Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Solid-phase extraction combined with high performance liquid chromatography-diode array detector for rapid wetermination of estrogens in milk

Qinghua Yan¹, Li Yang², Sufang Li³

¹Department of Life Science and Technology, Xinxiang Medical University; ²Department of Experimental Center, Henan Institute of Science and Technology.

For correspondence:- Sufang Li Email: yqh3499@163.com Tel:+863733831928

Received: 20 July 2015 Accepted: 12 October 2015 Published: 29 November 2015

Citation: Yan Q, Yang L, Li S. Solid-phase extraction combined with high performance liquid chromatography-diode array detector for rapid wetermination of estrogens in milk. Trop J Pharm Res 2015; 14(11):2077-2082 doi: 10.4314/tjpr.v14i10.18

© 2015 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To develop a simple but effective method for the quantitative analysis of estrogens in milk.

Methods: Solid-phase extraction method was employed for the extraction of the estrogen from milk and high performance liquid chromatography-diode array detector (HPLC-DAD) was used for the determination of estrogen.

Results: Optimal chromatographic conditions were achieved on an Eclipse XDB-C18 column at a flow rate of 0.8 mL/min at room temperature. The products were monitored using a DAD detector set at 280 nm. The retention time of the three main estrogens were 15 min with a gradient program of acetonitrile/water, from 0 to 3 min, holding 35 % acetonitrile; from 4 to 7 min, ramped to 55 % acetonitrile; from 8 to 20 min, maintained at 55 % acetonitrile. The results showed that the calibration curve had good linearity within the concentration range of 0.5 - 8.0 µg/mL with correlation coef@257;cient (R²) in the range of 0.9929-0.9936. The limit of detection was in the range of 0.025 - 0.045 µg/mL while mean recovery of estrogen from milk samples varied from 92.6 to 104.1 %. Satisfactory precision was obtained both for intra-assay (RSD, 1.8 to 4.4 %) and inter-assay (RSD, 2.0 to 4.7 %).

Conclusion: The proposed method is environmentally friendly, inexpensive and convenient, and should be helpful in analyzing estrogens in biological, environmental and food samples.

Keywords: Solid-phase extraction, Milk, Estrogens, High performance liquid chromatography-diode array detector