Novel Novel Stability-Indicating High-Performance Liquid Chromatographic Assay Method for Simultaneous Estimation of Hydrocortisone and Tetracycline in Bulk and Pharmaceutical Dosage Form

Abstract

Objective: A combination of hydrocortisone and tetracycline as topical ophthalmic ointment is used for skin irritations, eye infections, inflammation, skin infections, acne, and rashes. The objective of the current work is to a simple, rapid, accurate, and precise, stability-indicating reverse-phase liquid chromatographic method was developed for the simultaneous estimation of hydrocortisone and tetracycline in bulk and pharmaceutical dosage form.
 Methods: The separation was carried out in Discovery C18 column (250 × 4.6 mm, 5 μm) using mobile phase ratio of water (pH 2.2 adjusted with orthophosphoric acid):acetonitrile (40:60 v/v) in an isocratic elution mode with a flow rate of 1.0 ml/min at detection wavelength of 244 nm. The injection volume was 10 μl and the column temperature was set at 30°C.
 Results: The retention time for hydrocortisone and tetracycline was found to be 2.214 ± 0.001 min and 3.497 ± 0.001 min, respectively. Calibration curves were linear (r2=0.999) at a concentration range of 2.5–15 mg/ml for both hydrocortisone and tetracycline. The percentage recoveries were found to be 99.13–99.67% for hydrocortisone and 99.39–99.61% for tetracycline. Relative standard deviation was found to be 0.3% for both the drugs. Limit of detection and limit of quantification values of hydrocortisone and tetracycline were found to be 0.09 μg/ml and 0.27 μg/ml and 0.17 μg/ml and 0.52 μg/ml, respectively. The drugs were subjected to various stress conditions and found no interference of degraded products peak at the retention times of analyte peaks.
 Conclusion: A rapid and accurate reverse-phase high-performance liquid chromatographic method was developed for simultaneous estimation of hydrocortisone and tetracycline, and the method was validated as per the International Council for Harmonization guidelines. Hence, the developed method can be successfully applied for the simultaneous estimation of hydrocortisone and tetracycline in bulk and ointment formulation.

Stability indicating high performance thin layer chromatography method development and validation for quantitative determination of tetracycline hydrochloride in tetracycline hydrochloride active pharmaceutical ingredient (API) and its dosage forms

“…Peak area and amount of drug per band were correlated in the polynomial regression equation with a determination coefficient of 0.9999 (Table 3 ). The linearity of the current developed method (r 2 = 0.9999) was observed to be better than that of the HPLC analytical method, which had a linearity range of 10–150 μg/mL and r 2 of 0.9986 [ 40 ] and an r 2 value of 0.999 at concentrations between 2.5 and 15 mg/mL [ 25 ] for the determination TC-HCl in an ointment of tetracycline HCl (Additional file 1 : Fig. S7).…”

“…Various analytical methods have been documented to identify and assay tetracycline HCl in different formulations. The methods reported include microbiological assay [ 14 ], thin layer chromatography [ 15 ], UV–visible spectroscopy [ 16 – 19 ], flame atomic absorption spectroscopy, infrared spectroscopy [ 20 ], electroanalytical methods [ 21 – 23 ], and HPLC [ 20 , 24 , 25 ]. Official methods for identification and assay are based on infrared spectroscopy, UV–visible spectroscopy, paper chromatography, thin layer chromatography (TLC), and liquid chromatography (LC) techniques [ 26 ].…”