Development and Validation of Analytical Spectrophotometric and Rp-HPLC Methods for the Simultaneous Estimation of Hydroquinone, Hydrocortisone and Tretinoin Ternary Mixture in Topical Formulation

Abstract

Objective: Development and validation of spectrophotometric and RP-HPLC methods for the simultaneous determination of Hydroquinone (HQ), Hydrocortisone (HC) and Tretinoin (TRT) ternary combination in pharmaceutical preparation.
 Methods: The proposed spectrophotometric method was able to determine TRT directly from its absorption spectrum at 362 nm, however, HQ and HC from their first derivative spectra at 284 nm and 252 nm, respectively, without any separation step. The RP-HPLC method was developed using a C18 Sunfire© waters column with a mobile phase composed of acetonitrile: phosphate buffer (adjusted to pH 6.1 using ortho-phosphoric acid) in the ratio of 30:70 %, v/v, respectively at a flow rate of 0.8 ml/min. Quantification was based on measuring peak areas at 260 nm.
 Results: The spectrophotometric method was able to selectively quantify each of HQ, HC and TRT in the ranges of 10-50 µg/ml, 2-10 µg/ml and 0.5-5 µg/ml, respectively. The RP-HPLC method was able to produce well-resolved peaks after 3.0, 8.2 and 20.2 min, in the ranges of 2-10 µg/ml, 0.1-1 µg/ml and 0.05-2 µg/ml, for HQ, HC and TRT, respectively. The obtained A, D1 or peak areas values plotted against the concentration of each of the three components showed linear response in the stated ranges. Both methods were validated in terms of linearity, LOD, LOQ, precision, accuracy and selectivity.
 Conclusion: Both developed proposed methods were applied for the determination of the active ingredients in the pharmaceutical formulation and the common excipients did not interfere in the analysis. The RP-HPLC method proved to be more sensitive when compared to the applied spectrophotometric method. However, the applied spectrophotometric methods, considered as green analytical chemistry, is a simple, time-saving method that requires minimal use of a hazardous solvent.

Selective Trace Level Determination of 1,4-Dioxane Content in Sodium Laureth Sulfate Raw Material by Gas Chromatographic-Flame Ionization Detection Method

“…The proposed method validated as per the ICH guidelines [17] and USP Compendia [18] along with the references literature on method validation [19][20][21][22][23][24][25][26][27][28] studies in current industry practices.…”

Trace Level Determination of Sodium Chloride and Sodium Sulfate Content in Sodium Laureth Sulfate Raw Material Using Counter Cation-Exchange HPLC With Indirect Ultraviolet Detection

“…The proposed method validated as per ICH guidelines [20] and USP Compendia [21], along with references literature on method validation [12][13][14][15][16][17][18][19] studies in the current industry practices.…”

“…There is one harmonized method [11] of analysis that is published using high-performance liquid chromatography (HPLC) with charged aerosol detection technique. There was no other selective chromatographic method of analysis for simultaneous [12][13][14][15][16][17][18][19] determination of components of interest that was published in a unique HPLC method with ultraviolet detection. Hence, a method was developed and validated a high-performance liquid chromatographic technique with indirect ultraviolet detection for accurate and precise trace level determination of sodium chloride and sodium sulfate in SLS raw material using counter cation-exchange column.…”

Assay of Tretinoin using Safranine-O as a Chromogenic Reagent in Bulk and Dosage Forms

“…Acne vulgaris is considered as common skin related problem that was treated with combination therapy using topical drug clindamycin and tretinoin (TTN) (1). Literature survey revealed that there is an evidence about the determination TTN by HPLC (2)(3)(4)(5)(6), RP-HPLC (7)(8)(9)(10)(11)(12), UPLC (13), LC (14), and GC (15). Studies on phototoxicity of tretinoin (16), UVspectrophotometry (17)(18)(19)(20)(21)(22), and UV-derivative spectroscopy (23)(24)(25) were also carried out by the previous authors.…”