SIMULTANEOUS ESTIMATION OF AZILSARTAN AND CILNIDIPINE IN BULK BY RP-HPLC AND ASSESSMENT OF ITS APPLICABILITY IN MARKETED TABLET DOSAGE FORM

Objective: This study aims to build up the RP-HPLC process for Azilsartan and Cilnidipine and authenticate the RP-HPLC process according to ICH validation code Q2R1. Methods: System suitability testing was performed to discover the qualifying criterion of the method by injecting the identical standard solution of Azilsartan 40μg/ml and Cilnidipine 10μg/ml in mixture/combination in subsequent optimized chromatographic conditions and the chromatogram was recorded. Moreover, the planned method was validated as per ICH guideline Q2R1 for the following parameters: linearity and range, precision, accuracy, robustness, and determined % recovery. Results: The outcomes of %RSD for retention time and peak area were found to be 0.65 and 1.32 for Azilsartan and 0.85 and 1.90 for Cilnidipine. The correlation coefficient, y-intercept, slope of the regression line were 0.9996,-1127.1, 3313.9, and 0.9993, 1460.2, 2876.4 for Azilsartan and Cilnidipine, respectively. Moreover, the range of this method was observed to be 40-240μg/ml and 10-60 μg/ml for Azilsartan and Cilnidipine, standard concentrations respectively. The % RSD achieved for precision (repeatability) was observed in the range of 1.57 to 2.43 for Azilsartan and 0.70 to 1.88 for Cilnidipine. The % accuracy was found in the range of 96.96 to 101.92% w/w for Azilsartan and 99.19 to101.96%w/w for Cilnidipine. The percent recovery values achieved for Azilsartan were in the range of 99.87 to 106.39% w/w and for Cilnidipine in the range of 94.51 to 105.96% w/w. Conclusion: The author concludes that the simultaneous estimation of Azilsartan and Cilnidipine with predefined objectives was successfully achieved. Moreover, the method was found to be steadfast for the quantification of Azilsartan and Cilnidipine in marketed tablet dosage forms.

VALIDATED NOVEL SPECTROSCOPIC METHOD FOR ESTIMATION OF TRIAMCINOLONE ACETONIDE IN NASAL SPRAY

A simple, sensitive, accurate, precise and cost-effective pH independent spectrophotometric method has been developed for the estimation of triamcinolone acetonide in nasal sprays. Estimation was carried out at the isosbestic point of drug solutions, Isosbestic point was measured by scanning equimolar solution of triamcinolone acetonide separately in phosphate buffer pH 3.0, phosphate buffer pH 7.0 and phosphate buffer pH 10.0 at a concentration of 10 µg mL-1. They were scanned in the wavelength range of 200-400 nm. The isosbestic point was found to be 230.0 nm in acidic, basic and neutral condition; hence pH independent wavelength 230 nm was used for estimation of triamcinolone acetonide. The method observed linear response in the concentration range of 3-18 µg mL-1 at 230 nm. Developed method has been validated in accordance with ICH guideline Q2 R1 for several parameters like linearity, precision, accuracy, limit of detection and limit of quantification. The developed and validated analytical method was successfully applied to the estimation of triamcinolone acetonide in marketed nasal spray formulations.

Lawsone Co-Crystals Loaded Antifungal Gel for Cutaneous Candidiasis

Aim: An attempt was made to improve the solubility and to achieve better penetration of antifungal agent lawsone a poorly water-soluble naphthoquinone, derivative via co-crystallization to treat cutaneous candidiasis. Methodology: The co-crystals of lawsone were prepared using benzoic acid as co former by solvent crystallization method. The formulated co-crystals were assessed for different parameters namely, saturation solubility, thermal properties, crystalline nature, particle size and its antifungal activity against Candida albicans. Results: Lawsone co-crystals exhibited enhanced solubility of lawsone (9.57±2.5 mg/ml and 0.523±0.23 mg/ml respectively) and the particle size of the co-crystals were reduced to 560±2.2 nm as compared to pure lawsone (2478±1.5 nm) which further resulted in enhancement of antifungal activity. Lawsone co-crystals were loaded in the xanthan-gel base for easy applicability to the skin and to achieve patient compliance. The prepared gel was evaluated in terms of spreadability, adhesiveness, in vitro diffusion, viscosity, and antifungal activity. Conclusion: Lawsone co-crystals loaded gel showed enhanced retension of drug in the skin as compared to plain lawsone gel. The antifungal potential of lawsone co-crystals loaded gel was at par with marketed Clotrimazole gel formulation. The short-term stability study carried out as per ICH guideline indicated that the formulation was stable. Lawsone co-crystals loaded gel.

