Spectrofluorimetric methods for the determination of mirabegron by quenching tyrosine and L-tryptophan fluorophores: Recognition of quenching mechanism by stern volmer relationship, evaluation of binding constants and binding sites.

Green spectrofluorimetric methods have been adopted for the determination of Mirabegron (MG) in pure drug and pharmaceutical dosage form. The developed methods based on fluorescence quenching of tyrosine and L-tryptophan amino acids fluorophores by the effect of Mirabegron as a quencher. Experimental conditions of the reaction were studied and optimized. The Fluorescence quenching (ΔF) values were proportional to the concentration range of MG 2-20 µg/ml for the tyrosine-MG system in buffered media pH 2 and 1-30 µg/ml for L-tryptophan-MG system pH 6. Good correlation coefficients with low detection limits of 0.163 and 0.234 µg/ml for the two systems respectively. Method validation was applied according to ICH guidelines. The cited methods were successively applied for MG determination in tablet formulation. No statistically significant difference between the results of the cited and the reference methods regarding t and F tests. The proposed spectrofluorimetric methods are simple, rapid, eco-friendly and can contribute to MG's methodologies in quality control labs. Stern-Volmer relationship, the effect of temperature, quenching constant (Kq), and UV spectra were studied to identify the mechanism by which the quenching might occur. The results demonstrated that fluorescence quenching of tyrosine was a dynamic quenching process and L-tryptophan was static. The double log plots were constructed to determine the binding constants and binding sites. The greenness profile of the developed methods has been assessed by Green Analytical procedure index (GAPI) and Analytical Greenness Metric Approach (AGREE).

Selective Determination of Entecavir in the Presence of its Oxidative Degradate by Spectrophotometric and Chromatographic Methods.

BACKGROUND: Entecavir (ENT) is an antiretroviral agent prescribed for the treatment of the hepatitis B virus(HBV) and human immunodeficiency virus(HIV). OBJECTIVE: Development and validation of three simple, sensitive, selective, and precise methods for determination of ENT in the presence of its oxidative degradation product (ENT deg.). METHOD: The first method was based on second derivative (D2) spectrophotometry through measuring the peak amplitude of D2 spectra at 293.6 nm. The second one is mean centering of the ratio spectra (MCR), which enabled measurement of the peak amplitude at 280.0 nm. The third method was HPLC, where ENT was separated from ENT deg. using a Zobrax C18 column and methanol:water (30:70, v/v) with pH 3 as a mobile phase. The three developed methods were validated according to the International Conference on Harmonization guidelines. RESULTS: Linearity range of ENT was 5.00-50.00 µg/mL for both D2 and MCR. However, higher sensitivity was achieved using HPLC (1.00-50.00 µg/mL). Accuracy of ENT were 100.60 ± 0.547%, 101.55 ± 1.2071%, and 100.61 ± 1.207% for D2, MCR, and HPLC methods, respectively, and precision was within 1.280. CONCLUSIONS: The developed methods were successfully applied for the determination of ENT in Tecavir® tablets without interference from ENT deg. They showed no significant difference in comparison with the official method and they can be applied in the quality analysis of ENT with high selectivity, accuracy, and precision. HIGHLIGHTS: ENT was quantified using two spectrophotometric (D2 and MCR) methods and an HPLC method in presence of ENT deg. The proposed methods were applied to analysis of ENT tablets with high selectivity, sensitivity, and accuracy.

Spectrophotometric and spectrofluorimetric methods for the determination of ramipril in its pure and dosage form.

This paper describes three sensitive spectrophotometric and spectrofluorimetric methods for determination of ramipril in its pure form and pharmaceutical tablets. The first method is based on the oxidation of the drug with 1-chlorobenzotriazole reagent (CBT) in strong alkaline medium followed by measuring the absorbance at 350 nm. The method obeys Beer's law over concentration range 15-50 microg ml(-1). For the second and third, both are non-extractive methods based on the formation of ternary complex between copper (II), eosin and ramipril in the presence of methylcellulose as surfactant. Spectrophotometrically, under the optimum condition, the ternary complex showed an absorption maximum at 543 nm. The method obeys Beer's law over concentration range of 20-80 microg ml(-1). A fluorescence quenching method for the determination of ramipril by forming this ternary complex was also investigated for the propose of enhance the sensitivity of the determination. The methods are simple, sensitive, and accurate. The results obtained are reproducible with a coefficient of variation less than 2%. The proposed have been successfully applied to the assay of ramipril in tablets. The results compare favorably with official method.

