Development and validation of HPLC method for simultaneous determination of Ketotifen Fumarate and Salbutamol Sulfate in bulk and tablets dosage forms

The aim of the study was to develop and validate a new, rapid, sensitive, simple, accurate and reproducible HPLC method for simultaneous determination of ketotifen fumarate and salbutamol sulfate. Simultaneous HPLC method was developed using RP-C18 stainless steel analytical column 4.6×150 mm C18.Acetonitrile and phosphate buffer pH 4 (30 : 70) were used as mobile phase and wavelength was adjusted to 276 nm for detection of drugs. Developed method was validated for its specificity, accuracy, precision, linearity and robustness. Method was also applied to quantify drugs in commercial tablets. Chromatogram obtained by newly developed method for simultaneous determination of two anti-asthmatic drugs, having well distinguished peaks for both drugs. Retention time of ketotifen fumarate and salbutamol sulfate were 2.69 minutes and 9.47 minutes respectively. Total run time for both anti-asthmatic drugs was 12 minutes. Limit of quantification for ketotifen fumarate and salbutamol sulfate was 1 ng/ml and 1.50 ng/ml respectively. Limit of detection of ketotifen fumarate and salbutamol sulfate was 3.03 and 4.54 respectively. A simple, easy, precise and new method was developed for simultaneous quantification of frequently used anti-asthmatic drugs. Developed method may prove effective and beneficial in determination of ketotifen fumarate and salbutamol sulfate in bulk and other pharmaceutical dosage forms.

Optimization and Evaluation of Orodispersible Solid Dispersion Tablet of Ketotifen Fumarate

The present study was aimed to integrate the developed and optimized ketotifen fumarate dispersion into Orodispersible tablets formulations, to enhance the dissolution rate and bioavailability aspects of the drug. Ketotifen fumarate solid dispersion was prepared using different concentrations of poloxamer 407via solvent evaporation and fusion method. Solubility study, x-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and other investigations were done. Ten formulations of the optimum dispersed ketotifen fumarate Orodispersible tablets were prepared with various superdisintegrants, the results of in vitro - in vivo tests revealed that, the dispersion of the drug in the polymer considerably enhanced the solubility, the batch (Fsd 3) prepared by fusion method showed increased the solubility as ~2-fold compared with a pure drug. FTIR spectra, SEM and XRD data, showed amorphrization of ketotifen fumarate, which explains the better dissolution rate of the drug from its solid dispersions. Formulation F1 containing 15%w/w of crospovidone was showed in vitro- in vivo disintegration time (17 sec., 15 sec. respectively) and percent of drug dissolved in 2 min. was 90.04%, proved to be the optimum formulation, which is required for obtaining rapidly disintegrating tablets. The solubility of the drug had increased, and the resultant orodispersible tablets can be considered as a promising dosage form to achieve better patient compliance.

Mechanistic Studies of the Antiallergic Activity of Phyllanthus amarus Schum. & Thonn. and Its Compounds

Phyllanthus amarus Schum. & Thonn. (Phyllanthaceae) is a medicinal plant that is commonly used to treat diseases such as asthma, diabetes, and anemia. This study aimed to examine the antiallergic activity of P. amarus extract and its compounds. The antiallergic activity was determined by measuring the concentration of allergy markers release from rat basophilic leukemia (RBL-2H3) cells with ketotifen fumarate as the positive control. As a result, P. amarus did not stabilize mast cell degranulation but exhibited antihistamine activity. The antihistamine activity was evaluated by conducting a competition radioligand binding assay on the histamine 1 receptor (H1R). Four compounds were identified from the high performance liquid chromatography (HPLC) analysis which were phyllanthin (1), hypophyllanthin (2), niranthin (3), and corilagin (4). To gain insights into the binding interactions of the most active compound hypophyllanthin (2), molecular docking was conducted and found that hypophyllanthin (2) exhibited favorable binding in the H1R binding site. In conclusion, P. amarus and hypophyllanthin (2) could potentially exhibit antiallergic activity by preventing the activation of the H1 receptor.

Elevated levels of prostaglandin E2 in the tears of patients with severe allergic conjunctivitis and primary cultured conjunctival cells are suppressed by ketotifen and dexamethasone

ObjectiveWe examined the production of prostaglandin E2 (PGE2), which is the key prostaglandin involved in inflammatory disorders of the ocular surface. Tears and conjunctival fibroblasts were evaluated in order to assess allergic inflammation and the effect of specific drugs.Methods and analysisPGE2 was measured in tears from both patients and normal volunteers. Primary cultures of human conjunctival fibroblasts were incubated with interleukin (IL)-4 and tumour necrosis factor (TNF)-α with or without ketotifen fumarate or dexamethasone. The culture supernatants were removed 24 hours after exposure and the concentrations of PGE2 were quantified by ELISA.ResultsSignificantly higher levels of PGE2 were observed in the tears of patients with severe allergic conjunctivitis than in those with post-surgical inflammation (p=0.02), and this production was reduced by eye drops. Stimulation with IL-4 and TNF-α induced the generation of PGE2 in supernatants of conjunctival fibroblasts, and this production was significantly downregulated by ketotifen fumarate or steroids.ConclusionPGE2 may participate in the pathogenesis of severe ocular allergic disease, and both ketotifen fumarate and steroid reduce the production of PGE2.

Determination of Ketotifen Fumarate in Syrup Dosage Form by High Performance Liquid Chromatography

The goal of the current study was to establish and authenticate an isocratic reverse-stage High-Performance Liquid Chromatography (HPLC) method for quantifying ketotifen fumarate (KF) in pharmaceutical liquid dosage compositions. Easy, quick, accurate, exact, and accurate reverse-stage high-performance liquid chromatography was advanced for the simultaneous assessment of ketotifen fumarate in the liquid syrup dosage type. The HPLC system using isocratic elution method with reverse-phase Inertsil ODS-(250 mm × 4.6 mm, 3 μm) column was detected by ultraviolet absorbance at 297 nm with no interference from widely using excipients, the mobile phase (A) is a mixture of triethylamine and water (175 μl in 500 ml of water), and the mobile phase (B) is a mixture of triethylamine and methanol (175 μl in 500 ml of methanol) at a flow rate of 1.5 mL/min (mobile phase A 40 %:mobile phase B 60%) at column temperature using 40 ° C, the retention time for ketotifen fumarate was 6.4±0.5 min. The concentration curves were linear in the range of 10.0 to 35.0 μg / ml (R2 = 0.9999). The developed method was tested for the specificity, precision, linearity, precision, reliability, robustness, and consistency of the solution. The regeneration of ketotifen fumarate in formulations was found to be 99.75 %, 99.91 %, and 100.05 % respectively. The percent RSD for percent recovery was found to be 0.21 and 0.17 and 0.10 for ketotifen fumarate. In the conclusion, the suggested technique was successfully used for the quantitative determination of ketotifen fumarate in formulations.