ICH Q2 R1 Guidelines Research Articles

Validated green and white RP-HPLC method for the estimation of Zolmitriptan in marketed dosage form

To develop and validate a rapid, accurate, economical, effective and greenery RP-HPLC method for the determination of Zolmitriptan in tablet dosage form. RP-HPLC method was developed using Luna (C18) (4.6×250mm, 5μm) column and Sodium phosphate buffer (pH 4.7): Methanol [75: 25, v/v] was used as mobile phase at a flow rate of 1.0mL/min. The detection was carried out at 227nm. Further, eco-friendliness, productivity and performance of the optimized analytical method were assessed by green and white tools. The retention time of Zolmitriptan was found to be 3.25min with acceptable chromatographic parameters. The optimized RP-HPLC method was more eco-friendly, efficient, throughput and practicable than the reported methods as confirmed by AES, AGREE, GAPI and RGB tools. Further, the proposed analytical method showed all the validation parameters within the acceptance limit of ICH Q2 R1 guidelines. The linear regression analysis indicated a good linear response in the 10 to 120μg/mL concentration range with R2 of 0.99998. The percentage content and percentage assay of Zolmitriptan in Zomig-5mg tablet was found to be 103.36±0.356% and 97.86±0.693%. The developed and validated method has several advantages compared to the reported HPLC methods and is useful in the systematic analysis of Zolmitriptan in its dosage form.

Simultaneous UV Spectrophotometric Method for Estimation of Nirmatrelvir and Ritonavir in Bulk and Tablet Dosage Form

A simple, specific, precise, and accurate UV spectrophotometric method has been created for the simultaneous measurement of Nirmatrelvir and Ritonavir in pharmaceutical dosage forms. The absorption maxima of the drugs were found to be at 240 nm and 258 nm for Nirmatrelvir and Ritonavir respectively. Nirmatrelvir and Ritonavir obeyed Beer’s law in the concentration range of 12-18 µg/ml and 8-12 µg/ml respectively. Different analytical parameters such as linearity, precision, accuracy, limit of detection (LOD) and limit of quantification (LOQ) were determined as per ICH guidelines. Limit of detection and quantification values for Nirmatrelvir 1.25 and 3.85 μg/ml and for Ritonavir 0.34 and 1.12 μg/ml respectively. The accuracy of the methods was assessed by recovery studies and recovery values between prescribed limit of 99-101 % shows that method is free from interference of excipients present in formulation. The developed method was free from interferences due to excipients present in formulation and it can be used for routine quality control analysis. The results were validated statistically as per ICH Q2 R1 guideline and were found to be satisfactory. The proposed methods were successfully applied for the determination of for Nirmatrelvir and Ritonavir in commercial pharmaceutical dosage form

A green micelle-enhanced first derivative synchronous fluorescence approach for determination of donepezil HCl and trazodone HCl in their pure state, pharmaceutical dosage form and spiked human plasma.

The study's objective is to establish an eco-friendly, sensitive and economical quantitative methodology for the concurrent analysis of donepezil HCl (DPZ) and trazodone HCl (TRZ) in raw materials, tablets and human plasma. The first derivative synchronous fluorescence spectroscopic (FDSFS) technique was applied at constant wavelength difference (∆λ = 120) for assessment of DPZ and TRZ at each other's zero-crossing point at 279 nm and 297 nm, respectively. The submitted technique was validated in accordance with ICH Q2 R1 guidelines and the linearity of the standard calibration curve was observed over the concentration range of 10-500 ng/ml for DPZ and 20-1,000 ng/ml for TRZ. The detection limits (LOD) were found to be 2.65 and 5.4ng/ml, and the limits of quantitation (LOQ) were 8.05 and 16.3ng/ml for DPZ and TRZ, respectively. This technique was used further to quantify the studied medications in their laboratory-prepared mixtures, commercial tablets and spiked plasma samples. The results obtained were not significantly different from those acquired from the comparison methods, indicating the high accuracy and precision of the proposed method. Furthermore, the ecological friendliness of the suggested method was evaluated and proven to be excellent using Green Analytical Procedure Index (GAPI) and Analytical GREEnness (AGREE) evaluation tools.

