All digested samples were analyzed for total metals concentrations using the Agilent 7900 ICP-MS (Agilent Technologies, Santa Clara, CA) with or without helium collision mode depending on the element in accordance with EPA Method 6020B. The instrument was tuned using a 10-ppb solution of Ce, Co, Li, Tl, and Y in 5% nitric acid (Inorganic Ventures, Christiansburg, VA) and a 1-ppm solution of Li, Sc, Ge, Y, In, Tb, and Bi in 5% nitric acid (Inorganic Ventures) was used as an internal standard. The glassware used consisted of 0.4 mL/min flow concentric glass MicroMist™ nebulizer, quartz Scott-style spray chamber, straight quartz connector, and quartz torch with 2.5 mm injector (Glass Expansion, Pocasset, MA). Instrument settings consisted of 1500 W RF power, 15.00 L/min plasma gas flow rate, 1.1 L/min carrier gas flow rate, 0.9 L/min auxiliary gas flow rate, and 2 °C spray chamber temperature. Method detection limits (MDLs) and method reporting limits (MRLs) were determined separately for each digestion method according to 40 CFR Appendix B to Part 136 and are provided in Table S1 in the supporting information . In the results for all samples, the values from individual replicates were either presented as measured when greater than the MDL or as one-half of the MDL when less than the MDL, as long as the average of the replicates was greater than the MDL.
Nitric acid
Manufactured by Inorganic Ventures
Nitric acid is a strong, corrosive acid composed of nitrogen, hydrogen, and oxygen. It is a clear, colorless to yellow liquid with a pungent, acrid odor. Nitric acid is commonly used in various industrial and laboratory applications that require a strong oxidizing agent.
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3 protocols using nitric acid
1
Analysis of Trace Metal Concentrations
2
Analysis of Trace Metal Concentrations
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All digested samples were analyzed for total metals concentrations using the Agilent 7900 ICP-MS (Agilent Technologies, Santa Clara, CA) with or without helium collision mode depending on the element in accordance with EPA Method 6020B. The instrument was tuned using a 10-ppb solution of Ce, Co, Li, Tl, and Y in 5% nitric acid (Inorganic Ventures, Christiansburg, VA) and a 1-ppm solution of Li, Sc, Ge, Y, In, Tb, and Bi in 5% nitric acid (Inorganic Ventures) was used as an internal standard. The glassware used consisted of 0.4 mL/min flow concentric glass MicroMist™ nebulizer, quartz Scott-style spray chamber, straight quartz connector, and quartz torch with 2.5 mm injector (Glass Expansion, Pocasset, MA). Instrument settings consisted of 1500 W RF power, 15.00 L/min plasma gas flow rate, 1.1 L/min carrier gas flow rate, 0.9 L/min auxiliary gas flow rate, and 2 °C spray chamber temperature. Method detection limits (MDLs) and method reporting limits (MRLs) were determined separately for each digestion method according to 40 CFR Appendix B to Part 136 and are provided in Table S1 in the supporting information . In the results for all samples, the values from individual replicates were either presented as measured when greater than the MDL or as one-half of the MDL when less than the MDL, as long as the average of the replicates was greater than the MDL.
3
Quantitative Fe Analysis by ICP-MS
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ICP-MS experiments were performed at the Quantitative Bio-element Imaging Center at Northwestern University. The samples were digested in concentrated nitric acid (Thermo Fisher Scientific) by incubating at 65 °C for 1 h and were then diluted with Milli-Q water to a final solution of 3% nitric acid. An element standard containing Fe (Inorganic Ventures) was used to prepare a 100-ng/mL standard solution in 3% nitric acid (vol/vol). Online dilution was carried out by the prepFAST system and was used to generate calibration curves consisting of 100, 50, 10, 2, 1, and 0.5 ng/g of Fe. Samples were measured using a Thermo iCAPQ ICP-MS instrument equipped with a CETAC ASX260 autosampler (Thermo Fisher Scientific). The isotopes selected for analysis were 56Fe, 57Fe, 89Y, and 115In (89Y and 115In were used as internal standards for data interpolation and machine stability). The data were analyzed using QTEGRA software. All reported values were measured with three to five independent protein samples.
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