Pulsed Electric Field Extracts Obtained from Edible Mushrooms: A Detailed ICP-MS Analysis of Their Mineral and Heavy Metal Contents and Their Cytotoxic Effect on CACO-2 Cells

Mushrooms contain beneficial micro and macronutrients, as well as biologically active compounds, but they can pose a food safety risk by absorbing heavy metals from the environment. Therefore, pulsed electric fields (PEF) technology is of interest as it enables selective extraction of high-added-value compounds, offering an advantage over conventional methods. In this study, PEF extractions were performed under optimal conditions (2.5 kV/cm, 50 kJ/kg) from Agaricus bisporus, Lentinula edodes, Agaricus brunnescens, Pleurotus ostreatus, and Lactarius deliciosus, and compared to aqueous maceration at 20 degrees C. The total carbohydrate content was evaluated by spectrophotometric methods, and the mineral (Ca, Mg, Fe, Zn, P, K, Cu, and Se) and heavy metal (As, Cd, Hg, and Pb) contents were determined by inductively coupled plasma mass spectrometry (ICP-MS). In addition, the contents of the mycotoxins AFB1, AFB2, AFG1, AFG2, OTA, ENNA, EN-NA1, ENNB, and ENNB1 were previously determined by UHPLC-MS/MS. The results showed that PEF extracts achieved higher carbohydrate yields, with increases of up to 1327.02% compared to conventional methods, with L. edodes and P. ostreatus standing out for their content. Different mineral and heavy metal profiles were observed, with L. edodes showing higher levels of As and Cd and A. bisporus of Pb. No mycotoxins were found in the original matrices. Furthermore, PEF extracts (0.78-25% v/v) demonstrated enhanced cytotoxic effects on human colorectal adenocarcinoma cells (CACO-2), suggesting that bioactive compounds, rather than heavy metals, were responsible for reducing cell viability. These findings highlight the PEF technology potential for extracting beneficial compounds from mushrooms, with minimal contamination from heavy metals.