Novel HPLC Method Validation for Quantification of Dapagliflozin in Bulk Drug and Formulation

A new, steady, easy-to-use, and straightforward high-performance liquid chromatography (HPCL) technique was also tested to determine dapagliflozin concentration in medicinal and API forms. For the chromatographic extraction study, a Phenomenex Gemini-NX C18 Column was used. This column was filled with a mobile phase that was 70:30 v/v methanol to sodium 1-octanesulphonate. This experiment used a 20-liter syringe amount and a flow rate of 1-mL per minute. A bunch of factors were used to make sure the new method worked. The drug dapagliflozin was released at a wavelength of 203 nm after 6.5 ± 0.5 minutes. A correlation coefficient (r2) of 0.9994 between 15 to 55 μg/mL shows that the planned process is linear. The theoretical plates and peak tailing were found to be 4129 and 1.03, respectively. In terms of healing, the percent accuracy that was reached was within the acceptable range of 98 to 100. New suggestions say that the regular testing of dapagliflozin in large amounts must be done using a proven method that is unique and easy to repeat. Forced breakdown tests with drugs were used to show that the proposed method is stable. The product was tested according to the steps laid out in the ICH Q2R1 Guidelines. The test looked at many chemical processes, such as oxidation, photolysis, heat stress, and acids and alkaline hydrolysis.

A green perspective on simultaneous HPLC and UV spectrophotometric estimation of Udenafil and Dapoxetine hydrochloride in pharmaceutical formulations

Clinical research has affirmed that the combination of Udenafil and Dapoxetine hydrochloride yields greater improvement in erectile dysfunction compared to the use of individual medications. There is a requirement of precise quantitative method for accurate determination of Udenafil and Dapoxetine when taken together. Consequently, our interest lies in developing and validating analytical methods for the simultaneous estimation of Udenafil and Dapoxetine - Various strategies, such as UV spectrophotometric approaches and an innovative, straightforward, accurate, sensitive, and robust High-Performance Liquid Chromatography (HPLC) procedure, have been explored. An Isocratic HPLC approach was employed using reversed-phase CHEMSIL ODS C18 column (4.6 mm x 250 mm, 5 µm) as a stationary phase and detection wavelength (λ max) 230 nm for the two analytes in the chromatographic column. The mobile phase comprised of (70:30, v/v) mixture of Acetonitrile and 10 mM Ammonium formate buffer having pH 5, adjusted with formic acid. The elution was carried out at a constant flow rate of 1 mL/min. The UV simultaneous equation technique estimated the medicine's quantities and derived a simultaneous equation for DAP and UDN using 2 wavelengths, 230 nm and 298 nm. Furthermore, the two advanced methods underwent rigorous statistical validation, guided by the valuable ICH Q2R1 guidelines, affirming their effectiveness as a successful analytical tool for the concurrent analysis of both tablets in bulk and the synthetic mixture fabricated in the laboratory. To assess the environmental impact of contemporary UV spectrophotometric and HPLC techniques, the Green Analytical Greenness Metric (AGREE) software and the Green Analytical System Index (GAPI) were employed to evaluate the greenness profile. This examination confirmed the method's environmental friendliness concerning solvent usage, chemical ingredients, energy consumption, and waste production. The developed UV method is more sustainable and greener compare to HPLC method for simultaneous estimation of compound, which can be utilised by industry for quantification of each individual drug in their combination products.

ANALYTICAL UV SPECTROSCOPY METHOD DEVELOPMENT AND VALIDATION STUDIES FOR SIMULTANEOUS ESTIMATION OF METFORMIN HCL AND QUERCETIN

In the current investigation, we have designed and assessed a simple and swift analytical approach employing UV spectroscopy for the simultaneous quantification of the analytes metformin and quercetin with excellent precision and accuracy. The wavelengths of interest are the wavelengths at which both the drugs show maximum absorbance: 233 nm for metformin and 256 nm for quercetin. Linearity study, conducted in methanol and phosphate buffer, yielded a correlation coefficient (r2 ) of 0.99. The validation study for the developed method was conducted in accordance with ICH Q2 R1 guidelines. The percent recovery was 95% to 105%, and the percent relative standard deviation was <2, demonstrating the accuracy and precision of this method. This method can be applied to analysis of the two compounds in fixed dose formulations using simple UV spectroscopy.

