Online solid phase extraction with liquid chromatography-tandem mass spectrometry for determination of estrogens and glucocorticoids in water

- An automated online SPE procedure was developed to extract steroid hormones.
- Six estrogens and six glucocorticoids were studied.
- Improved analytical performance was achieved.
- Low sample volume and minimal analysis time were attained.
- Isotopic standards helped to minimise matrix effects.

The present work describes the development of a novel fully automated online solid phase extraction (SPE) coupled with high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) using negative electrospray ionisation (ESI) for the simultaneous determination of six estrogens and six glucocorticoids in water. Filtered water samples (5 mL) were preconcentrated on a HyperSep™ Retain PEP SPE cartridge, eluted in back-flush mode, and separated on an LC column before analysis by tandem mass spectrometry. The total analysis time for each sample was 17 min. Different experimental parameters such as the type of online SPE cartridge, loading flow rate, and composition of methanol in the loading phase were optimised. The intra-day repeatability of method ranged from 1.48 to 9.68% for all analytes, and the inter-day reproducibility ranged from 2.03 to 8.63% for all analytes, except for dexamethasone at 11.95%. These were calculated based on the peak area responses of the targeted analytes spiked at 50 ng/L in ultrapure water. The method also showed good linearity from 1 to 100 ng/L, with the limits of detection (LODs) ranging from 0.16 to 2.14 ng/L. The proposed method was applied to the analysis of municipal wastewater. This fully automated online SPE extraction coupled with LC-MS/MS method is effective and reliable to measure estrogens and glucocorticoids simultaneously due to its high throughput, relatively low solvent consumption, reusability of the online SPE cartridge, and reduction of manual labor.