Extractive-spectrophotometric determination of disopyramide and irbesartan in their pharmaceutical formulation.

Picric acid, bromocresol green, bromothymol blue, cobalt thiocyanate and molybdenum(V) thiocyanate have been tested as spectrophotometric reagents for the determination of disopyramide and irbesartan. Reaction conditions have been optimized to obtain coloured comoplexes of higher sensitivity and longer stability. The absorbance of ion-pair complexes formed were found to increases linearity with increases in concentrations of disopyramide and irbesartan which were corroborated by correction coefficient values. The developed methods have been successfully applied for the determination of disopyramide and irbesartan in bulk drugs and pharmaceutical formulations. The common excipients and additives did not interfere in their determination. The results obtained by the proposed methods have been statistically compared by means of student t-test and by the variance ratio F-test. The validity was assessed by applying the standard addition technique. The results were compared statistically with the official or reference methods showing a good agreement with high precision and accuracy.

Spectrophotometric and spectrofluorimetric methods for analysis of acyclovir and acebutolol hydrochloride.

Simple and sensitive spectrophotometric and spectrofluorimetric methods are described for analysis of acyclovir and acebutolol hydrochloride. The proposed methods are based on oxidation of the selected drugs with cerium(IV) ion in acidic medium with subsequent measurement of either the decrease in absorbance at 320nm or the fluorescence intensity of the produced cerous(III) ion at 361-363nm (excitation at 250nm). Beer's law obeyed from 2 to 8, 0.25 to 2.5microgcm-1 acyclovir, 1 to 7 and 0.25 to 2.5microgml-1 acebutolol hydrochloride, using the spectrophotometric and spectrofluorimetric method, respectively. The proposed method were successfully applied for determination of the selected drugs in their pharmaceutical preparations with good recoveries.

Spectrophotometric and spectrodensitometric determination of paracetamol and drotaverine HCl in combination.

Thin-layer chromatography, first derivative, ratio spectra derivative spectrophotometry and Vierordt's method have been developed for the simultaneous determination of paracetamol and drotaverine HCl. TLC densitometric method depends on the difference in Rf values using ethyl acetate:methanol:ammonia (100:1:5 v/v/v) as a mobile phase. The spots of the two drugs were scanned at 249 and 308 nm over concentration ranges of 60-1200 microg/ml and 20-400 microg/ml with mean percentage recovery 100.11%+/-1.91 and 100.15%+/-1.87, respectively. The first derivative spectrophotometric method deals with the measurements at zero-crossing points 259 and 325 nm with mean percentage recovery 99.25%+/-1.08 and 99.45%+/-1.14, respectively. The ratio spectra first derivative technique was used at 246 and 305 nm with mean percentage recovery 99.75%+/-1.93 and 99.08%+/-1.22, respectively. Beer's law for first derivative and ratio spectra derivative methods was obeyed in the concentration range 0.8-12.8 and 0.4-6.4 microg/ml of paracetamol and drotaverine HCl, respectively. Vierordt's method was applied to over come the overlapping of paracetamol and drotaverine HCl in zero-order spectra in concentration range 2-26 and 2-40 microg/ml respectively. The suggested methods were successfully applied for the analysis of the two drugs in laboratory prepared mixtures and their pharmaceutical formulation. The validity of the methods was assessed by applying the standard addition technique. The obtained results were statistically agreed with those obtained by the reported method.

Spectrophotometric and spectrofluorimetric methods for analysis of acyclovir and acebutolol hydrochloride.

Simple and sensitive spectrophotometric and spectrofluorimetric methods are described for analysis of acebutolol hydrochloride. The proposed methods are based on oxidation of the selected drug with cerium(IV) ion in acidic medium with subsequent measurement of either the decrease in absorbance at 320 nm or the fluorescence intensity of the produced cerous(III) ion at 363 nm (excitation at 250 nm). Beer's law obeyed from 1.0-7.0 microg ml(-1) and 0.25-2.5 microg ml(-1) acebutolol hydrochloride, using the spectrophotometric and spectrofluorimetric method, respectively. The proposed methods were successfully applied for determination of the selected drug in its pharmaceutical preparation with good recoveries.

Spectrophotometric and spectrofluorimetric methods for analysis of tramadol, acebutolol and dothiepin in pharmaceutical preparations.

