Mathematical filtration of quaternary mixture to the zero order spectra of its individual components.

Simple accurate and precise validated UV spectrophotometric methods have been described here for the simultaneous determination of Tretinoin (TN) Eusolex (EX) Hydroquinone (HQ) & Hydrocortisone acetate (HC) in their dosage form. Simultaneous determination of these four drugs was a major challenge till now. Each spectrum was filtered alone to its zero-order absorption spectrum (D0) form getting spectra typical to each pure component separately. Four methods were applied depending upon constants obtained from extended regions of partially overlapped spectra in the zero or first derivative forms. The methods applied are constant multiplication coupled with spectrum subtraction method (CM-SS) derivative transformation coupled with spectrum subtraction method (DT-SS) constant value method (CV) and concentration value (conc.value) method. The partially overlapped spectra of TN and EX in the mixture were obtained by CM-SS in their zero-order form allowing direct measurement at their λmax while the resolved binary mixture of HQ and HC obtained by SS; was determined by derivatization and transformed to their zero order by DT-SS. Also Ex and HQ concentrations were determined by the graphical representation of data only without regression equation by concentration value method and the results were compared to the conventional constant value method using a regression equation. The methods applied to the quaternary mixture under study were successfully applied for the simultaneous determination of the four drugs in synthetic mixtures and in their combined dosage form Tritospot® cream. Comparing the acquired results statistically together and to official methods demonstrated no significant difference.

Three Different Approaches Based on Derivative Ratio Spectra for Spectrophotometric Resolution of a Quaternary Mixture in Semisolid Dosage Form.

BACKGROUND: Recent incorporated spectrophotometer software supporting mathematical methods was considered as an optimum key for the resolution of multicomponent mixtures. OBJECTIVE: Several spectrophotometric techniques are introduced for the determination of mixtures of tretinoin (TN), hydroquinone (HQ), and fluocinolone acetonide (FA), in the presence of the preservative methyl paraben (MP), without any separation procedure, taking into consideration the presence of two minor components and the severe overlap of their spectra. METHOD: Constant multiplication coupled spectrum subtraction resolved the quaternary mixture into the zero-order absorption spectrum of TN alone and a severely overlapped, ternary mixture of HQ, FA, and MP. Three approaches based on the derivative ratio spectra were applied to resolve this ternary, severely overlapped mixture: derivative ratio-zero-crossing point method, factorized derivative ratio method, and double divisor derivative ratio method. RESULTS: The work was conducted over a concentration range of 1-10, 4-38, and 4-35 µg/mL, for TN, HQ, and FA, respectively. The results obtained were compared statistically to each other and to the official methods, showing no significant difference. CONCLUSIONS: The proposed methods were successfully applied for the simultaneous determination of the three drugs in the presence of MP in synthetic mixtures and in their combined dosage form (Trimelasma® cream) with very good accuracy and precision. HIGHLIGHTS: This was a comparative study between conventional and new methods for resolving ternary, severely overlapped mixtures. Mathematical manipulation steps and enrichment techniques aided accurate quantification of the minor components in mixtures.

Full spectrum and selected spectrum based chemometric methods for the simultaneous determination of Cinnarizine and Dimenhydrinate in laboratory prepared mixtures and pharmaceutical dosage form.

Three chemometric methods namely, concentration residual augmented classical least squares (CRACLS), spectral residual augmented classical least squares (SRACLS) and partial least squares (PLS) were applied for the simultaneous quantitative determination of Cinnarizine and Dimenhydrinate in their binary mixtures. All techniques were applied with and without variable selection using genetic algorithm (GA) resulting in six models (CRACLS, GA-CRACLS, SRACLS, GA-SRACLS, PLS, GA-PLS). These models were applied for the simultaneous determination of the drugs in their laboratory prepared mixtures and pharmaceutical dosage form via handling their UV spectral data. It was found that GA based models are simpler and more robust than those built with the full spectral data. The proposed models were found to be simple, fast and require no preliminary separation steps; so they can be used for the routine analysis of this binary mixture in quality control laboratories.

New methods for amlodipine and valsartan native spectrofluorimetric determination, with factors optimization study.

