Development Of Spectrophotometric Method Research Articles

Spectrophotometric method development and validation for atorvastatin calcium and nifedipine HCl in bulk and tablet dosage form using absorption ratio method assay of atorvastatin and nifedipine

A simple, economic, accurate Absorption ratio method was developed for the simultaneous estimation of atorvastatin calcium and nifedipine HCl in bulk and tablet dosage form. Methanol was used as a diluent. The absorptions were observed at 237 nm and 297 nm which were selected based on overlap spectra of atorvastatin calcium and nifedipine HCl. The linearity range was found to be 6–10 μg/ml. The proposed method was validated. The reports was expressed that the proposed method was found to be simple, precise, accurate and rapid for the simultaneous estimation of atorvastatin calcium and nifedipine HCl in bulk and tablet dosage form using absorption ratio method.

Development and Validation of Indirect Visible Spectrophotometric Method for Doxepin and Dothiepinin Pure and the Tablet Dosage Forms

The aim of the present work is to develop a simple, sensitive, accurate and rapid visible spectrophotometric method for the quantitative estimation of doxepin and dothiepin in both pure and tablet dosage forms. The developed method is based on the oxidation of DOX and DOT by Ce(IV)inan acid medium followed by reacting it with leuco crystal violetto form a violetcoloreddye which showed absorption maxima (λmax) at 590 nm. Beer's law was obeyed in the concentration range of 0-60μg mL-1in both the drugs. The recovery ranged from 99.26- 100.02% for DOX and from 99.64 – 100.36% for DOT. The regression coefficient was found to be 0.998 and 0.996for DOX and DOT,respectively.The limits of detection (LOD) and quantification (LOQ) values are calculated according to ICH guidelines. The method developed was successfully applied for the determination of DOX and DOT in dosage forms.

Development and validation of ratio-derivative spectrophotometric method for simultaneous estimation of Gabapentin, Methylcobalamin and alpha lipoic acid in tablet formulation

AimThe purpose of this study is to develop simple, economic, specific, rapid, reliable, and reproducible method for simultaneous estimation of the Gabapentin, Methylcobalamin and Alpha lipoic acid. MethodIn this method the overlapping spectra of Gabapentin, Methylcobalamin and Alpha lipoic acid were well resolved by making use of the first-derivative of the ratios of their direct absorption spectra. The derivative ratio absorbance of Gabapentin, Methylcobalamin and Alpha lipoic acid were measured at 731.10 nm, 768.53 nm and 242.21 nm for their quantification. The method was validated for accuracy, precision, linearity, robustness and sensitivity. Result & discussionGabapentin, Methylcobalamin and Alpha lipoic acid were shown linearity in the concentration range of 100–500 μg/ml, 0.5–2.5 μg/ml and 100–500 μg/ml respectively. The LOD & LOQ were found to be 3.09 μg/ml and 9.37 μg/ml; 0.03 μg/ml and 0.10 μg/ml; and 4.79 μg/ml and 14.52 μg/ml respectively. The % labelled claim for Gabapentin, Methylcobalamin and Alpha lipoic acid were found to be 98.71, 98.94 and 98.44 respectively. ConclusionThus, the described method is suitable for routine analysis and quality control of pharmaceutical preparations containing these drugs in combination.

Development and validation of new spectrophotometric ratio H-point standard addition method and application to gastrointestinal acting drugs mixtures

New, simple, specific, accurate and precise spectrophotometric technique utilizing ratio spectra is developed for simultaneous determination of two different binary mixtures. The developed ratio H-point standard addition method (RHPSAM) was managed successfully to resolve the spectral overlap in itopride hydrochloride (ITO) and pantoprazole sodium (PAN) binary mixture, as well as, mosapride citrate (MOS) and PAN binary mixture. The theoretical background and advantages of the newly proposed method are presented. The calibration curves are linear over the concentration range of 5–60μg/mL, 5–40μg/mL and 4–24μg/mL for ITO, MOS and PAN, respectively. Specificity of the method was investigated and relative standard deviations were less than 1.5. The accuracy, precision and repeatability were also investigated for the proposed method according to ICH guidelines.

