Validation.

This present study was undertaken with an objective to develop & validate a simple, precise, cost effective, sensitive & fast RP- HPLC method for the analysis of Ranolazine. A Shimazu HPLC system with Luna 5µm C18 is employed for the analysis using Methanol: Acetonitrile(50:50, v/v) as mobile phase. Signal from Ranolazine is detected at 227nm by UV Spectrophotometer. The total retention time was 5 min with a flow rate of 1.0 ml/min. % 0f RSD values for precision is found to be 0.798%.The limits of detection (LOD) and quantification (LOQ) were 0.616 and 1.86, respectively. As per ICH guidelines the proposed method is fully validated and found to be linear over a workable drug concentration, accurate, precise and robust. This fast, simple and inexpensive method is suitable for research laboratories & also for quality control analysis in pharmaceutical industries.

A simple and precise stability-indicating high performance liquid chromatography method was developed and validated for the simultaneous estimation of Tolperisone Hydrochloride and Diclofenac Sodium in commercial tablet dosage form. Separation was achieved by using C18 (250mm × 4.6 mm, 5µm) column and mobile phase comprising of potassium dihydrogen phosphate buffer (pH: 6.0): acetonitrile (70:30% v/v) at a flow rate of 1.0ml/min. The detection wavelength was 275 nm. The retention times for Tolperisone Hydrochloride and Diclofenac Sodium were 5.26min and 3.59min, respectively. Linearity of Tolperisone Hydrochloride and Diclofenac Sodium were found in range of 15-52.5µg/ml and 5-17.5µg/ml. The % recovery of Tolperisone Hydrochloride was found to be 99.58%-100.88% and 100.75%-101.33% for Diclofenac Sodium. The values of Limit of detection and limit of quantification were found to be 0.203µg/ml and 0.616µg/ml for Tolperisone Hydrochloride and 0.003µg/ml and 0.010µg/ml for Diclofenac Sodium, respectively. The linear regression coefficient for both drugs was found to be 0.999. The method was found to be robust since the retention times and areas were within the limits even after little deliberate variations in pH, flow rate, mobile phase ratio. Stress studies were conducted on the drug substance and product under the ICH prescribed stress condition viz. hydrolysis, oxidation, photolysis and thermal stress. The drugs showed sufficient decomposition under acidic hydrolysis, alkaline hydrolysis, and oxidation. The drug was found to be moderately sensitive to thermal studies and sunlight studies. This method can be successfully employed for simultaneous quantitative analysis of Tolperisone Hydrochloride and Diclofenac Sodium in pharmaceutical dosage form.

Levocetirizine Dihydrochloride is an orally active, non-sedative antihistamine drug. To determine the assay content of Levocetirizine Dihydrochloride drug substance, a very simple, accurate, specific and precise UV- spectrophotometric method has been built up as well as evaluated. The suggested method comprises dissolving Levocetirizine Dihydrochloride in 0.1M Hydrochloric acid solution and subjecting the consequential solution to UV Spectroscopic measurement. An Absorption maximum was found to lie at about 231nm and the measurements were carried out at this wavelength. Beer's law was followed in the concentration range of 7.5 to 22.5 μg/mL. The linearity showed on the calibration curve between concentration and absorbance by the line equation of y = 0.0377x - 0.0043 (R² = 0.9992). Reproducibility by repeating methods as %RSD was found to be less than 2%. The results of the accuracy and precision were found very satisfactory and here the suggested method was statically validated as per the ICH guidelines in terms of the specificity, linearity, accuracy, precision and robustness. Validation studies have discovered that the method is simple, specific, rapid, reproducible, precise, accurate and economical which is useful for the routine analysis of Levocetirizine Dihydrochloride

The aim of present investigation was to develop a simple UV-visible spectrophotometric method for the determination of Ibuprofen (IBF) in its pure form and marketed formulations and to validate the developed method. Ibuprofen was estimated at UV maxima of 222.8 nm in pH 7.2 phosphate buffer using UV-Visible double beam spectrophotometer. Following the guidelines of International Conference on Harmonization (ICH), the analytical parameters like linearity, precision, and accuracy were studied. The obtained results of analysis were validated statistically and by performing recovery studies to confirm the accuracy of the proposed method. In the developed method, linearity over the concentration range of 2-20 µg/ml of IBF was observed and was found in agreement of Beer’s law. The linear regression was found to be 0.999. The precision (intra-day & inter-day) of method was found within limits (RSD < 2%). The sensitivity of the method was assessed by determining limit of detection and limit of quantification. It could be concluded from the results obtained that the method for estimation of IBF in pure form and in marketed tablets is simple, rapid, accurate, precise and economical and can be used, successfully, in the quality control of pharmaceutical formulations and routine laboratory analysis.

A simple, economic, rapid, high range and accurate stability indicating RP-HPLC method was development for simultaneous estimation of Azithromycin and Ambroxol Hydrochloride in their combined tablet dosage form. This method was carried out by using Isocratic peak HPLC instrument with kromasil C-18 Column (250 mm X 4.6mm,5um) with mobile phase consisting a mixture of Methanol: Acetonitrile: Phosphate buffer in the ratio of 70:20:10 (v/v), at a flow rate of 1.1 ml/min with UV detection at 221nm . The retention time for Azithromycin and Ambroxol Hydrochloride are 9.08 and 5.39min respectively. Suitability, specificity, linearity, accuracy, precision, stability, and sensitivity of this method for the quantitative determination of the drugs were proved by validation in accordance with the requirements laid down by International Conference on Harmonization (ICH) Q2 (R1) guidelines. To establish stability indicating nature of the LC method, forced degradation of drug substances was performed under stress conditions like thermal, oxidation, peroxide, UV light, acid and base hydrolysis) The limit of quantification (L.O.Q) for Azithromycin and Ambroxol Hydrochloride are found to be 0.70ug/ml 1.00ug/ml.Then the limit of detection (L.O.D) for Azithromycin and Ambroxol Hydrochloride are found to be 0.15ug/ml and 0.3ug/ml respectively. The results of the study showed that the proposed method is reliable and robust and can be used as quality control tool for the estimation of these drugs in combined pharmaceutical solid dosage forms.

