17β-Estradiol (E2) is an endogenous steroid in the human body. Its measurement is important for health and human biology understanding. However, E2 concentration in human plasma is in the range of pg/mL, which makes it difficult to detect. In this way, LC–MS/MS has been shown the most sensitive tool, although E2 is a weakly ionizable molecule. In this work, we validated a more sensitive and accurate method for E2 quantification in human plasma. Our extraction step ensured a cleaner chromatography, resulting in a precise measurement and highly reproducible method in the range of 2–150 pg/mL. Moreover, we proved a long stability for E2 in several conditions. All results indicate that our developed method is robust and sensitive enough to apply in bioequivalence studies for E2 measurement in human plasma, even at very low concentrations.

17β-雌二醇（E2）是人体中的内源性类固醇。它的测量对于健康和人类生物学理解很重要。然而，人血浆中的E2浓度在pg / mL的范围内，这使其难以检测。这样，尽管E2是一种弱电离的分子，但LC-MS / MS被证明是最灵敏的工具。在这项工作中，我们验证了一种用于人体血浆中E2定量的更灵敏，准确的方法。我们的提取步骤确保了色谱法的更清洁，从而实现了2–150 pg / mL范围内的精确测量和高度可重复的方法。此外，我们证明了E2在多种条件下均具有长期稳定性。所有结果表明，我们开发的方法既稳健又灵敏，足以应用于生物等效性研究中，即使在非常低的浓度下也可用于人体血浆中的E2测量。