Validation

  The present work deals with development and validation for simultaneous determination of antihistaminic drugs in pharmaceutical formulations. A rapid, precise and specific high performance liquid chromatography (RP-HPLC) method was developed for Levocetirizine dihydrochloride and Phenylephrine. Chromatographic separations was achieved on Waters Younglin system C-18 (5μm, 250×4.6 mm) HPLC column within a short runtime of 10 min. HPLC system having isocratic mode, with mobile phase containing methanol : water (pH 3) (70:30% v/v) and flow rate maintained at 1.0 mL/min was used. Effluents were monitored at 230 nm. Retention time of Levocetirizine dihydrochloride and Phenylephrine were found to be 2.6 and 4.6 min respectively. Linearity was studied in the concentration range of 2 to 12 μg/mL and 12 to 72 μg/ mL for Levocetirizine dihydrochloride and Phenylephrine respectively, with a correlation coefficient of 0.998 and 0.999 respectively. The proposed method was validated according to the ICH guidelines with respect to specificity, linearity, accuracy, precision and robustness.

A Reversed-Phase High Performance liquid chromatographic (RP-HPLC) method was developed for the simultaneous determination of Ambroxol Hydrochloride and Cefpodoxime Proxetile in combined tablet dosage form. The analysis was carried out using Phenomenex Luna C – 18, pre-packed column. Mobile phase, containing Acetonitrile: 0.05 M Potassium Dihydrogen Ortho Phosphate Buffer (70:30) pH adjusted to 6.7 with Tri ethyl Amine was pumped at a flow rate of 1.0 mL/min with UV-detection at 245 nm. Retention time was 3.34 ± 0.01 min and 4.77 ± 0.01 min for Ambroxol Hydrochloride and Cefpodoxime Proxetile, respectively. The method was validated for linearity, accuracy, precision, and specificity. The method showed good linearity in the range of 30 - 60 μg/ml for Ambroxol Hydrochloride and 50 - 100 μg/ml for Cefpodoxime Proxetile. The detection limit of the proposed method was 4.56 and 12.51 μg/ml and the quantification limit was 13.82 and 37.92 μg/ml for Ambroxol Hydrochloride and Cefpodoxime Proxetile, respectively. The % recovery was within the range between 99.57% and 100.27% for Ambroxol Hydrochloride and % recovery was within the range between 99.89% and 100.86% for Cefpodoxime Proxetile. The % R.S.D for precision and accuracy of the method was found to be less than 2%. The method was validated as per the ICH guidelines. The method was successfully applied for routine analysis of Ambroxol Hydrochloride and Cefpodoxime Proxetile in combined tablet dosage form.

The present research work discusses the two new, simple, accurate, precise and reproducible UV spectrophotometric methods have been developed and validated for the simultaneous determination of Prasugrel (PRASU) and Aspirin (ASP) in their combined dosage form.  Method- I is based on simultaneous equation method using two wavelengths, 254 nm (λmax of PRASU) and 276 nm (λmax of ASP). Method - II Q‐absorption ratio method using two wavelengths, 274.7 nm (Isoabsorptive point) and 254 nm (λmax of PRASU). Methanol was the solvent used in all methods. This method obeyed Beer’s law in the concentration range of 5-60 µg /ml for PRASU and 20-140 μg/ml ASP. All methods were validated statistically and recovery studies were carried out. Hence, the methods herein described can be successfully applied in quality control of combined pharmaceutical dosage form.

The present manuscript describe simple, sensitive, rapid, accurate, precise and cost effective dual wavelength spectrophotometric method for the simultaneous determination of Rifampicin and Piperine in combined capsule dosage form. The utility of dual wavelength data processing program is its ability to calculate unknown concentration of components of interest in a mixture containing an interfering component. The principle for dual wavelength method is “the absorbance difference between two points on the mixture spectra is directly proportional to the concentration of the component of interest”. The method was based on determination of Rifampicin at the absorbance difference between 286 nm and 357 nm and Piperine at the absorbance difference between 356 nm and 479 nm. The linearity was obtained in the concentration range of 10-60 μg/ml for Rifampicin and 1-10 μg/ml for Piperine. The mean recovery was 98.40 ± 0.48 and 98.59 ± 0.46 for Rifampicin and Piperine, respectively. The method was successfully applied to pharmaceutical dosage form because no interference from the capsule excipients was found. The suitability of these methods for the quantitative determination of Rifampicin and Piperine was proved by validation. The proposed methods were found to be simple and sensitive for the routine quality control application of Rifampicin and Piperine in pharmaceutical capsule dosage form. The results of analysis have been validated statistically and by recovery studies.

A simple, rapid and specific RP-HPLC method has been developed and validated for determination of Carvedilol in bulk and tablet formulations. Chromatographic separation was performed by Phenomenex Luna C-18 (250 x 4.6mm, 5μm particle size) column with a mobile phase consisting of a mixture of phosphate buffer, acetonitrile and methanol in the ratio (30:45:25 v/v/v), pH adjusted to 4.8 with ortophosphoric acid. The mobile phase was was filtered through a 0.45μ cellulose nitrate filter, sonicated for 15 min and delivered at a flow rate of 1ml/min. Detection was performed at a wave length of 241 nm at ambient temperature. Linearity was obtained in a concentration range of 30 to130 µg/ml with a correlation coefficient (r2) of 0.999. The limit of detection and limit of quantification were 1.08 and 3.24 μg/ml, respectively. No interference of excipients in determining tablet formulation; identical results were obtained like that of the standard sample.  The proposed RP-HPLC method is simple, accurate, precise, rapid and economical to be employed for routine analysis of carvedilol in pharmaceutical dosage forms. 