Development and validation of sensitive spectrophotometric method for determination of two antiepileptics in pharmaceutical formulations

Rapid, sensitive and validated spectrophotometric methods for the determination of two antiepileptics (gabapentin (GAB) and pregabalin (PRG)) in pure forms and in pharmaceutical formulations was developed. The method is based on the formation of charge transfer complex between drug and the chromogenic reagents quinalizarin (Quinz) and alizarin red S (ARS) producing charge transfer complexes in methanolic medium which showed an absorption maximum at 571 and 528nm for GAB and 572 and 538nm for PRG using Quinz and ARS, respectively. The optimization of the reaction conditions such as the type of solvent, reagent concentration and reaction time were investigated. Beer’s law is obeyed in the concentration ranges 0.4–8.0 and 0.5–10μgmL−1 for GAB and PRG using Quinz and ARS, respectively. The molar absorptivity, Sandell sensitivity, detection and quantification limits are also calculated. The correlation coefficients were ⩾0.9992 with a relative standard deviation (RSD%) of ⩽1.76. The methods are successfully applied to the determination of GAB and PRG in pharmaceutical formulations and the validity assesses by applying the standard addition technique, which compared with those obtained using the reported methods.

Use of response surface methodology for development of new microwell-based spectrophotometric method for determination of atrovastatin calcium in tablets

BackgroundResponse surface methodology by Box–Behnken design employing the multivariate approach enables substantial improvement in the method development using fewer experiments, without wastage of large volumes of organic solvents, which leads to high analysis cost. This methodology has not been employed for development of a method for analysis of atorvastatin calcium (ATR-Ca).ResultsThe present research study describes the use of in optimization and validation of a new microwell-based UV-Visible spectrophotometric method of for determination of ATR-Ca in its tablets. By the use of quadratic regression analysis, equations were developed to describe the behavior of the response as simultaneous functions of the selected independent variables. Accordingly, the optimum conditions were determined which included concentration of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), time of reaction and temperature. The absorbance of the colored-CT complex was measured at 460 nm by microwell-plate absorbance reader. The method was validated, in accordance with ICH guidelines for accuracy, precision, selectivity and linearity (r² = 0.9993) over the concentration range of 20–200 μg/ml. The assay was successfully applied to the analysis of ATR-Ca in its pharmaceutical dosage forms with good accuracy and precision.ConclusionThe assay described herein has great practical value in the routine analysis of ATR-Ca in quality control laboratories, as it has high throughput property, consumes minimum volume of organic solvent thus it offers the reduction in the exposures of the analysts to the toxic effects of organic solvents, environmentally friendly "Green" approach) and reduction in the analysis cost by 50-fold.

Development and validation of spectrophotometric method for simultaneous estimation of paracetamol and lornoxicam in different dissolution media

Background: Paracetamol and lornoxicam in combined tablet dosage form are available in the market. This combination is used to treat inflammatory diseases of the joints, osteoarthritis and sciatica. Spectrophotometric and high performance liquid chromatography (HPLC) methods have been reported for their simultaneous estimation in tablet dosage form in specific solvent. This paper presents simple, accurate and reproducible spectrophotometric method for simultaneous determination of paracetamol and lornoxicam in tablet dosage form in different dissolution media. The reported method is helpful in determination of paracetamol and lornoxicam during dissolution study. Materials and Methods: Simple, sensitive, accurate and economical spectrophotometric method based on an absorption correction equation was developed for the estimation of paracetamol and lornoxicam simultaneously in tablet dosage form in different dissolution media at different pH. Results: Paracetamol showed absorption maxima at 243 nm in 0.1N HCland phosphate buffer pH 6.8, while lornoxicam showed absorption maxima at 374 nm in 0.1N HCland phosphate buffer pH 6.8. The linearity was obtained in the concentration range of 4-12 μg/ml for paracetamol and 4-16 μg/ml for lornoxicam. Discussion: The concentrations of the drugs were determined by an absorption correction equation method. The results of analysis have been validated statistically by recovery studies.