Stability Indicating RP-HPLC-DAD method for simultaneous estimation of tadalafil and macitentan in synthetic mixture for treatment of pulmonary arterial hypertension

High Performance liquid chromatography is an integral analytical tool in assessing drug product stability. A simple, selective, precise, accurate and stability indicating RP-HPLC method was developed and validated for analysis of Tadalafil and Macitentan in synthetic mixture. Chromatographic separation was performed using Phenomex Gemini C18 (25cm×4.6nm, 5μm) Column. The mobile phase consists of (10mM Ammonium Acetate in water and [Methanol: ACN 20: 80% v/v]) (40: 60% v/v). The flow rate was set to be 1.0mL/min. The injection volume was 10.00μL. The detection was carried out at 260nm at column temperature 35°C. The method was validating according to ICH Q2R1 guideline for accuracy, precision, reproducibility, specificity, robustness and detection and quantification limits. Stability testing was performed on Tadalafil and Macitentan and it was found that these degraded sufficiently in all applied chemical and physical conditions. Linearity for Tadalafil and Macitentan was observed 0.4-100μg/mL and 0.1-25μg/mL with correlation coefficient at 0.9999. LOD and LOQ 0.008μg/mL and 0.024μg/mL and 0.001μg/mL and 0.0029μg/mL for Tadalafil and Macitentan respectively. The developed RP-HPLC method was found to be suitable for the determination of both the drugs.

Development and Validation of a Stability-indicating UPLC-DAD Method for the Simultaneous Determination of Ivermectin and Praziquantel in Pharmaceutical Tablets and Dissolution Media.

Currently, there is no single rapid and accurate stability-indicating quantitative method that can simultaneously determine both ivermectin and praziquantel and their related compounds. Thus, the goal of this research is to develop and validate a new rapid, accurate, and stability-indicating ultra-performance liquid chromatography (UPLC) method. The method uses a water, acetonitrile, and methanol gradient. The chromatographic separation was achieved on a C18 (1.7 μm, 2.1 × 50 mm) column with a flow rate of 0.7 mL/min, and the column temperature was maintained at 40°C. Analytes are detected at 245 nm. The method was validated in accordance with ICH Q2R1 guidelines. The linearity (R2) was >0.9987 and 0.9997 for praziquantel and ivermectin, respectively. The corresponding accuracy ranged between 98.0 and 102.0%. Intermediate precision (assessed as inter-day precision) was determined by calculating the cumulative %CV of eighteen assay preparations and was less than 2.0% for both praziquantel and ivermectin. The specificity of the method was shown by the resolution of the two active pharmaceutical ingredients (APIs) from any interfering excipients, impurities, or degradation products. The limit of detection and quantitation for ivermectin was 26.80 ng/mL and 81.22 ng/mL, respectively. The limit of detection and quantitation for praziquantel was 1.39 μg/mL and 4.22 μg/mL, respectively. The robustness study proved that method performance is stable against small variations in sample processing parameters (shaking, sonication time, and acetonitrile % in solvent solution) and also against small variations in the initial % of mobile phase components and gradient slope. Using ICH Q2R2 criteria, the method was demonstrated to be specific, accurate, stability indicating, and robust to small variations of chromatographic variables.

Development and validation of stability indicating RP-HPLC method for determination of aspirin and pantoprazole sodium in synthetic mixture stability indicating HPLC method

The present study describes a new accurate and precise stability indicating reverse phase HPLC method for quantitative computation of pantoprazole sodium and Aspirin from physical simulated mixture. The proposed chromatographic method employs Hypersil ODS C18 column (250 x 4.6 mm, 5μ) as the stationary phase and combination of methyl alcohol and water in ratio of 70: 30 v/v as the elution medium. Overall separation was carried out at 0.8 ml/minute flow rate and elution was monitored at 254 nm. The proposed system gave well resolved peak of Aspirin and Pantoprazole sodium with elution time of 2.32 and 5.85 minute respectively. Same system was effective in separation of active components and degradation products when the components were subjected to forced degradation as per regulatory guidelines (ICHQ1). Finally, the optimized method was successfully validated as per ICH Q2R1 guidelines and applied for quantitative analysis of both active components in synthetic mixture.

