Development and Validation of HPTLC Assay Method for Simultaneous Quantification of Hydrocortisone and Clotrimazole in Cream and Applying for Stability Indicating Test

Abstract

A simple, specific, precise and accurate simultaneous high-performance thin-layer chromatographic method of analysis for hydrocortisone and clotrimazole was developed and validated. The method employed HPTLC glass plates (20x10) mm precoated with silica gel 60 F 254 as the stationary phase. The solvent system consisted of Toluene: Propanol: Ammonia (13:3:0.1v/v) that gave compact spots with R f value of 0.27 ± 0.01 and 0.58 ± 0.02 for hydrocortisone and clotrimazole, respectively. Densitometric analysis of hydrocortisone and clotrimazole was carried out in the absorbance/reflectance mode at 226nm. The calibration curves showed good linear relationship with R 2 = 0.996 + 0.003 and 0.996 + 0.002 in the concentration range of 200-1200 and 200-1000 ng/µl for hydrocortisone and clotrimazole, respectively. The method was validated for precision, recovery and robustness. The LOD & LOQ were found to be 35.31, 107.01 and 34.93, 105.87 ng/µl for hydrocortisone and clotrimazole respectively. Statistical analysis proved that the method is repeatable, specific and accurate for the estimation of the studied drugs. Resolution of hydrocortisone, clotrimazole and their degradation products formed under different stress conditions (acid-base hydrolysis, oxidation and thermal degradation) was successfully achieved with the developed method indicating that it can be employed as a stability indicating method. Hydrocortisone (cortisol);11,17,20-dione, the main physiologic glucocorticoid in humans has an anti-inflammatory and an immunosuppressive effect besides its role on carbohydrate and protein metabolism 1-3 . The anti-inflammatory effect of hydrocortisone is due to reduction of histamine release, reduction of IgG production and decreasing of the activity of neutrophils and macrophages [4][5][6]imidazole is an imidazole derivative with antimycotic activity that is known to inhibit cytochrome P-450, ergosterol biosynthesis, and proliferation of cells and to interfere with cellular Ca 2+ homeostasis 7-9 . BACKGROUND

Identification and Qualitative Analysis of β-Sitosterol and Some Phytoestrogens in In Vivo and In-Vitro Samples of Lepidium sativum: A Semi-Arid Bone Healing Plant

“…In the present investigation Chloroform: methanol (8:2) combination showed best results and exhibited coherence with the results of [13,14]. HPTLC technique with other biochemical assays supplemented evidence to confirm the presence of some bioactive compounds, which have provided the plants an important position in folkloric usage [15] In the present study, quantitative and qualitative biochemical methods can be employed as a stability indicator [16] that can be used for the routine quality control analysis and quantitative determination of β-sitosterol from Lepidium sativum. Results obtained by HPTLC for diazdzein and formononetin were coincided with the results of [17][18][19].…”

Analytical method development and validation of hydrocortisone and clotrimazole in topical dosage form using RP-HPLC

“…Literature survey disclose that different methods are available for detection of HYD and CL separately; for HYD, these are polarographic [12,13], HPLC [14,15], spectrophotometric [16], TLC [17], and LC-MS/MS [18,19] methods; and for CL, these are spectrophotometric [20], HPLC methods [21,22], and titrimetric [23] TLC [24] methods. Only one method has been reported for the simultaneous determination of HYD and CL that is HPTLC [25]. The aim of this work was to develop and validate a new, novel, and different method for the simultaneous determination of HYD and CL in a topical cream and in a raw form.…”

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

“…Few reverse-phase highperformance liquid chromatographic (RP-HPLC) methods were reported for the estimation of hydrocortisone alone and with other combinations [4][5][6][7]. A high-performance liquid chromatographic tandem mass spectrometric method for the estimation of hydrocortisone in human urine has been reported [8] and a highperformance thin-layer chromatographic method has been reported for quantification of hydrocortisone and clotrimazole [9]. Along with these RP-HPLC methods have reported for the estimation of tetracycline [10,11].…”