Roots and shoots were harvested separately. Roots were first washed 10 min with 2 mM CaSO4 and 10 mM EDTA, then washed 3 min with 0.3 mM bathophenanthroline disulphonate and 5.7 mM sodium dithionite and finally rinsed in deionized water. Shoots were rinsed with deionized water. Samples were dried at 80 °C for at least 2 days and mineralized in 7.5% H2O2, 49% HNO3 at 120 °C. Elemental analyses were performed by Micro Plasma Atomic Emission Spectroscopy (Agilent 4200 MP-AES) according to the manufacturer’s recommendations.
Agilent 4200 mp aes
Manufactured by Agilent Technologies
Sourced in United States, Australia
The Agilent 4200 MP-AES is a microwave plasma atomic emission spectrometer. It utilizes a microwave-induced plasma to atomize and excite the sample, and then measures the characteristic emission spectra of the elements present in the sample to determine their concentrations.
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Lab products found in correlation
Cm30 tem stem, by Philips (1 mentions)
Syringe filter, by Sartorius (1 mentions)
Multi element standard solution, by Merck Group (1 mentions)
0.2 m syringe filter, by Sartorius (1 mentions)
Liquid chromatograph mass spectrometer, by Shimadzu (1 mentions)
Complete mini, by Roche (1 mentions)
Mc03f, by Bangs Laboratories (1 mentions)
Pkh26, by Merck Group (1 mentions)
22 protocols using agilent 4200 mp aes
1
Elemental Analysis of Plant Samples
2
Characterization of Magnetic Nanoparticles
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Concerning their physicochemical features, MNPs were analyzing regarding their size, iron content according to previous studies (Friedrich et al. 2021 ; Mühlberger et al. 2019 (link); Stein et al. 2020 ). Their iron content in milligram per milliliter was investigated after diluting them 1:20 in deionized H2O, liquefying them in 65% nitric acid with atomic emission spectroscopy (AES), using Agilent 4200 MP-AES with an iron solution of 1000 mg Fe/l as an external standard (Bernd Kraft, Duisburg, Germany). MNP particles were diluted to the desired concentration with deionized water for all experiments. Transmission electron microscopy (TEM) images were acquired using the CM30 TEM/STEM (Philips, The Netherlands) operating at 300 kV acquired with a CCD camera, a Tietz Fast Scan-F114 (Tietz Video and Image Processing Systems GmbH, Gauting, Germany). Dispersions at a concentration of 100 µg Fe/ml were dropped onto a carbon-coated copper grids (Plano, Germany) an air-dried at RT.
3
Determination of Iron by AES
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For the determination of the iron concentration, atomic emission spectroscopy (AES) was used (Agilent 4200 MP-AES, Agilent Technologies, Santa Clara, CA, USA). A commercially available iron solution (1.000 mg/L, Bernd Kraft, Duisburg, Germany) served as an external standard. Samples were diluted at a ratio of 1:20. To 20 μL of the diluted sample, 80 μL of 65% nitric acid was added. The mixture was heated to 95 °C for 10 min and afterward diluted with water to 2 mL. Triplicate measurements were carried out at a wavelength of 371.993 nm and the results were averaged.
4
Synthesis and Characterization of SPIONs
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SPIONs were synthesized based on an adjusted protocol of Elbialy et al. [37 (link)] in three batches. Particles were sterilized by filtration through syringe filters with 0.2 µm pore size (Sartorius, Goettingen, Germany). Subsequently, SPIONs were analyzed regarding their size, iron content, magnetic susceptibility, and zeta potential according to Mühlberger et al. [26 (link)]. The iron content was investigated after a dilution of 1:25 in deionized H2O and dissolving in 65 % nitric acid with atomic emission spectroscopy (AES), using the Agilent 4200 MP-AES (Agilent Technologies, Santa Clara, CA, USA) with an iron solution of 1000 mg/l as an external standard (Bernd Kraft, Duisburg, Germany). Triplicate measurements were performed at a wavelength of 371,993 nm, which were then averaged.
5
Quantitative Mineral Analysis Protocol
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The contents of K, Na, Ca, Mg, Fe, Cu, Mn, and Zn were determined by atomic absorption spectroscopy following wet mineralization, and using the instrumental condition as previously described [28 (link)]. Briefly, the samples were digested, and approximately 100 mg of dried sample was weighed and incubated with 9 mL of 65% (w/w) HNO3, and 1 mL of 30% (w/v) H2O2 were added. The temperature was set at 200 °C for 20 min. Once cooled, the digested samples were diluted to a final volume of 50 mL with distilled H2O. All measurements were performed using an Agilent 4200 MP-AES fitted with a double-pass cyclonic spray chamber and OneNeb nebulizer. The calibration standards were prepared by diluting a 1000 mg/L multi-element standard solution (Sigma Aldrich and Scharlab S.L.) in 1% (v/v) HNO3. Finally, P was determined using a colorimetric method [29 ]. Data were expressed as mg per 100 g of Fresh Weight (PW).