Four native fluorescence methods were suggested for simultaneous determination of amlodipine (AML) and valsartan (VAL). These methods were based on excitation of both drugs at λ(ex) 300 nm, in one step, to give maximum emission at λ(em) 378 and 496 nm for AML and VAL, respectively. The first method, single λ(ex) method, was used without any additions. The sensitivity of this method was further increased by the addition of hydroxy propylmethyl cellulose (HPMC) surfactant, ß-cyclodextrin, or ferric oxide magnetite nanoparticles, in the other three methods. Different types of surfactants, and different concentration levels of both ß-cyclodextrin and ferric oxide nanoparticles, were scanned to determine the optimum conditions for enhancing the sensitivity. Some factors affecting the fluorescence intensity of both cited drugs, like the type and volume of the added solvent (to be used as a sensing agent), and pH of measurement were studied and optimized. The proposed methods could be used in determination of AML and VAL in bulk powder, their laboratory prepared mixtures and pharmaceutical formulations. The obtained results were statistically compared to each other and to that of some reported methods. The specificity of the developed methods was investigated, and the methods were validated according to ICH guidelines.

A comparative study of novel spectrophotometric resolution techniques applied for pharmaceutical mixtures with partially or severely overlapped spectra.

Simultaneous determination of mixtures of lidocaine hydrochloride (LH), flucortolone pivalate (FCP), in presence of chlorquinaldol (CQ) without prior separation steps was applied using either successive or progressive resolution techniques. According to the concentration of CQ the extent of overlapping changed so it can be eliminated from the mixture to get the binary mixture of LH and FCP using ratio subtraction method for partially overlapped spectra or constant value via amplitude difference followed by ratio subtraction or constant center followed by spectrum subtraction spectrum subtraction for severely overlapped spectra. Successive ratio subtraction was coupled with extended ratio subtraction, constant multiplication, derivative subtraction coupled constant multiplication, and spectrum subtraction can be applied for the analysis of partially overlapped spectra. On the other hand severely overlapped spectra can be analyzed by constant center and the novel methods namely differential dual wavelength (D(1) DWL) for CQ, ratio difference and differential derivative ratio (D(1) DR) for FCP, while LH was determined by applying constant value via amplitude difference followed by successive ratio subtraction, and successive derivative subtraction. The spectra of the cited drugs can be resolved and their concentrations are determined progressively from the same ratio spectrum using amplitude modulation method. The specificity of the developed methods was investigated by analyzing laboratory prepared mixtures and were successfully applied for the analysis of pharmaceutical formulations containing the cited drugs with no interference from additives. The proposed methods were validated according to the ICH guidelines. The obtained results were statistically compared with those of the official or reported methods; using student t-test, F-test, and one way ANOVA, showing no significant difference with respect to accuracy and precision.

Development and validation of a stability-indicating micellar liquid chromatographic method for the determination of timolol maleate in the presence of its degradation products.

A stability-indicating micellar liquid chromatographic (MLC) method was developed and validated for the quantitative determination of timolol maleate (TM) in the presence of its degradation products resulting from accelerated degradation in a run time not more than 8 min. TM was subjected to stress conditions of hydrolysis (including alkaline, acidic and thermal hydrolysis) and oxidation. An isocratic, rapid and mobile phase saving the micellar LC method was developed with a BioBasic phenyl column (150 × 1.0 mm, 5 µm particle size) and a micellar mobile phase composed of 0.1 M sodium dodecyl sulfate, 10% of 1-propanol and 0.1% of triethylamine in 0.035 M ortho-phosphoric acid. The flow rate of the mobile phase was 0.1 mL/min. UV detection was adjusted at 298 nm and performed at room temperature. The method has been validated according to the International Conference on Harmonisation guidelines. The method is successfully applied for the determination of TM in bulk powder and pharmaceutical dosage form.

Comparative study between univariate spectrophotometry and multivariate calibration as analytical tools for simultaneous quantitation of Moexipril and Hydrochlorothiazide.

Three simple, accurate, reproducible, and selective methods have been developed and subsequently validated for the simultaneous determination of Moexipril (MOX) and Hydrochlorothiazide (HCTZ) in pharmaceutical dosage form. The first method is the new extended ratio subtraction method (EXRSM) coupled to ratio subtraction method (RSM) for determination of both drugs in commercial dosage form. The second and third methods are multivariate calibration which include Principal Component Regression (PCR) and Partial Least Squares (PLSs). A detailed validation of the methods was performed following the ICH guidelines and the standard curves were found to be linear in the range of 10-60 and 2-30 for MOX and HCTZ in EXRSM method, respectively, with well accepted mean correlation coefficient for each analyte. The intra-day and inter-day precision and accuracy results were well within the acceptable limits.