Development and validation of spectroscopic simultaneous equation method for simultaneous estimation of Azelnidipine and Valsartan in synthetic mixture

A simple, specific, accurate and precise spectrophotometric method was developed and validated for simultaneous estimation of Azelnidipine and Valsartan in Synthetic Mixture. The wavelength of estimation for Azelnidipine was 240.00 nm and for Valsartan was 250.00 nm. Beer’s law is obeyed in the concentration range of 2-10μg/ml and 16-80μg/ml and correlation coefficient of 0.9999 and 0.9993 for Azelnidipine and Valsartan respectively. The % recovery for Azelnidipine and Valsartan were found to be 101.2 % - 101.9 % and 101.8 - 101.9 % respectively. Intraday precision of Azelnidipine and Valsartan were found to be 0.06 – 0.24 and 0.08 – 0.27 % RSD and Interday precision were found to be 0.14 – 0.33 and 0.11 – 0.37 % RSD respectively. The proposed method was also evaluated by the Assay of Synthetic mixture containing Azelnidipine and Valsartan. The % Assay was found to be 100.50 ± 0.002 for Azelnidipine and 100.87 ± 0.001 for Valsartan. Validation of proposed methods was carried out according to ICH Q2R1Guidelines. The proposed methods were found accurate and reproducible for routine analysis of both the drugs in synthetic mixture.

Development and validation of spectroscopic simultaneous equation method for simultaneous estimation of betamethasone and luliconazole in synthetic mixture

A simple, specific, accurate and precise spectrophotometric method was developed and validated for simultaneous estimation of Betamethasone and Luliconazole in Synthetic Mixture. The wavelength of estimation for Betamethasone was 243.20 nm and for Luliconazole was 225.00 nm. Beer’s law is obeyed in the concentration range of 10-50μg/ml and 20-100μg/ml and correlation coefficient of 0.999 and 0.999 for Betamethasone and Luliconazole respectively. The % recovery for Betamethasone and Luliconazole were found to be 100.49 % & - 100.68 % respectively. Intraday precision of Betamethasone and Luliconazole were found to be 0.10 %– 0.28 % and 0.11 % – 0.24 % RSD and Interday precision were found to be 0.27 %– 0.37 % and 0.23 %– 0.35 % RSD respectively. The proposed method was also evaluated by the Assay of Synthetic mixture containing Betamethasone and Luliconazole. The % Assay was found to be 100.60 % for Luliconazole and 100.52 % for Betamethasone. Validation of proposed methods was carried out according to ICH Q2R1Guidelines. The proposed methods were found accurate and reproducible for routine analysis of both the drugs in synthetic mixture.

Method development and validation of Atorvastatin, Ezetimibe and Fenofibrate using RP-HPLC along with their forced degradation studies and greenness profiling

ABSTRACT The present study aimed to separate the three-drug combination and degradant products by applying stress studies in bulk and pharmaceutical using RP-HPLC green analytical chemistry principles to make the separation better and more effective without compromising the standard limits. The three drugs have separated using the Mobile Phase Mixture of 10 volumes of buffer (0.1% triethanolamine in water) and 90 volumes of ethanol on the stationary phase Inertsil ODS 3 (250 mm x 4.6 mm), 5 µm column, at a wavelength of 256 nm. The method has been performed on a linear concentration range of 12–28 µg/mL for ATR, EZB, and 192- 448 µg/mL for FNF. The forced degradation studies showed that the ATR had > 5% degradation in acid, peroxide and thermal degradation, as EZB tends to degrade more than 15% at alkali hydrolysis. Moreover, the methodology has been validated according to ICH Q2 R1 guidelines, assessed using green evaluation tools like GAPI, AES, and AGREE, and found eco-friendly according to the output of the greenness outcomes. The developed method employed for determining three drugs has shown within limits according to the guidelines, easily adaptable by the pharmaceutical industries for their regular quality control analysis of the combinations.

