Abstract
A rapid, sensitive and specific reversed-phase high performance liquid chromatographic method with diode-array detection has been developed and validated for the determination of hydroxybenzene (0.494%, w/w) in a commercially available cream pharmaceutical formulation. Isocratic chromatography was performed on a C18 column with methanol-water 60:40 (v/v) containing 0.1% phosphoric acid (v/v) as mobile phase at a flow rate of 1.0 ml/min. UV detection was at 254 nm. Linearity of the method was excellent (r2 = 0.9999). The relative standard deviation values for intra- and inter-day precision studies were < 1% and the recovery of hydroxybenzene was >99%. The limit of detection and quantitation for hydroxybenzene was found to be 13.5 η g/ml and 2 μg/ml, respectively. The method was also validated for specificity and robustness. The method was found to be robust and can be reliably used to determine the hydroxybenzene content of marketed formulations.
Highlights
Hydroxybenzene is widely used in the preparation of drugs, cosmetics including sunscreens[1], hair dyes, and skin lightening preparations[2]
RP-HPLC is the opposite of normal-phase chromatography, with a nonpolar stationary phase and a polar, largely aqueous mobile phase
The flow rate was constant at 1.0 ml/min and the column temperature was at room temperature (24±1 )
Summary
Hydroxybenzene is widely used in the preparation of drugs, cosmetics including sunscreens[1], hair dyes, and skin lightening preparations[2]. Hydroxybenzene belong to a family of compounds especially harmful for the environment, owing to their toxicity and carcinogenic effect[3]. RP-HPLC is the most popular analytical technique for separating complex mixtures in the chemical, pharmaceutical and biotechnological industry. Silica has a small pH range (3 to 8) where mixtures can be separated without degradation of the column performance. The separation is achieved by analytes having different interactions with the stationary phase. In RP-HPLC, solutes are separated using their hydrophobicity. Water is considered to be the weak component of the mobile phase and does not interact with the hydrophobic stationary phase chains. Specificity, linearity, precision (repeatability and intermediate precision), accuracy, robustness, limit of detection and limit of quantitation were evaluated
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