Comparative study between univariate spectrophotometry and multivariate calibration as analytical tools for quantitation of Benazepril alone and in combination with Amlodipine.

Four simple, accurate, reproducible, and selective methods have been developed and subsequently validated for the determination of Benazepril (BENZ) alone and in combination with Amlodipine (AML) in pharmaceutical dosage form. The first method is pH induced difference spectrophotometry, where BENZ can be measured in presence of AML as it showed maximum absorption at 237nm and 241nm in 0.1N HCl and 0.1N NaOH, respectively, while AML has no wavelength shift in both solvents. The second method is the new Extended Ratio Subtraction Method (EXRSM) coupled to Ratio Subtraction Method (RSM) for determination of both drugs in commercial dosage form. The third and fourth methods are multivariate calibration which include Principal Component Regression (PCR) and Partial Least Squares (PLSs). A detailed validation of the methods was performed following the ICH guidelines and the standard curves were found to be linear in the range of 2-30µg/mL for BENZ in difference and extended ratio subtraction spectrophotometric method, and 5-30 for AML in EXRSM method, with well accepted mean correlation coefficient for each analyte. The intra-day and inter-day precision and accuracy results were well within the acceptable limits.

Successive spectrophotometric resolution as a novel technique for the analysis of ternary mixtures of pharmaceuticals.

A novel spectrophotometric technique was developed for the simultaneous determination of ternary mixtures, without prior separation steps. This technique was called successive spectrophotometric resolution technique. The technique was based on either the successive ratio subtraction or successive derivative subtraction. The mathematical explanation of the procedure was illustrated. In order to evaluate the applicability of the methods a model data as well as an experimental data were tested. The results from experimental data related to the simultaneous spectrophotometric determination of lidocaine hydrochloride (LH), calcium dobesilate (CD) and dexamethasone acetate (DA); in the presence of hydroquinone (HQ), the degradation product of calcium dobesilate were discussed. The proposed drugs were determined at their maxima 202 nm, 305 nm, 239 nm and 225 nm for LH, CD, DA and HQ respectively; by successive ratio subtraction coupled with constant multiplication method to obtain the zero order absorption spectra, while by applying successive derivative subtraction they were determined at their first derivative spectra at 210 nm for LH, 320 nm or P(292-320) for CD, 256 nm or P(225-252) for DA and P(220-233) for HQ respectively. The calibration curves were linear over the concentration range of 2-20 µg/mL for both LH and DA, 6-50 µg/mL for CD, and 3-40 µg/mL for HQ. The proposed methods were checked using laboratory-prepared mixtures and were successfully applied for the analysis of pharmaceutical formulation containing the cited drugs with no interference from other dosage form additives. The proposed methods were validated according to the ICH guidelines. The obtained results were statistically compared with those of the official BP methods for LH, DA, and CD, and with the official USP method for HQ; using student t-test, F-test, and one way ANOVA, showing no significant difference with respect to accuracy and precision.

Colorimetric and fluorimetric methods for determination of panthenol in cosmetic and pharmaceutical formulation.

Two methods are suggested for determination of panthenol. The first is colorimetric method where panthenol is subjected to alkaline hydrolysis; the resulting beta-alanol is allowed to react with vanillin (Duquenois reagent) in presence of McIlvain buffer pH 7.5. The color developed is measured at 406 nm. The linearity range was found to be 50-500 microg/ml while the lower limit of detection was about 10 microg/ml. The second is a sensitive and reliable modified fluorimetric method is also suggested, whereas panthenol, after alkaline hydrolysis is treated with ninhydrin. The fluorescent product was found to have excitation lambda(max) at 385 nm and emission lambda(max) at 465 nm. The method showed high sensitivity with linearity range from 0.01 to 3 microg/ml. The lower limit of detection (LOQ) reached 0.005 microg/ml. Validation of both methods was carried out and the two methods were applied for determination of panthenol in some cosmetic and pharmaceutical formulations.