Development and Validation of UV-Visible spectroscopic method for estimation of Nimesulide in bulk and its pharmaceutical formulation

The aim of the present work was to develop and validate a simple UV Spectroscopic method for the determination of Nimesulide, which is the inhibitor of prostaglandin synthesis from arachidonic acid and platelet aggregation. It has the ability to exert analgesic and anti-inflammatory actions through various mechanisms (ie, COX independent pathways) and also it inhibits the release of tumour necrosis factor (TNFα) and interleukins, and stops the release of histamine from mast cells and reduce the synthesis of platelet activating factor (PAF) from the basophil cells. The UV visible Spectrophotometric analysis was performed using systronics UV-Spectrophotometer 2704 X visible double beam by using solvent KOH. Detection was performed at a wavelength of 395nm. Method validation was carried out according to ICH Q2R1 guidelines by taking the parameters such as accuracy, linearity, precision, ruggedness, and robustness, LOD and LOQ. The UV spectrophotometric was found linear in the range 5- 25µg/ml. The method was rugged and robust with % relative standard deviation less than 2. The extraction recoveries was found to be higher than 99.19% in all experimental conditions. Based upon the performance characteristics, the proposed method was found accurate, precise, rapid and suitable for the determination of nimesulide for routine analysis. The aim of the present work was to develop and validate a simple UV Spectroscopic method for the determination of Nimesulide, which is the inhibitor of prostaglandin synthesis from arachidonic acid and platelet aggregation. It has the ability to exert analgesic and anti-inflammatory actions through various mechanisms (ie, COX independent pathways) and also it inhibits the release of tumour necrosis factor (TNFα) and interleukins, and stops the release of histamine from mast cells and reduce the synthesis of platelet activating factor (PAF) from the basophil cells. The UV visible Spectrophotometric analysis was performed using systronics UV-Spectrophotometer 2704 X visible double beam by using solvent KOH. Detection was performed at a wavelength of 395nm. Method validation was carried out according to ICH Q2R1 guidelines by taking the parameters such as accuracy, linearity, precision, ruggedness, and robustness, LOD and LOQ. The UV spectrophotometric was found linear in the range 5- 25µg/ml. The method was rugged and robust with % relative standard deviation less than 2. The extraction recoveries was found to be higher than 99.19% in all experimental conditions. Based upon the performance characteristics, the proposed method was found accurate, precise, rapid and suitable for the determination of nimesulide for routine analysis.

Development and Validation of Stability-Indicating RP-HPLC Method for the Estimation of Fisetin in Novel Cubosomal Nanoformulation: Application to the Marketed Formulation and Selected Plant Extracts

Background: Fisetin (FIS) is a bioactive flavonoid found in various plants, reported for many pharmacological activities, and presently marketed as a nutraceutical. To overcome less water solubility and bioavailability issues, FIS cubosomal nanoformulation has been prepared and characterized. Objective: To estimate FIS in prepared novel cubosomes, an RP-HPLC analytical method development with the most sensitivity, economical, robust, and wide applicability in marketed FIS formulations and plant extracts also. Method: An RP-HPLC method was developed and validated as per ICH Q2R1 guidelines by using C-18 Phenomenex Luna 5μ, 100A0 column, LC-20 AD pump, and Shimadzu LC solution 1.25 software. The combination of acetonitrile and formic acid (0.1%v/v) in the ratio of 25:75 v/v was used as a mobile phase for chromatographic separation using a PDA detector at 360 nm and a flow rate of 1 ml/min. Results: The developed method was remarkably linear in the range of 0.1 to 16 μg/ml (R2 ˃ 0.999). This method was found to be accurate (recovery 98.24 to 100.65 %), precise, robust (% RSD ˂ 2), and more sensitive than the earlier reported method with LOD and LOQ values of 17.26 and 52.31 ng/ml, respectively. The FIS estimation was also performed using the developed method in the marketed FIS formulation Doctor’s Best ® Fisetin, and different plant extracts such as strawberry, grapes, black tea, and green tea. The forced degradation study suggests that FIS was unstable in alkaline and oxidative stress conditions. Conclusion: For FIS estimation in cubosomal nanoformulation, a widely applicable, novel, robust, most sensitive, and economical RP-HPLC method was developed and validated and also applied to marketed formulations and plant extracts.