Development and validation of Spectrophotometric method and TLC Densitometric Determination of Irinotecan HCl in pharmaceutical dosage forms

The study is focused to develop and validate a UV-Spectrophotometric and Densitometry method for simultaneous estimation of Irinotecan from their dosage form. Based on spectrophotometric characteristic of method are described for the simultaneous determination of Irinotecan HCl, at 247nm for simple UV spectrum and at 268nm for derivative spectrum was found adequate for quantification. The method was validated for linearity, accuracy, precision, reproducibility, and specificity as per International Conference on Harmonization guidelines. The linearity signal and concentration of Irinotecan in the range of 2–10μg/ml in aqueous solution present a correlation coefficient (r2) of 0.9999 for simple UV and 0.9997 for first order derivative spectrum. Second method is the high performance thin layer chromatographic (HPTLC) separation followed by densitometric measurements on normal phase silica gel 60F254. The chromatographic separation was carried out on precoated silica gel 60F254 aluminium plates using a mixture of toluene/ethyl acetate/methanol/carbon tetrachloride, in the volume ratio of 9.2:5:0.9:0.8 (v/v/v/v) respectively as mobile phase. Densitometric analysis was carried out at 317nm. The linear regression analysis data showed good linear relationship in the concentration range of 200–1200 (ng/band) for Irinotecan. The limits of quantitation for Irinotecan were 34 (ng/band). The limit of detection (LOD) and limit of quantitation (LOQ) were 36 and 57ng/spot, respectively. The drug was satisfactorily resolved with Rf value 0.34±0.08. The accuracy and repeatability of the proposed method were ascertained by evaluating various validation parameters like linearity (200–1200ng/spot). The method was found to be rapid, specific, precise and accurate and can be successfully applied for the routine analysis of Irinotecan bulk and marketed dosage form.

Development of spectrophotometric method for simultaneous estimation of diclofenac sodium and papaverine hydrochloride in tablets based on simultaneous equation method

A simple, rapid, accurate and precise spectrophotometric method for simultaneous estimation of diclofenac sodium and papaverine hydrochloride in combined tablet and the dissolution media used in the release studies has been developed. It employs formation and solving of simultaneous equation using two analytical wavelengths corresponding to diclofenac sodium and papaverine hydrochloride in the dissolution medium. This method obeys Beer’s Law in the employed concentration ranges of 2.5-25 µg/mL for two active substances. The method was validated.

Development of difference spectrophotometric method for the estimation of leflunomide in tablet dosage form

A new simple, accurate, precise, highly sensitive and reproducible difference spectrophotometric method for the determination of leflunomide in bulk and pharmaceutical dosage form is described. Difference spectroscopic method is based on the principle that leflunomide exhibit two different forms; in acidic and basic medium which differs in their absorption spectra. The difference spectra were obtained by reading the absorbance of leflunomide in 0.1N HCl in the reference cell and the absorbance of leflunomide in 0.1N NaOH in the sample cell and vice versa; in the difference spectrum maxima and minima were seen at 293.5nm and at 261.5nm respectively. The amplitude values were calculated, which was plotted against concentration. The method is found to be linear in the concentration range of 2-12 ?g/ml. The percentage recovery was found to be between the ranges from 98.92 % to 99.08 %. The proposed method was statistically validated and successfully applied for analysis of tablet dosage forms.

Exploring the Application of Hydrotropic Solubilization Phenomenon for Estimating Diacerein in Capsule Dosage Form by Spectrophotometry Methods

ObjectiveTo develop two safe, novel, eco-friendly, sensitive and accurate UV – spectrophotometric methods by applying hydrotropic solubilization phenomenon for the estimation of diacerein in capsule dosage form. MethodPreliminary solubility studies of drug, selection of hydrotrope, UV spectral studies, Optimization of hydrotrope, direct spectrophotometric and derivative method development, validation of proposed methods were performed as per ICH guidelines. Application of developed method on marketed formulation. ResultsThe aqueous solubility of diacerein was increased by more than 270 folds by using 8 M urea solution as hydrotropic agent in comparison to solubility in distilled water. The sample obeys the Beer's law in the concentration range of 1 – 15 μg/mL & 2 – 45 μg/mL with correlation coefficient of 0.9994 & 0.9997 for each method respectively. The accuracy was proved by recovery studies with mean recovery of 99.80% and 99.08% for each method respectively. Intermediate & repeatability precisions were performed on two consecutive days and analyst to analyst variation with %RSD obtained less than 2%. The LOD & LOQ results intricate the sensitivity of both the methods. ConclusionsIt can be concluded that by applying the hydrotropic solubilization technique for estimating hydrophobic drugs provides a simple, sensitive, cheap & safe estimation. Moreover detrimental health effects & hazardous effects on our environment by using organic solvents can be overcome. Proposed method is less time consuming with two steps of analysis for estimating drug content in formulation.

