Determination of 17-β-Estradiol in Water Samples Using Salting-out Assisted Liquid-liquid Extraction Followed by HPLC and Experimental Design for Optimization

Document Type : Research Paper


1 Department of Chemistry, Lorestan University, Khorramabad 6813717133, Iran

2 Department of Chemistry, Tabriz University of Medical Sciences, Tabriz 1476651664, Iran


The aim of the present study was to extract, preconcentrate and determine 17-β–Estradiol in water samples using a simple and efficient method; to this aim, salting-out-assisted liquid-liquid extraction and high-performance liquid chromatography with ultra violet detection at 210 nm were performed. Water-miscible acetonitrile, as the extractant and acetonitrile phase separation under high-salt conditions were applied to treat the samples. The extraction efficiency and method sensitivity were carefully monitored by controlling the effective factors and the optimum conditions were: Sodium chloride as the salting-out agent at concentration of 1.6 g, 2.40 mL of acetonitrile as extraction solvent, 5 mL of water sample, vortexing for 2 minutes and centrifuging at 4000 rpm for 5 minutes. A central composite design was applied to optimize the hydrolysis parameters. Using optimized experimental conditions, the calibration curve was found to be linear in the range of 1–120 µg L-1 in water sample and the correlation coefficient (R2); the limit of detection; limit of quantification were >0.99, 0.25 µg mL-1 and 0.83 μgL-1, respectively. The enrichment factor and extraction recoveries of the selected analyte ranged from 44.96 to 49.57 and 89.93 to 99.15% respectively. Relative standard deviations are about 5.94%. High extraction efficiency and compatibility with HPLC analysis of 17-β–Estradiol in water samples are the advantages of this method.


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