Simultaneous UV Spectrophotometric Method for Estimation of Sitagliptin Phosphate and Metformin Hydrochloride in Bulk and Tablet Dosage Form

Two simple, precise and economical UV methods have been developed for the simultaneous estimation of Sitagliptinphosphate and Metformin hydrochloride in bulk and pharmaceutical dosage form. Method A is Absorbance maximamethod,whichisbasedonmeasurementofabsorptionatmaximumwavelengthof266nmand232nmforSitagliptinphosphateandMetforminhydrochloriderespectively.MethodBisareaundercurve(AUC),inthewavelength range of244-279 nm for Sitagliptin phosphateand222-240nm for Metformin hydrochloride. Linearityfordetector response wasobserved in the concentration range of25-225µg/ml for Sitagliptin phosphate and 2-12µg/ml for Metformin hydrochloride. The accuracy of the methods was assessed by recovery studies and was found tobe99.64%and98.98%forSitagliptinphosphateandMetforminhydrochloride.Thedevelopedmethodwasvalidatedwithrespecttolinearity,accuracy(recovery),precisionandspecificity.Theresultswerevalidatedstatistically as per ICH Q2 R1 guideline and were found to be satisfactory. The proposed methods were successfullyappliedforthedeterminationofforSitagliptinphosphateandMetforminhydrochlorideincommercialpharmaceuticaldosage form.

Staility-Indcatng RP-HPLC Method Development and Validation for Simulaneous Estimaton of Gliclazde and Chromium Picolinate in Bulk and Formulaton

Simultaneous determination of Gliclazide and Chromium Picolinate two component methods has been developed in methanol. In this method, the overlapping spectra of Gliclazide and Chromium Picolinate were well resolved by making use of the zero order spectra of their direct absorption spectra. The method was based on the measurement of absorbance of Gliclazide and Chromium Picolinate at 272.00nm, 262.00 nm and 272.05nm respectively. This method obeyed Beer’s law in the concentration range of 0 to 120 μg/mL for Gliclazide and Chromium Picolinate. The method was validated as per the ICH guideline and accuracy, precision are found to be within the acceptable limit. The limits of detection and quantitation were found to be 0.023 and 0.070 μg/ml, respectively for Gliclazide, and 0.021 and 0.063 μg/ml, respectively for Chromium Picolinate . A simple, specific, accurate and precise Simultaneous Equation Method for three Components Spectrophotometric methods was developed and validated for simultaneous estimation of Gliclazide and Chromium Picolinate in synthetic mixture. This method is validated as per ICH Q2R1Guidelines.

Development and validation of an analytical method for the simultaneous estimation of Etoricoxib and Thiocolchicoside in tablet dosage form by UV Spectrophotometric method

A simple, accurate, precise, and rapid UV spectrophotometric method was developed for simultaneous estimation of Etoricoxib and Thiocolchicoside in the tablet dosage form. Etoricoxib showed maximum absorbance at 270 nm, while Thiocolchicoside showed maximum absorbance at 260 nm. These two wavelengths were selected for further analysis. The developed method was validated as ICH guidelines including parameters as linearity, range, precision, LOD, LOQ, and accuracy. The absorbance range for Etoricoxib (5-30 µg/ml) was found to be 0.210-0.799. These aliquots obeyed Beer-Lambert’s law with regression of 0.995. The absorbance range of Thiocolchicoside (5-30 µg/ml) was found to be 0.147-0.468 with a regression of 0.998. The method of precision (interday, intraday) was found within limits (RSD>2). The % recovery was found to be within the range. As per the ICH Q2 R1 guideline, the result was validated statistically and was found to be satisfactory.