An isocratic reverse phase liquid chromatography (RP-LC) method has been developed and subsequently validated for the determination of Lacosamide in Bulk and its pharmaceutical formulation. Separation was achieved with a Xterra RP-8 ((Make: Waters Corporation; 150 mmx4.6 mm I.D; particle size 5 µm)) Column and Sodium di-hydrogen phosphate monohydrate buffer (pH adjusted to 3.0 with diluted orthophosphoric acid): Acetonitrile (800:200) v/v as eluent at a flow rate of 1.0 ml/min. UV detection was performed at 230nm. The method is simple, rapid, and selective. The described method of Lacosamide is linear over a range of 12.0µg/ml to 37.85 µg/ml. The method precision for the determination of assay was below 1.0%RSD. The percentage recoveries of active pharmaceutical ingredient (API) from dosage forms ranged from 99.3 to 100.9%. The results showed that the proposed method is suitable for the precise, accurate and rapid determination of Lacosamide in bulk, its dosage forms. Key Words: Lacosamide, RP-LC, Validation, Dosage form.

  A rapid and sensitive UV-Visible spectroscopic method was developed for the estimation of Ranalozine in pure and its Pharmaceutical formulations. The method was validated as per International Conference on Harmonization [ICH] guidelines. The Ranalozine was monitored at 230nm with UV detection and there is interference of diluent at 230nm for Ranalozine. The method was linear (r2 =0.999) at concentration ranging from 12 to 40μg/ml, precise (intra-day relative standard deviation [RSD] and inter-day RSD values < 1.0%), accurate (mean recovery = 100.2%), specific and robust. The results showed that the proposed method is suitable for the precise, accurate and rapid determination of Ranalozine in bulk, its capsule dosage forms. Key Words: Ranalozine, UV-Visible spectroscopy, Validation, Dosage form.

  This research paper describes simple analytical method for determination of Camylofin dihydrochloride and Paracetamol in syrup formulation by Gas chromatography method. Benzoic acid was used as internal standard. Validation was carried out in compliance with the International Conference on Harmonization guidelines. The method utilized GC (Agilent Technologies 6890N Network GC system with FID detector), and RTX-5 capillary column (Crossbond 50% diphenyl-95% dimethyl polysiloxane), 30m x 0.53mm, 1.5µm as stationary phase. Helium was used as the carrier gas.  The proposed method was validated for linearity, LOD, LOQ, accuracy, precision, ruggedness and solution stability. It can be conveniently adopted for routine quality control analysis. Key Words:Validation, Gas chromatography, Pharmaceutical preparations, Camylofin dihydrochloride, Paracetamol.

  A novel, simple, selective and rugged quantitative method for the determination of Fenofibric acid the active metabolite of fenofibrate  in human plasma (Na2EDTA) using liquid chromatography-tandem mass spectrometric (LC-MS/MS) method has been developed and validated with 200µL human plasma. Fenofibric acid-d6 was used as an internal standard. Analyte and the internal standards were extracted from human plasma by liquid-liquid extraction using Methyl tertiary butyl ether as extraction solvent and ammonium acetate (5mM, pH 2.5) as extraction buffer. The reconstituted samples were chromatographed on a C18 column by using isocratic mobile phase. The method was validated over the concentration range of 79.89–20021.87 ng/mL. The Quattro Premier XE mass spectrometer was operated under the multiple reaction-monitoring mode (MRM) using the electro spray ionization technique for quantification of ion transitions at m/z 317.06/231.00 and 323.24/231.04 for the drug and the internal standard respectively. The method was validated for precision and accuracy, stability, matrix effect, dilution integrity, ruggedness, selectivity and extraction efficiency, and method has been proved to be simple, sensitive, selective, rugged and reproducible. A run time of 2.00 min for each sample made it possible to analyze more than 400 plasma samples per day. The proposed method can be applied for the estimation of the Fenofibric acid in real time plasma samples for pharmacokinetic, drug-drug interaction and toxicological studies.   Key words: Fenofibric acid, Validation, Human Plasma, LC-MS/MS, Electrospray ionization.

  Two simple and sensitive visible spectrophotometric methods (A and B) have been developed for the quantitative estimation of Pioglitazone, in bulk drug and pharmaceutical dosage forms. Methods were based on the formation of Pale yellow coloured and green coloured chromo gens, which were measured 267 nm and 297 nm, respectively. The results obtained with the proposed methods are in good agreement with the labeled amounts when tablet dosage forms were analyzed.  For the first method, UV-spectrophotometry, standard solutions were measured at 267 nm. The first method was linear from 2.5-20 mg/mL. The second method was based on the formation of an ion association complex with Methyl orange (MO) and Bromocresol Green (BCG).The assay was found to be linear over the concentration range of 2.5-20 µg/mL. The wavelengths were selected dependent on the maximum obtained values. The formation of ion- pairs are formed between secondary amino group of PIO and MO, BCG reagents via the protonated nitrogen atom. The proposed methods were validated according to the ICH guidelines (1996) with respect to specificity, linearity, accuracy, precise and robustness. The results demonstrated that the procedure is accurate, precise, specific and reproducible (percent relative standard deviation