Sensitive spectrophotometric and spectrofluorimetric methods are described for the determination of tramadol, acebutolol and dothiepin (dosulepin) hydrochlorides. The two methods are based on the condensation of the cited drugs with the mixed anhydrides of malonic and acetic acids at 60 degrees C for 25-40 min. The coloured condensation products are suitable for the spectrophotometric and spectrofluorimetric determination at 329-333 and 431-434 nm (excitation at 389 nm), respectively. For the spectrophotometric method, Beer's law was obeyed from 0.5 to 2.5 microg ml(-1) for tramadol, dothiepin and 5-25 microg ml(-1) for acebutolol. Using the spectrofluorimetric method linearity ranged from 0.25 to 1.25 microg ml(-1) for tramadol, dothiepin and 1-5 microg ml(-1) for acebutolol. Mean percentage recoveries for the spectrophotometric method were 99.68+/-1.00, 99.95+/-1.11 and 99.72+/-1.01 for tramadol, acebutolol and dothiepin, respectively and for the spectrofluorimetric method, recoveries were 99.5+/-0.844, 100.32+/-0.969 and 99.82+/-1.15 for the three drugs, respectively. The optimum experimental parameters for the reaction has been studied. The validity of the described procedures was assessed. Statistical analysis of the results has been carried out revealing high accuracy and good precision. The proposed methods were successfully applied for the determination of the selected drugs in their pharmaceutical preparations with good recoveries. The procedures were accurate, simple and suitable for quality control application.

New colorimetric methods for the determination of trazodone HCl, famotidine, and diltiazem HCl in their pharmaceutical dosage forms.

Two sensitive and simple spectrophotometric methods are developed for the determination of trazodone HCl, famotidine, and diltiazem HCl in pure and pharmaceutical preparations. The methods are based on the oxidation of the cited drugs with iron(III) in acidic medium. The liberated iron(II) reacts with 1,10-phenanthroline (method A) and the ferroin complex is colorimetrically measured at 510 nm against reagent blank. Method B is based on the reaction of the liberated Fe(II) with 2,2-bipyridyl to form a stable colored complex with lambda(max )at 520 nm. Optimization of the experimental conditions was described. Beer's law was obeyed in the concentration range of 1-5, 2-12, and 12-32 microg mL(-1) for trazodone, famotidine, and diltiazem with method A, and 1-10 and 8-16 microg mL(-1) for trazodone and famotidine with method B. The apparent molar absorptivity for method A is 1.06x10(5), 2.9x10(4), 1.2x10(4) and for method B is 9.4x10(4 )and 1.6x10(4), respectively. The suggested procedures could be used for the determination of trazodone, famotidine, and diltiazem, both in pure and dosage forms without interference from common excipients.

Spectrophotometric methods for determination of enalapril and timolol in bulk and in drug formulations.

Two simple and accurate spectrophotometric methods for determination of timolol and enalapril maleate are described. The first method is based on chelate formation with palladium(II) chloride in buffered medium. The second method is based on the formation of the colored complex between palladium(II), eosin, and the two cited drugs using methylcellulose as surfactant to increase the solubility and intensity of the formed complexes. Under optimum conditions the complexes showed maximum absorption at 369.4 nm and 362.8 nm for timolol and enalapril maleate, respectively, in the first method and 552.2 and 550.6 nm for the second method. Apparent molar absorptivities were 1.8 x 10(3) and 1.3 x 10(3) and Sandell's sensitivities were 5.9 x 10(-4) and 2.7 x 10(-4) for timolol and enalapril maleate in the first method; in the second method molar absorptivities were 2.8 x 10(4) and 1.1 x 10(4) while Sandell's constants were 9.1 x 10(-3) and 2.3 x 10(-3) for timolol and enalapril maleate. The solutions of the complexes obeyed Beer's law in the concentration ranges 20-200 micro g mL(-1) and 50-300 micro g mL(-1) for timolol and enalapril maleate, respectively. In the second method, because the reaction was more sensitive the ranges were reduced to 1.6-16 micro g mL(-1) for timolol 8-56 micro g mL(-1) for enalapril maleate. The proposed methods were applied to the determination of the two drugs in their pharmaceutical formulation.

Colorimetric determination of gabapentin in pharmaceutical formulation.