Studies on the development of spectrophotometric method for the determination of haloperidol in pharmaceutical preparations

A spectrophotometric method based on the formation of ion-pair complex between haloperidol and eriochrome black T (EBT) at pH 1.85 has been described. The formed complex was extracted quantitatively into chloroform and measured at 510 nm. Infra red (IR) studies were performed to confirm the formation of ion-pair complex. Beer’s law was obeyed in the concentration range of 2.0-9.0 μg mL-1 with molar absorptivity of 2.67 × 104 L mol-1 cm-1. The detection limit was found to be 0.18 μg mL-1. Statistical comparison of the results of the proposed method with those of the reference method shows excellent agreement and indicates no significant difference in accuracy and precision.

Development and Validation of Spectrophotometric and HPLC Method for the Simultaneous Estimation of Salbutamol Sulphate and Prednisolone in Tablet Dosage Form

Simple, accurate, and reproducible UV spectrophotometric and HPLC method for simultaneous estimation of salbutamol (SAL) and prednisolone (PRE) was developed in the present work. The first developed method was Simultaneous equation method, wavelength selected are 227 nm for salbutamol and 244 nm for prednisolone respectively. Linearity was observed in concentration range of 6-20μg/ml for salbutamol as well as for prednisolone. Second developed method was RP-HPLC method using Thermo C18 column (4.6 mm i.d × 250 mm) and acetonitrile: 0.025M potassium dihydrogen orthophosphate buffer (pH adjusted to 3.5 with orthophosphoric acid) in the ratio of 30:70% v/v as mobile phase. For HPLC method, linearity was observed in the concentration range of 20-100μg/ml for salbutamol as well as for prednisolone and drugs was subjected to oxidation, hydrolysis, and heat to apply stress condition for degradation studies. Results of analysis were validated stastically and by recovery studies.

Spectrophotometric method development and validation of itopride hydrochloride in bulk and dosage form.

A simple, precise and economical spectrophotometric method for the estimation of Itopride hydrochloride in pharmaceutical bulk and tablet dosage form was developed and validated. Identification was carried out using a UV-visible double beam spectrophotometer detector with working wavelength at 258nm in 0.1N HCl medium. The method was validated with respect to its specificity, linearity range, accuracy, and precision in analytical media. Itopride hydrochloride show the maximum absorbance (λmax) at 258 nm, Regression analysis showed good correlation in the concentration range 5-100 mcg/ml and 96.80 to 106.84% recovery with relative standard deviation 0.293 to 3.98%. Statistical treatment of data reflects that the proposed method is precise, accurate and easily applicable for determination of Itopride hydrochloride in bulk and pharmaceutical preparation. Keywords: Spectrophotometric; Itopride hydrochloride; Validation.

Development and validation of spectrophotometric method for the determination of DPP-4 inhibitor, sitagliptin, in its pharmaceutical preparations

A simple, sensitive and reproducible spectrophotometric method was developed for the determination of sitagliptin phosphate in bulk and in pharmaceutical formulations. The proposed method is based on condensation of the primary amino group of sitagliptin phosphate with acetyl acetone and formaldehyde producing a yellow colored product, which is measured spectrophotometrically at 430nm. The color was stable for about 1 hour. Beer’s law is obeyed over a concentration range of 5-25 μg/ml. The apparent molar absorptivity and Sandell sensitivity values are 1.067 x 104 Lmol-1cm-1 and 0.0471 μgcm-2 respectively. All the variables were studied to optimize the reaction conditions. No interference was observed in the presence of common pharmaceutical excipients. The validity of the method was tested by analyzing sitagliptin phosphate in its pharmaceutical preparations. Good recoveries were obtained. The developed method was successfully employed for the determination of sitagliptin phosphate in various pharmaceutical preparations.

Development and validation of spectrophotometric method for the determination of DPP-4 inhibitor, sitagliptin, in its pharmaceutical preparations

A simple, sensitive and reproducible spectrophotometric method was developed for the determination of sitagliptin phosphate in bulk and in pharmaceutical formulations. The proposed method is based on condensation of the primary amino group of sitagliptin phosphate with acetyl acetone and formaldehyde producing a yellow colored product, which is measured spectrophotometrically at 430nm. The color was stable for about 1 hour. Beer’s law is obeyed over a concentration range of 5-25 μg/ml. The apparent molar absorptivity and Sandell sensitivity values are 1.067 x 104 Lmol-1cm-1 and 0.0471 μgcm-2 respectively. All the variables were studied to optimize the reaction conditions. No interference was observed in the presence of common pharmaceutical excipients. The validity of the method was tested by analyzing sitagliptin phosphate in its pharmaceutical preparations. Good recoveries were obtained. The developed method was successfully employed for the determination of sitagliptin phosphate in various pharmaceutical preparations.

