOQ) of 2.4, 2.2, and 5.2 ng/ml, and minimum limits of detection (LOD) of 0.79, 0.74, and 1.7 ng/ml upon using TbCl3, LaCl3, and CeCl3 respectively. The proposed method was applied successfully for the analysis of famotidine in dosage forms and in human plasma. The kinetics of both alkaline and oxidative induced degradation of the drug was studied using the proposed method. The apparent first order rate constant and half life time were calculated. A proposal of the reaction pathways is presented.