Simultaneous Determination of Hormones in Botanical Dietary Supplements With Supramolecular Solvent-Based Extraction by HPLC-MS/MS

Abstract

Dietary supplements typically contain either natural products or synthetic chemicals. While botanical dietary supplements are often thought to be more natural and safer, they could still contain potentially harmful ingredients. Despite the detection of sex hormones in plants and soil, it is still worth examining if products made from plants contain them. In this study, simultaneous determination of 14 kinds of sex hormones in botanical dietary supplements was conducted by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). A suitable sample pretreatment method is expected to extract and purify the sample in a single step when detected by HPLC-MS/MS, enabling it to be directly injected into the instrument. Herein, supramolecular solvent (SUPRAS), composed of n-hexanol, tetrahydrofuran (THF), and water, was employed to extract hormones from botanical dietary supplements. The effect of various parameters, including the composition of the supramolecular solvents, their dosage, and vortex time, on extraction efficiency was investigated. The results demonstrated that the 14 compounds exhibited excellent linearity over the concentration range of 0.5–100 µg·L⁻¹, with determination coefficients exceeding 0.99. Recoveries were assessed at three levels using spiked samples, ranging from 75.6% to 115.8%, with relative standard deviations between 0.8% and 9.7%. This method was successfully applied to determine 14 sex hormones in botanical dietary supplements. Progesterone was detected in one sample at a concentration of 64 µg·kg⁻¹. The SUPRAS method coupled with HPLC-MS/MS, was proved to be efficient, easy-to-operate, and sensitive for determining sex hormones in botanical dietary supplements.