A first-derivative spectrophotometric method for the estimation of Lopinavir in tablets

The present study aimed at the development and validation of a first-derivative ultraviolet (UV) spectrophotometric method for the estimation of Lopinavir in formulation as well as in bulk drug. The reported methods of analysis include chromatographic methods such as high-performance liquid chromatography using a UV detector. Chromatographic methods are tedious, time consuming and are not suitable for routine analysis. However, UV-based methods are fast, simple and economical. Thus, these methods are preferred for routine analysis. The absorption scan of Lopinavir in the UV region does not exhibit any sharp peak. However, the first-derivative spectrum showed an absorption minima at 220 nm, which was selected as the analytical wavelength. Thus, a first-derivative method was developed and validated for the routine analysis of Lopinavir. The method was found to be simple, sensitive and selective, as evidenced by non-interference from the excipients and ritonavir present in the tablet formulation. The linearity was observed in the range of 5 to 35 μg/ml. The method was found to have a high degree of accuracy and precision, both inter- and intra-day. The limit of detection and limit of quantitation were found to be 0.844 and 2.558 μg/ml, respectively.

Development and validation of spectrophotometric method for estimation of anastrozole bulk and pharmaceutical dosage formulation

A simple, precise and accurate method was developed and validated by using a simple solvent system for anastrozole bulk and tablet dosage form. In the developed method, water and ethanol are used as solvents. The λmax was determined to be 221nm. The procedure was validated as per ICH rules for Accuracy, Precision, Detection limit, Linearity, Reproducibility and Quantitation limit. The linearity concentration range was 40-60μg/mL with the correlation coefficient of 0.9971. The percentage recovery for anastrozole was found to be 98.6 to 100.8%. Limit of detection and limit of quantitation values were found to be 1μg/mL and 3μg/mL. The method has been successfully used to analyze commercial solid dosage containing 1mg of anastrozole with good recoveries and proved to be robust. Keywords: Anastrozole, Spectrophotometer, Method development, Validation

Development of Rapid UV Spectrophotometric Method for the Estimation of Efavirenz in Formulations

Simple, sensitive, accurate, precise and rapid ultraviolet (UV) spectrophotometric method was developed for the estimation of efavirenz in pure form, its formulations and stability samples. For the estimation of efavirenz, solvent system employed was 1% w/v sodium lauryl sulphate (SLS) and wavelength of detection (λdet) was 247 nm. The developed method was used to estimate the total drug content in two commercially available oral formulations of efavirenz and recovery studies were also carried out. Sample recovery in both the formulations using the above method was in good agreement with their respective labeled claims, thus suggesting the validity of the method and non‐interference of formulation excipients in the estimation. The developed method was found to be stability specific and were validated as per ICH guidelines‐2005, USP‐2000 and statistical methods.

Spectrophotometric determination of some amino heterocyclic donors through charge transfer complex formation with chloranilic acid in acetonitrile

The present study is directed toward a development of simple, rapid and accurate spectrophotometric method for determination of some amino heterocyclic donors. The Charge transfer (CT) interactions between 3-aminopyrazole (AP), 3,5-dimethyl-pyrazole (DMP), 3-amino-5-methyl-pyrazole (AMP), 2-amino-4-methyl-thiazole (AMT), 2-amino-5-methyl-1,3,4-thiadiazole (AMTD) and 2-amino-5,6-dimethyl-1,2,4-triazine (ADMT) with chloranilic acid (CLA) as π-acceptor have been investigated spectrophotometrically in acetonitrile. Factors controlling the CT-reactions were also studied and optimized. Under the optimum conditions, linear relationships with good correlation coefficients were identified with high accuracy and precision. Job's method of continuous variation, spectrophotometric and conductometric titrations were used to identify the composition of the formed CT-complexes. Benesi-Hildebrand (BH) equation has been used to calculate the formation constants of the charge transfer reactions K CT and the molar extinction coefficients ε. Free energy change (Δ G o), oscillator strength ( f), transition dipole moment ( µ), ionization potential ( I P) and charge transfer energy of the formed CT-complexes ( E CT) were also determined and evaluated.