ANALYTICAL QUALITY-BY-DESIGN (AQBD) APPROACH FOR THE DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE ESTIMATION OF LAMOTRIGINE IN BULK AND TABLET FORMULATION

The current analytical exploration illustrated developing a reversed-phase high-performance liquid chromatography (RP-HPLC) technique and consequent substantiation for analyzing lamotrigine (LAM) active pharmaceutical ingredient (API) using a Quality-by-design (QbD) approach (Central Composite Design), in bulk product as well as in the tablet formulations. In this experiment, based on systematic scouting, four key components (viz., mobile phase, column, flow-rate, and wavelength) were studied by the RP-HPLC method. 13 experimental runs were done with acetonitrile (ACN) (40-60% v/v) having flow-rate in the range 0.8 mL/min to 1.2 mL/min. The proposed analytical method was thoroughly corroborated in terms of ruggedness linearity, robustness, accuracy, and precision in accordance with ICH guideline Q2A and ICH guideline Q2B. Under the optimum chromatographic environment; Intersil C8 column of 250 mm length, 4.6 mm (i.d.); 20 μL injection volume; and mobile phase ACN: Methanol (60:40 v/v), a retention time of 2.542 min was noticed at 220 nm detection wavelength. The method was found to be extremely reproducible, accurate, linear, precise, robust, and economically adequate to execute the estimation. The intended analytical technique was thoroughly assessed through statistical tools and could be an imperative concern for the habitual scrutiny of LAM in bulk products and its formulation.

Bioanalytical Method Development and Validation of Dapoxetine Hydrochloride in Human Plasma by RP-HPLC

A simple, accurate and rapid Bioanalytical reverse phase high performance liquid chromatography (RPHPLC) method for determination of Dapoxetin hydrochloride in human plasma was validated as per ICH guideline. Dapoxetin hydrochloride is significantly superior in premature ejaculation and more active against serotonin transport inhibitor than any other drug in class. The total analysis was carried out on using stationary phase symmetry C1 (4.6mm X 250mm, 5µm) with Mobile Phase Acetonitrile: Buffer (60:40) pH adjusted to3.5 flow rate was 1.0 ml/min, injection volume of 10 ppm and detection wavelength was 293nm at ambient temperature with total run time of 10 minutes. Retention time of spiked plasma and dapoxetine hydrochloride were found to be 2.153 min and 4.442 min, r2 value were 0.995 and 0.999 and linearity range was 5ppm to 25ppm for both. The method was developed for accuracy, linearity, precision, recovery and stability in complies and stability in complies with CDER and ICH guideline.

Development and Stability of Forced Degradation Studies Indicating Different Brands of Metformin Drugs

In present study, Accouring to specification of Indian pharmacopeia the content official limit of not less than (98.5%) and not more than (101.0%) of the lable amount our hypothesis was that when all different brands of metformin were expose to the different degradation parameters. The Forced degradation studies show the chemical behavior of the molecule which in turn helps in the development of formulation and package. A forced degradation study is an essential step in the design of a regulatory compliant stability program for both drug substances and products, and formalized as a regulatory requirement in ICH Guideline Q1A in 1993. Forced degradation is a degradation of new drug substance and drug product at conditions more severe than accelerated conditions.

Characterization and In-silico toxicity prediction of degradation products of felbamate

Background The objective of the work carried out was to assess the toxicity of the degradation products (DPs) for the drug felbamate. Stress studies were performed in the condition specified in the international council of harmonization (ICH) guideline Q1A (R2). Results The drug degraded under the alkaline stress conditions to generate two degradation products (DPs). They were separated on a Phenomenex C8 column (250 mm × 4.6 mm, 5 µm); mobile phase composition was 10 mM ammonium formate (pH adjusted to 3.7 with formic acid) and acetonitrile (80:20, v/v); flow rate and wavelength for recording absorbance were 1.0 ml/min and 206 nm, respectively. The structures of the degradation products were characterized by LC–MS/MS analysis. Conclusion The drug was prone to hydrolysis in the presence of alkali. It was found to be stable under other stress conditions, viz., acidic, neutral, thermal, photolytic and oxidative. The structures of the impurities were characterized by LC–MS/MS. The drug and the DPs were screened through ADME and toxicity prediction software’s like pkCSM, Toxtree and OSIRIS property explorer. Felbamate was flagged for possible hepatotoxicity.

