| Summary: | Abstract Accurate quantification of steroid hormones based on LC–MS/MS is of great significance for the diagnosis of endocrine diseases and the study of metabolomics. However, the current liquid–liquid extraction (LLE) and solid-phase extraction (SPE) are difficult to be further accepted in the pretreatment of clinical samples. Therefore, we carefully developed and optimized a novel method for the treatment of human serum samples by magnetic solid-phase extraction (MSPE). Several necessary tests for 12 steroids, such as LLOQ, matrix effect, recovery and precision, have verified the repeatability and practicability of the method. The comparison with LLE proves that the environment-friendly MSPE has the significance of further popularization in the treatment of clinical samples. Focusing on the future, the study opens the door for the automatic preprocessing of human serum samples based on MSPE, including but not limited to the quantitative analysis of steroid hormones.
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