6
Mineral Content Analysis of Freeze-Dried Extracts
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Freeze-dried pooled extracts were digested in a combination of nitric acid (HNO3) and hydrogen peroxide on a hot plate and evaporated until dryness (up to 24 h). Digested samples were diluted in 20 mL of 5% HNO3 and analysed for mineral content by Microwave Plasma-Atomic Emission Spectrometer (MP-AES; Agilent 4200 MP-AES, Agilent Victoria, Australia), as described in Pereira et al.6 (link). Instrumental detection limits were as follows: Ca: 0.04 μg/L, Cd: 1.4 μg/L, Cr: 0.3 μg/L, Cu: 0.5 μg/L, Fe: 1.7 μg/L, K: 0.6 μg/L, Mg: 0.031 mg/L, Mn: 0.1 μg/L, Na: 0.1 μg/L, Ni: 1.1 μg/L, Pb: 2.5 μg/L and Zn: 3.1 μg/L. Results were expressed as mg or μg/g of extract dry weight (DW). Appropriate blanks were also produced and analysed.
7
Synthesis and Characterization of SPIONs
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SPIONs were synthesized as described previously using an adjusted protocol by Elbialy et al. (23 (link), 32 (link)) The nanoparticles were sterilized by filtration through a 0.2 µm syringe filter (Sartorius, Goettingen, Germany). The hydrodynamic size, iron concentration, magnetic susceptibility, and zeta potential of the SPIONs were then characterized as described by Mühlberger et al. and Boosz et al. (21 (link), 23 (link)) The iron content was investigated at a dilution of 1:25 in dH2O, dissolved in 65% nitric acid, using atomic emission spectroscopy with an Agilent 4200 MP-AES (Agilent Technologies, Santa Clara, CA, USA) with an iron solution of 1000 mg Fe/L as an external standard (Bernd Kraft, Duisburg, Germany). Measurements were performed in triplicates at a wavelength of 371.993 nm, which were then averaged. SPIONs were diluted with sterile dH2O to the intended concentration for all experiments.
8
Quantifying Nanoparticle Biodistribution in Mice
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All animal studies were approved by the Ethical Committee of Pirogov Russian State Medical University (protocol ## 25/2017, 26/2017). Eight-to-ten-week-old female BALB/c mice bearing CT26 tumors received MNP-HSA@PS by single tail vein injection at dose of 5 mg iron/kg. Animals were sacrificed at 1, 4, and 24 h after i.v. injection (n = 4 for each time point) and tumor, liver, spleen, and kidney were collected and dissolved in concentrated nitric acid (48 h incubation at room temperature). Iron concentration in the tissue samples was measured by inductively coupled plasma-atomic emission spectrometry (Agilent 4200 MP-AES, Santa Clara, CA, USA). Untreated mice (n = 4) were used as control to identify endogenous (background) iron concentration.
9
Liver Iron Content and Redox Quantification
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For hepatic iron content, approximately 30 mg of liver tissue was homogenized by sonication (Fisher Scientific Sonic Dismembrator F60, Fair Lawn, NJ, USA) in 300 µL of RIPA buffer (Roche Complete Mini #11836153001, Roche, UK). After increasing the volume to 600 µL with distilled water, 150 µL of 40% nitric acid (Fisher Scientific metal free grade, CAS #7697-37-2) was added. Samples were digested for 90 min at 95 °C and then centrifuged at 12,000 rpm for 20 min. Supernatants were diluted to a volume of 4 mL using 1% nitric acid and filtered through 0.45 µm nylon syringe filters (Fisher Scientific #09-719-008), prior to loading for run in the MIP OES (Microwave-Induced Plasma Optical Emission Spectrometer, Agilent 4200 MP-AES, Santa Clara, CA, USA). To measure redox state in the livers, reduced glutathione (GSH) and glutathione disulfide (GSSG) were quantified using LC-MS/MS (Liquid Chromatograph Mass Spectrometer, Shimadzu 8050, Japan).
10
Dissolution Kinetics of Cu2O Particles
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