Development and validation of UV Spectrophotometric method for simultaneous estimation of Efonidipine hydrochloride ethanolate and Chlorthalidone in their synthetic mixture

Efonidipine Hydrochloride Ethanolate and Chlorthalidone is used in management of hypertension and under clinical phase 3 study. The development of quality control method is required for accurate analysis of both drugs. Two simple, precise and economical UV spectrophotometric methods have been developed for the simultaneous estimation of Efonidipine Hydrochloride Ethanolate and Chlorthalidone in their synthetic mixture. Method I is simultaneous equation method (Vierodt’s Method), which is based on measurement of absorption at 251 and 227 nm i.e. λmax of Efonidipine Hydrochloride Ethanolate and Chlorthalidone, respectively. Method II is first order derivative was based on the measurement of absorbance of Efonidipine Hydrochloride Ethanolate measure at 283.2 nm (ZCP of Chlorthalidone) and absorbance of Chlorthalidone measure at 250.8 nm (ZCP of Efonidipine Hydrochloride Ethanolate). Linearity was observed in the concentration range of 6.4-38.4 µg.mL-1 for Efonidipine hydrochloride ethanolate and 2-12 µg.mL-1 for Chlorthalidone using methanol as a solvent. The accuracy of methods was assessed by recovery studies and was found to be within range of 98-102% for both the drugs. Precision of the methods was estimated by repeatability and intermediate precision studies. The % RSD values were found to be less than 2, proving methods were precise. Two methods were compared using F- test. The results were validated statistically as per ICH Q2 R1 guideline and were found to be satisfactory.

Stability Indicating UV-Spectrophotometric Method Development and its Validation for the Determination of Imatinib Mesylate in Bulk and Formulation

A Simple and precise UV-spectroscopic method was developed for the determination of Imatinib Mesylate in its bulk and formulation. The developed method has been validated for various parameters like specificity, accuracy, linearity, robustness according to USP general chapter<1225> and ICH Q2R1 guidelines. Pure solution of Imatinib Mesylate was scanned in the whole range of UV region where it has shown the maximum absorbance at 258nm. The RSD values for method precision and intermediate precision were found to be well within the acceptance criteria and the series of dilutions were found to be linear (2-12ug/ml) where r2= 0.999 was the regression value. Limit of detection (0.2925μg/ml) and Limit of quantification (0.8977μg/ml) of IMT were established. Further, the drug has been subjected to various stress conditions, and percent degraded was reported. Drug solutions have shown stability on the benchtop for up to 8 hours and 72 hours when the solutions were refrigerated.

A STABILITY INDICATING RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF DAUNORUBICIN AND CYTARABINE IN BULK AND PHARMACEUTICAL DOSAGE FORM

Objective: A simple, accurate, and precise method was developed for the simultaneous estimation of daunorubicin and cytarabine dosage form using reverse-phase HPLC and validated with different parameters such as accuracy, precision, repeatability, linearity, limit of detection (LOD), and limit of quantitation (LOQ) as per ICH Q2R1 guidelines. Methods: The chromatogram was run through Agilent C18 Column of dimensions 150×4.6 mm, 5 m. Mobile phase containing 0.01N KH2PO4: Methanol taken in the ratio 50:50 was pumped through the column at a flow rate of 1.0 ml/min. The temperature was maintained at 30°C. The optimized wavelength selected was 240 nm. Results: Retention times for daunorubicin and cytarabine were found to be 2.433 min and 3.045 min. The %RSD of the daunorubicin and cytarabine was found to be 0.7 and 0.4, respectively. The %Recovery was obtained as 99.96% and 100.40% for daunorubicin and cytarabine, respectively. LOD and LOQ values obtained from regression equations of daunorubicin and cytarabine were 0.08 μg/ml, 0.24 μg/ml and 0.94 μg/ml, 2.86 μg/ml, respectively. The regression equation for daunorubicin is y=28587x+3141 and y=35995x+37534 for cytarabine. Conclusion: The method developed was simple and economical that can be adopted in regular quality control tests in industries.