In a subset of 310 participants, we measured baseline urinary metal concentrations including iAs and performed arsenic speciation at the Trace Element Laboratory of the University of Graz, Austria. The participants were chosen from the MESA study using a random site and race/ethnicity stratification to provide predetermined distribution by site and race, resulting in 90 White, 75 Black, 75 Hispanic, and 70 Chinese American participants. Spot urine samples were collected in the morning of the baseline exam in 2000–2002. Urinary arsenic was measured using inductively coupled plasma mass spectrometry (ICPMS, Agilent, Waldbronn, Germany) using standard protocol [43 (link)]. Arsenic speciation was measured using anion-exchange high performance liquid chromatography (HPLC, Agilent, Waldbronn, Germany) coupled to ICPMS [43 (link)]. All arsenic concentrations were corrected for urine dilution using specific gravity [44 (link),45 (link)]. When measurements were below the level of detection (LOD) of 0.10 μg/L, values were replaced with LOD/√2. A total of 133 samples were below the LOD for iAs (46.0%) and 40 samples were below the LOD for MMA (13.8%).
Icp ms
Manufactured by Agilent Technologies
Sourced in United States, Japan, Germany, China
The ICP-MS (Inductively Coupled Plasma Mass Spectrometry) is a laboratory instrument used for the detection and quantification of trace elements in a wide range of sample types. It combines the high-temperature ionization capabilities of an inductively coupled plasma with the high sensitivity and selectivity of a mass spectrometer.
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Suprapur, by Merck Group (3 mentions)
Lsx 213 g2 laser, by Agilent Technologies (3 mentions)
Bca kit, by Thermo Fisher Scientific (3 mentions)
Tris hcl, by Merck Group (2 mentions)
Mili q water, by Merck Group (2 mentions)
1100 hplc, by Agilent Technologies (2 mentions)
1200 infinity lc system, by Agilent Technologies (2 mentions)
Zetasizer nano z, by Malvern Panalytical (2 mentions)
Gc ms, by Agilent Technologies (2 mentions)
Phosphatase inhibitor cocktail set 2, by Merck Group (2 mentions)
Ics 3000, by Thermo Fisher Scientific (2 mentions)
Nitric acid, by Merck Group (2 mentions)
Gemini 2, by Zeiss (1 mentions)
Dsa 100e, by Krüss (1 mentions)
D8 advance, by Bruker (1 mentions)
Tensor 27, by Bruker (1 mentions)
Optical microscopy, by Olympus (1 mentions)
Standard solution, by Merck Group (1 mentions)
0.45 μm membrane, by Cytiva (1 mentions)
Ms excel, by OriginLab (1 mentions)
138 protocols using icp ms
1
Racial Differences in Urinary Arsenic Levels
2
Quantifying Cu2+ in Cell Culture
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Cu2+ concentration in the cell culture medium was quantified by inductively coupled plasma - mass spectroscopy (ICP-MS, Varian, Darmstadt, Germany) against standard solutions of 50 and 100 µg/l.
3
Characterization of Mg Scaffold Biomaterials
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The morphology of the surface and cross section of different scaffolds were observed by a field emission scanning electron microscope (SEM, Gemini 2, Zeiss). The diameter distribution of Mg microspheres was analysed by ImageJ software based on the SEM images. The surface functional groups were identified using a Fourier transform infrared spectrometer (FTIR, TENSOR 27, Bruker). The chemical components of the scaffolds were characterized by TGA (Q500, TA) and XRD (D8 Advance, Bruker). The contact angle was measured by a contact angle meter (DSA100E, KRUSS). The 200 mg scaffold was immersed in 2 ml of deionized water and incubated on a shaker at 80 rpm and 37 ℃. The concentrations of Mg2+ released from the scaffold at 3, 9, 15 and 21 days were measured using inductively coupled plasma–mass spectrometry (ICPMS, Varian, Darmstadt, Germany).
4
Characterizing Ion Release from 3D Scaffolds
5
Cellular Uptake of G4@IONPs
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Cellular uptake of G4@IONPs was evaluated by Prussian blue staining and measured by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) [38 (link),39 (link)]. Different concentrations of G4@IONPs (500, 250, 100, 50, and 0 (control) μg/mL) were added to cell culture media. After 2 h incubation, cells were fixed with 4% formalin and incubated with 4% potassium ferrocyanide and 4% hydrochloric acid (50%, v/v) for 20 min. Finally, the G4@IONPs cellular localization was observed by optical microscopy (Olympus, Tokyo, Japan). Furthermore, MC4L2 cells were trypsinized after 2 h incubation with G4@IONPs (500, 250, 100, 50, and 0 (control) μg/mL) and lysed by 2 mL 65% nitric acid; the quantity of cellular uptake of G4@IONPs was assessed using ICP-MS (Varian Inc, Palo Alto, CA, USA). To obtain the iron concentration per cell, the total iron concentration measured by ICP-MS was divided by the number of lysed cells.
6
Prussian Blue Staining for Iron Detection in MCF7 Cells
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For Prussian blue staining, used to detect the presence of iron, MCF7 cells were incubated in a medium containing G4@IONPs (500 µg/ml) for 2 h. Subsequently, cells were fixed with 4% formalin at room temperature for 20 min and washed with PBS, followed by the incubation with 10% potassium ferrocyanide in 10% HCl (50%, v/v) for 20 min (Samanta et al. 2008 (link)). MCF7 cells after 2 h incubation with 500, 250, 100, 50, and 0 (control) μg/ml G4@IONPs were trypsinized and collected by centrifugation. The collected cells were lysed by 2 ml 65% nitric acid. The amount of the nanoparticles cell uptake was quantified using inductively coupled plasma mass spectrometry (ICP-MS) (Varian Inc, Palo Alto, CA) and the resulting concentration was divided by counting the cell numbers.
7
Characterization of Anaerobic Digestion Byproducts
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Raw slurry and the different fractions obtained
(i.e., S1–S8) were sampled three times over six months of the
observation period (January – Winter, May – Spring,
and July – Summer of 2021). During each sampling event, homogenized
samples (2000 mL each divided in four samples) were collected from
the different separation stages at noon. The samples were collected
in polyethylene sampling containers and transported within 2 h from
the facility to the laboratory in cooler boxes filled with ice. Samples
were stored at 4 °C. Each sample was tested in triplicate.
The following parameters were measured: dry matter at 105 °C
(DM 105 °C), dry matter at 600 °C (DM 600 °C),27 and total Kjeldhal nitrogen (TKN) (EN 13652);28 ammonia-N (NH4-N) (ISO 5664 method);29 nutrients (P, K, Ca, Mg, Fe, Mo, Mn), heavy
metals (Cd, Cr tot, Ni, Pb, Cu, Zn, Hg, Al), and microelements (As,
Co, Se) according to DIN EN ISO 1188530 and UNI-EN 16174.31 Elemental analyses
were carried out by using an inductively coupled plasma mass spectrometry
(ICP-MS, Varian Inc., