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.


[1]       A. Jakimska, B. Huerta, Z. Barganska, A. Kot-Wasik, S. Rodriguez-Mozaz, D. Barcel, J. Chromatogr. A 1306 (2013) 44.
[2]       C. Hu, M. He, B. Chen, Ch. Zhong, B. Hu, J. Chromatogr. A 1310 (2013) 21.
[3]       L. Barreiros, J.F. Queiroz, L.M. Magalhaes, A.M.T. Silva, M.A. Segundo, Microchem. J. 126 (2016) 243.
[4]       J. Liu, Q, Luo, Q. Huang, J. Clean. Prod. 139 (2016) 1400.
[5]       A. Hom-Diaz, M. Llorca, S. Rodriguez-Mozaz, T. Vicent, D. Barcel, P.B. anquez, J. Environ. Manage. 155 (2015) 106.
[6]       A. Gonzalez; J. Avivar, V. Cerda, J. Chromatogr. A 31 (2015) 1.
[7]       L. Zelena, J. Reznicek, M. Ceckova, H. Sklenarova, J. Pharm. Biomed. Anal. 137 (2017) 70.
[8]       R.C. Schofield, D.R. Mendu, L.V. Ramanathan, M.S. Pessin, D.C. Carlow, J. Chromatogr. B 1048 (2017) 70.
[9]       M. Chen, Y. Liu, Ch. Lin, V.K. Ponnusamy, J. Jen, Anal. Chim. Acta 767 (2013) 81.
[10]    X. Li, Sh. Li, G. Kellermann, Talanta (2018) 1.
[11]    R. Zhang, N. Li, Ch. Wang, Y. Bai, R. Ren, Sh Gao, W. Yu, T Zhao, H. Zhang, Anal. Chim. Acta 704 (2011) 98.
[12]    P.C.F.L. Gomes, B.B. Barnes, A.J. Santos-Neto, F.M. Lancas, N.H. Snow, J. Chromatogr. A 1299 (2013) 126.
[13]    Sh.X.L. Goh, H.K. Lee, J. Chromatogr. A (2017) 1.
[14]    C. Bicchi, T. Schiliro, C. Pignata, E. Fea, C. Cordero, F. Canale, G. Gilli, Sci. Total Environ. 407 (2009) 1842.
[15]    K.  Yuan,  H.  Kang,  Zh.  Yue,  L.  Yang,  L.  Lin,  X.
Wang, T. Luan, Anal. Chim. Acta (2015) 1.
[16]    M. Torbati, M.A. Farajzadeh, M. Torbati, A.A. Alizadeh Nabil, A. Mohebbi, M.R. Afshar Mogaddam, Talanta (2017) 1.
[17]    T.L. Chen, Sh. H. Tzing, W.H. Ding, J. Chromatogr. A (2015) 1.
[18]    S. Song, E.N. Ediage, A. Wu, S.D. Saeger, J. Chromatogr. A 1292 (2013) 111.
[19]    K. Cherkashina, Ch. Vakh, S. Lebedinets, A. Pochivalov, L. Moskvin, A. Lezov, A. Bulatov, Talanta (2018) 1.
[20]    A. Pourhossein, K. Alizadeh, J. Reports Pharm. Sci. 7 (2018) 79.
[21]    M. Bahmaei, H.A. Mashayekhi, F. Khalilian, J. Braz. Chem. Soc. 26 (2015) 1475.
[22]    Ch. Ch. Chang, Sh. D. Huang, Anal. Chim. Acta 662 (2010) 39.
[23]    G.D. Orazio, J.H. Borges, A.V.H. Herrera, S. Fanali, M.A.R. Delgado, Anal. Bioanal. Chem. 408 (2016) 7447.
[24]    H. Feng, L. Ning, L. Xiao-Li, J. Northeast Agric. Univ. 23 (2016) 44.
[25]    X. Wang, P. Huang, X. Ma, X. Du, X. Lu, Talanta 194 (2019) 7.
[26]    R.C. Schofield, D.R. Mendu, L.V. Ramanathan, M.S. Pessin, D.C. Carlow, J. Chromatogr. B 1048 (2017) 70.
[27]    A.A. Lahcen, A.A. Baleg, P. Baker, E. Iwuoha, A. Amine, Sens. Actuators B 241 (2017) 698.
[28]    C. Hu, M. He, B. Chen, Ch. Zhong, B. Hu, J. Chromatogr. A 1310 (2013) 21.
[29]    L. Patrolecco, N. Ademollo, P. Grenni, A. Tolomei, A.B. Caracciolo, S. Capri, Microchem. J. 107 (2013) 165.