Two novel methods having requisite precision, accuracy, specificity and robustness were developed and validated for quantitative determination of Apixaban in pharmaceutical dosage forms. The first method was based on isocratic reverse phase liquid chromatography using Sunfire C18, 150mm×4.6mm, 5µ and mobile phase consists of Buffer: acetonitrile (60:40) at a flow rate 1ml/min and detection was achieved photodiodide array detector set at 280 nm. The response was linear range of 5-50 µg/ml (R2 =0.9998). The second spectrophotometric method involves detection at 280 nm. The calibration curve range between 5-50 µg/ml (R2 =0.9999). Validation of method was carried out fulfilling ICH guidelines. Both the methods were applied without any interference from excipients, for determination of drug in coated tablets. It is suggested that the proposed HPLC and UV spectrophotometric methods could be used routine quality control and dosage form assay of Apixaban.

A simple, selective and rapid reverse phase high performance liquid chromatographic (RP-HPLC) method for the analysis of Gliclazide in bulk and in tablet dosage form has been developed and validated. Sample was analysed on a Enable C18 (250mm X 4.6 mm i.d, particle size 5μm) column. The mobile phase consist of Methanol: Water (pH 3.5) in the ratio of 85:15v/v which was sonicated to degased and delivered at a flow rate of 1ml/min at ambient temperature. The retention time of Gliclazidewas 3.7+0.02 minutes. Studies were performed using an HPLC system equipped with a UV detector; the response was monitored at 230 nm.The calibration curve was linear over the concentration range of 20-70 μg/ml (r2=0.999). The limit of detection for Gliclazide was found to be 0.2438 μg/ml and the limit of quantification limit was about 0.7388 μg/ml. The accuracy of the method was established based on the recovery studies. The proposed method can be applied to the routine analysis of Gliclazide in bulk and in tablet dosage form.

A simple, accurate and precise UV spectrophotometric method has been developed for the quantitative estimation of Efavirenz in bulk and tablet dosage form. The λmax was found to be 239 nm. Beer’s law was obeyed in the concentration range of 10-20μg/ml. The regression equation was Y=0.052x-0.113 with value of R2 as 0.996. The method showed good linearity, accuracy and reproducibility. Accuracy as expressed mean percent recovery ± standard deviation was found to be 95.29% ± 0.0429. Percent relative standard deviation values for the intra-day and inter-day precision studies were found to be 0.23 and 0.31, respectively. The limit of detection and limit of quantitation values were found to be 1.39 and 4.22, respectively. Assay of Efavirenz in tablet formulation was performed and percent purity of tablet was found to be 98.65% ± 0.0078.

This research is directed towards Synthesis and quantitation of process-related impurity in Felodipine bulk and formulation. The synthesis of 1,4-Dihydro-2,6-Dimethyl-4-(m- chloro phenyl) pyridine-3,5 Dicarboxylate was identified, characterized, developed and validate by using various analytical techniques such as UV, IR, NMR  for the assessment of impurities in the bulk and formulation in Felodipine. The synthesis of of 1,4-Dihydro-2,6-Dimethyl-4-(m- chloro phenyl) pyridine-3,5 Dicarboxylate was performed by Hantzch pyridine synthesis, by using m-chlorobenzaldehyde, ethylacetoacetate, in presence of ammonia and methanol as a catalyst. The percentage yield was observed to be 80.29%. The preliminary evaluation was performed via melting point, elemental analysis and thin layer chromatography (TLC). Melting point of obtained synthesised compound was noticed to be 134-1370C, whereas Rate of flow ( Rf) value was estimated and found to be 0.70. The TLC of impurity was performed by using Benzene and Methanol (6:1). The structure confirmation of obtained synthesized impurity by using sophisticated analytical instrument viz, Fourier transform infra red( FT-IR), nuclear magnetic resonance (NMR) and ultra violet (UV) spectroscopy. The method was validated as per ICH guidelines and was found to be linear, precise, robust, accurate, rugged.

The study describes the simple, sensitive, accurate, rapid and reliable ultra violet spectroscopic method has been developed for determination of Deferiprone in bulk drug and pharmaceutical formulation. Deferiprone is use as second line agent for thalassemia when iron overload from blood transfusion occurs.in order to investigate the stability of drug, a stress testing of drug sample by exposing it to variety of force degradation conditions has been recommended. Deferiprone was subjected to stress degradation under different condition recommended by international conference on harmonization (ICH). Deferiprone shows maximum absorbance at 279nm & calibration graph linear in the concentration range 5-25 Mcg/ml with correlation co-efficient 0.9997.The higher percentage of recovery study indicates that there is no interference of excipients in the presence of formulation. The stability study indicates appreciable changes were observed by treating the drug with acidic hydrolysis, basic hydrolysis and oxidation. However, there is no appreciable changes were observed for thermal stress and photolytic degradation.