Nexion 300d

All waste materials were spiked with the acid mine drainage (AMD) prepared from the oxidation of pyritic tailings. This experience was made by the addition of 50 mL of AMD to 10 g of each waste material in triplicate to check the first impact of the AMD on different waste materials. Afterward, they were stirred for 24 h and filtered (Filter-Lab n°1250, pore size: 10–13 µm), separating the waste material (solid phase) from the leachate (liquid phase). In the leachate, which is the AMD treated, pH_(L) and EC_(L) were measured with a pH/conductivity-meter 914 Metrohm (Metrohm AG, Herisau, Switzerland) and an Eutech CON700 conductivity-meter (Oakton Instruments, Vernon-Hills, IL, Waltham, MA, USA), respectively, and PTE concentrations in solution were determined by inductively coupled plasma optical mass spectrometry (ICP-MS) in a spectrometer PerkinElmer NexION 300D (PerkinElmer, Inc., Waltham, MA, USA). The precision and accuracy of this method were assessed by measurement (three replicates) of a certified reference material (CRM BCR–482 EC-JRC-IRMM, Geel, Belgium). For all elements of interest, measured values were within the prediction interval of the certified value.

The cells were digested with 2% nitric acid solution (Sigma-Aldrich) for 20 min and centrifuged. The cell debris was discarded, and internal standard (yttrium) was added. The boron (¹⁰B) concentration in the supernatant was measured using ICP-MS (PerkinElmer NexION 2000B, PerkinElmer Inc., Waltham, MA, USA) with high-purity argon. The blood, skin, and tumor were digested with 70% nitric acid solution at 100 °C for 1 h in Thermomixer (Eppendorf, Hamburg, Germany) and dilution with water and internal standard (yttrium). The boron (¹⁰B) concentration was measured using ICP-MS; (PerkinElmer NexION 300D, PerkinElmer Inc., USA) with high-purity argon. Confirmation of the reliability of the measurement method was confirmed using six concentrations prepared using the standard solution.

An inductively coupled plasma mass spectrometer, ICP-MS, NexION 300D (Perkin Elmer, Boston, MA, USA), equipped with a quartz cyclonic spray chamber and Meinhard nebuliser, was used for Al and Au concentration measurements.

A NexIon 300 D (Perkin Elmer, Sciex, Toronto, Canada) was operated as ICP-MS system for the on-line detection of CE-efflux. The isotopes ⁵⁶Fe and ⁵⁷Fe were measured in dynamic reaction cell (DRC) mode. The RF power was set to 1,250 W, the plasma gas was 16 L Ar/min. The nebulizer gas was optimized and finally set to 0.98 L Ar/min. The dwell time was 50 ms. Ammonia was used as DRC gas (0.58 ml NH₃/min) and DRC rejection parameter was set to 0.58.

AuNP stock solution was used to prepare the calibration solutions through serial aqueous dilutions for ICP-MS determination (NexION 300D, PerkinElmer). 1% (v/v) nitric acid (Merck) and 10 μg/L rhodium from Perkin-Elmer was added to the calibration solution, blank, and samples to improve analytical performance. The Au¹⁹⁷ isotope was measured. For quantification of intracellular Au mass, spheroids of both models were treated for 24 or 72 h with Au-SC or Au-PEG. Then, spheroids were collected and lysed (CelLytic M lysis buffer, Molecular Probes) for analysis.

The elemental concentrations in different tissues of rice plants or yeast cells were determined by an inductively coupled plasma mass spectrometer (ICP-MS) (NexION 300D; PerkinElmer Corp., Norwalk, CT, United States) according to previous studies (Huang et al., 2016 (link), 2019 (link)). The roots and different leaves of WT, osmot1;2, and OsMOT1;2 overexpression lines grown hydroponically and different tissues at mature growth stages were harvested as described earlier (Huang et al., 2019 (link)).

Five-day-old seedlings of WT, rae2, and rae3/hpr1 grown on 0.4% gellan gum (G1910, Sigma-Aldrich) medium containing the full-strength Hoagland nutrient and 1% sucrose were pretreated with a 2% MGRL nutrient solution containing 1% sucrose at pH 4.8 for 2 h and then exposed to the same solution containing 0 or 20 μM Al at pH 4.8 for 12 h. Root exudates were collected for malate measurement by the NAD/NADH enzymatic cycling method (Hampp et al., 1984 (link)). For the determination of Al content, three-week-old seedlings of WT and rae2 grown on the one-tenth-strength Hoagland solution were pretreated with 0.5 mM CaCl₂ solution for 6 h at pH 4.8 and then treated with 0.5 mM CaCl₂ solution containing 0 or 30 μM Al for 12 h at pH 4.8. Al content in roots was determined according to previous methods (Ligaba-Osena et al., 2017 (link); Zhang et al., 2019 (link)). The Al concentration in a diluted solution was measured by inductively coupled plasma mass spectrometry (ICP-MS; PerkinElmer NexION300D).