Three accurate, simple and precise colorimetric methods for the determination of gabapentin in capsules are developed. The first method is based on the reaction of gabapentin with vanillin (Duquenois reagent) in the presence of McIlvain buffer pH 7.5 and the color developed was measured at 376 nm. The linearity range was found to be 80-360 microg ml(-1). The second is based on the reaction of the primary amino group of gabapentin with ninhydrin reagent in N,N'-dimethylformamide (DMF) medium producing a colored product which absorbs maximally at 569 nm. Beer's law is obeyed in the concentration range 40-280 microg ml(-1) of gabapentin. The third method is based on the reaction of gabapentin with p-benzoquinone (PBQ) to form a colored product with lambda(max) at 369 nm. The products of the reaction were stable for 2 h at 30 degrees C, shifts of the wavelength of maximum absorbance were not observed for up to 24 h after starting the reaction. The absorbance is proportional to gabapentin concentration in the range 80-320 microg ml(-1). The optimum experimental parameters for the reactions have been studied. The validity of the described procedures was assessed. Statistical analysis of the results has been carried out revealing high accuracy and good precision. The suggested procedures could be used for the determination of gabapentin in capsules. The procedures were rapid, simple and suitable for quality control application.

Kinetic spectrophotometric determination of tramadol hydrochloride in pharmaceutical formulation.

Two simple and sensitive kinetic methods for the determination of tramadol hydrochloride are described. The first method is based upon a kinetic investigation of the oxidation reaction of the drug with alkaline potassium permanganate at room temperature for a fixed time at 20 min. The absorbance of the colored manganate ions was measured at 610 nm. The second method is based on the reaction of tramadol hydrochloride with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in presence of 0.1 M sodium bicarbonate. The spectrophotometric measurements were recorded by measuring the absorbance at 467 nm, at fixed time at 25 min on thermostated water bath at 90+/-1 degrees C. All variables affecting the development of the colour have been investigated and the conditions were optimised. The absorbance concentration plots in both methods were rectilinear over the range 5-25 and 50-250 microg ml(-1), for the first and second methods, respectively. The two methods have been applied successfully to commercial capsule and ampoule dosage form. The results obtained are compared statistically with those given by the reference spectrophotometric method. The determination of tramadol hydrochloride by the fixed concentration and rate constant methods is feasible with the calibration equations obtained, but the fixed time method proves to be more applicable.

Potentiometric determination of famotidine in pharmaceutical formulations.

Two new potentiometric methods for determination of famotidine in pure form and in its pharmaceutical tablet form are developed. In the first method, the construction of plasticised poly(vinyl chloride) (PVC) matrix-type famotidine ion-selective membrane electrode and its use in the potentiometric determination of famotidine in pharmaceutical preparations are described. It is based on the use of the ion-associate species, formed by famotidine cation and tetraphenyl borate (TPB) counterion. The electrode exhibited a linear response for 1 x 10(-3)-1 x 10(-5) M of famotidine solutions over the pH range 1-5 with an average recovery of 99.26% and mean standard deviation of 1.12%. Common organic and inorganic cations showed negligible interference. In the second method, the conditions for the oxidimetric titration of famotidine have been studied. The method depends on using lead(IV) acetate for oxidation of the thioether contained in famotidine. The titration takes place in presence of catalytic quantities of potassium bromide (KBr). Direct potentiometric determination of 1.75 x 10(-2) M famotidine solution showed an average recovery of 100.51% with a mean standard deviation of 1.26%. The two methods have been applied successfully to commercial tablet. The results obtained reveal good percentage recoveries, which are in good agreement with those obtained by the official methods.

Spectrophotometric and AAS determination of ramipril and enalapril through ternary complex formation.

Two sensitive, spectrophotometric and atomic absorption spectrometric procedures are developed for the determination of two antihypertensive agents (enalapril maleate and ramipril). The spectrophotometric procedures for the two cited drugs are based on ternary complex formation. The first ternary complex (copper(II), eosin, and enalapril) was estimated by two methods; the first depends on its extraction with chloroform measuring at 533.4 nm. Beer's law was obeyed in concentration range from 56 to 112 microg ml(-1). The second method for the same complex depends on its direct measurement after addition of methylcellulose as surfactant at the pH value 5 at 558.8 nm. The concentration range is from 19 to 32 microg ml(-1). The second ternary complex (iron(III), thiocyanate, and ramipril) was extracted with methylene chloride, measuring at 436.6 nm, with a concentration range 60-132 microg ml(-1). The direct atomic absorption spectrometric method through the quantitative determination of copper or iron content of the complex was also investigated for the purpose of enhancing the sensitivity of the determination. The spectrophotometric and atomic absorption spectrometric procedures hold their accuracy and precision well when applied to the determination of ramipril and enalapril dosage forms.