Simultaneous determination of estrogens and progestogens in honey using high performance liquid chromatography-tandem mass spectrometry

Authors: MA, LI - University Of California; ASHWORTH, DANIEL - University Of California; Yates, Scott

Submitted to: Journal of Pharmaceutical and Biomedical Analysis
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/1/2016
Publication Date: 9/3/2016
Citation: Ma, L., Ashworth, D., Yates, S.R. 2016. Simultaneous determination of estrogens and progestogens in honey using high performance liquid chromatography-tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis. 131:303-308. https://doi.org/10.1016/j.jpba.2016.09.001.
DOI: https://doi.org/10.1016/j.jpba.2016.09.001

Interpretive Summary: Concern has been raised over steroidal hormones in environment and agricultural systems. The adverse effects from exposure to hormones include intersexuality, masculinization, and reproductive and behavioral problems. But, hormones that pass through wastewater treatment systems and those associated with concentrated animal feeding operations can enter surface water, which poses potential risks to wildlife and humans. Furthermore, as hormonal chemicals move through the environment, they may enter the food chain from use of reclaimed or impacted water for irrigation or from the use of hormone-containing fertilizers. Thus, there is an urgent need for highly accurate and reliable methods to measure the concentrations of steroidal hormones in soil, water, plant and food samples. Due to its beneficial nutritional properties, honey is consumed worldwide. But, a number of studies have demonstrated the presence of various endocrine disrupting chemicals, pesticides, and antibiotics in honey, highlighting a potential need to protect consumers. Since there is limited information on the presence of steroidal hormones in honey, a new method to measure hormones was developed and the method was tested by sampling four sources of honey for the presence of 13 hormonal chemicals commonly found in the environment. The results show that only 2 of the 52 samples had measureable concentrations and both were near the method’s detection limit (i.e., trace levels). This new method provides a tool for measuring hormones in the environment and the food supply, and can be used in an effort to develop new strategies mitigate hormone contamination. The new measurement methodology and the test results would be of interest to researchers, regulators, commodity groups, farmers, and the public.

Technical Abstract: This work describes the development and validation of a method for the simultaneous determination of 13 estrogens and progestogens in honey by high performance liquid chromatography-tandem mass spectrometry. The target compounds were preconcentrated by solid phase extraction. Pretreatment variables were optimized for a better compatibility with electrospray ionization interfaced mass spectrometry. The analytes were analyzed in multiple-reaction-monitoring mode with two pairs of precursor product ion transitions. The proposed method was validated with method detection limits of 0.01–0.33 ng/g and good linearities (r2 > 0.99) throughout the studied concentration range. The recoveries of analytes at the spiking levels (5 ng/g and 25 ng/g) ranged from 71.2% to 99.7%, with relative standard deviations below 20%. The method was used to determine the target compounds in honey samples (orange blossom, clover and multiflower) obtained from supermarkets. Two samples of honey were found to contain trace amounts of estrone (< MQL) or progesterone (0.2 ng/g).