Development and Validation of TLC of Flavonoid from the Ethanolic Extract of Plant Enhydra fluctuans

Thin layer chromatography is a technique or an analytical tool to separate the bioactive compound from the mixture of components. In current research work special attention was given to develop specific solvent system and to validate the principle of separation of Flavonoid. The ethanolic extract of plant namely Helencha (in Bengali) was selected for such purpose. After several trials, the presence of Flavonoid which was confirmed by qualitative evaluation and was sepahjvvrated successfully under this study and the process was validated under the circumstances of ICH Guideline. The plant not only contained Flavonoid but there were the presence of little quantity of Alkaloid, Saponin and Tannins also. Due to presence of Flavonoid the ethanolic fraction of the plant may be evaluated for Anti-inflammatory and Anthelmintic activity for further research. The plant Helencha is known as Enhydra fluctuans belongs to the family Astereceae. According to folklore claim the plant is useful for nutrition purpose. Not only that the plant is also useful in Dropsy, anasarca and snake bite. As per the literature survey, the plant has Antioxidant and Analgesic activity. Here the total attention was given to separate Flavonoid from the mixture of Components present in the ethanolic fraction of the leaves of the plant. Keywords: Flavonoid, Alkaloid, ICH Guideline, Enhydra fluctuans, Dropsy, Anaasrca.

Development, Validation and Application of an ICP-SFMS Method for the Determination of Metals in Protein Powder Samples, Sourced in Ireland, with Risk Assessment for Irish Consumers

A method has been developed, optimised and validated to analyse protein powder supplements on an inductively coupled plasma-sector field mass spectrometer (ICP-SFMS), with reference to ICH Guideline Q2 Validation of Analytical Procedures: Text and Methodology. This method was used in the assessment of twenty-one (n = 21) elements (Al, Au, Ba, Be, Bi, Cd, Co, Cr, Cu, Fe, Hg, Li, Mg, Mn, Mo, Pb, Pt, Sn, Ti, Tl, V) to evaluate the safety of thirty-six (n = 36) protein powder samples that were commercially available in the Irish marketplace in 2016/2017. Using the determined concentrations of elements in samples (µg·kg−1), a human health risk assessment was carried out to evaluate the potential carcinogenic and other risks to consumers of these products. While the concentrations of potentially toxic elements were found to be at acceptable levels, the results suggest that excessive and prolonged use of some of these products may place consumers at a slightly elevated risk for developing cancer or other negative health impacts throughout their lifetimes. Thus, the excessive use of these products is to be cautioned, and consumers are encouraged to follow manufacturer serving recommendations.

DEVELOPMENT OF LIQUID CHROMATOGRAPHIC METHOD FOR ESTIMATION OF ACAMPROSATE CALCIUM AND BACLOFEN COMBINATION USED IN TREATMENT OF NEUROLOGICAL DISORDERS

A specific, accurate, precise, and reproducible liquid chromatographic method has been developed and validated for the estimation of acamprosate calcium and baclofen in combination. The separation was achieved using stationary phase Phenomenex C18 column (150 mm× 4.6 mm.) in isocratic mode, with mobile phase containing 0.05 M potassium dihydrogen orthophosphate buffer (pH 7) : acetonitrile (10:90 V/V), at a flow rate of 1.0 mL/min and effluents were monitored at 210 nm. The retention time of acamprosate calcium and baclofen were found to be 1.9 min and 5.3 min, respectively. The linearity for acamprosate calcium and baclofen were in the range of 2-64 µg/mL and 1.2- 38.4 µg/mL, respectively. The method was validated as per ICH guideline. The recoveries of Acamprosate calcium and baclofen were found in the range of 98.90 - 100.13 % and 98.60 -100.02 %, respectively. The method was successfully applied for the determination of both the drugs in combination.