Icap qc quadrupole icp mass spectrometer

To measure the cellular uptake of 1–3, about 1 million HMLER-shEcad cells were treated with 1–3 (5 μM) at 37 °C for 24 h. After incubation, the media were removed, and the cells were washed with PBS (2 mL × 3), and harvested. The number of cells was counted at this stage, using a haemocytometer. This mitigates any cell death induced by 1–3 at the administered concentration, and experimental cell loss. The cellular pellet was dissolved in 65% HNO₃ (250 µL) overnight. All samples were diluted 17-fold with water, and analysed using inductively coupled plasma mass spectrometry (ICP-MS, using the Thermo Scientific ICAP-Qc quadrupole ICP mass spectrometer). Copper levels are expressed as mass of Cu (ng) per million cells. Results are presented as the mean of four determinations for each data point.

To measure the cellular uptake of 1 and 3, about 1 million HMLER‐shEcad cells were treated with 1 or 3 (at 5 μm) at 37 °C for 24 h. After incubation, the media was removed, the cells were washed with PBS (2 mL × 3), and harvested. The number of cells was counted at this stage, using a haemocytometer. This mitigates any cell death induced by 1 and 3 at the administered concentration and experimental cell loss. The cellular pellets were dissolved in 65 % HNO₃ (250 μL) overnight. Cellular pellets of 1 and 3 treated HMLER‐shEcad cells were also used to determine the nickel content in the nuclear, cytoplasmic, and membrane fractions. The Thermo Scientific NE‐PER Nuclear and Cytoplasmic Extraction Kit was used to extract and separate the nuclear, cytoplasmic, and membrane fractions. The fractions were dissolved in 65 % HNO₃ overnight (250 μL final volume). All samples were diluted fivefold with water and analysed using inductively coupled plasma mass spectrometry (ICP‐MS, ThermoScientific ICAP‐Qc quadrupole ICP mass spectrometer). Nickel levels are expressed as Ni (ppb) per million cells. Results are presented as the mean of four determinations for each data point.

All synthetic procedures were performed under normal atmospheric conditions. ¹H and ³¹P{¹H} NMR were recorded at room temperature on a Bruker Avance 400 spectrometer (¹H 400.0 MHz, ³¹P 162.0 MHz) with chemical shifts (δ, ppm) reported relative to the solvent peaks of the deuterated solvent. Fourier transform infrared (FTIR) spectra were recorded with an IRAffinity-1S Shimadzu spectrophotometer. UV–Vis absorption spectra were recorded on a Cary 3500 UV–Vis spectrophotometer. ICP-MS was performed using a Thermo Scientific, Waltham, MA, USA, ICAP-Qc quadrupole ICP mass spectrometer. Elemental analysis of the compounds prepared was performed commercially by the University of Cambridge. Cu(NO₃)(PPh₃)₂ was prepared, using a previously reported protocol [32 ]. Sodium salts of the NSAIDs (naproxen, diclofenac, and salicylic acid) were purchased from Sigma-Aldrich, Burlingthon, MA, USA, and used without further purification. Solvents were purchased from Fisher, and used without further purification.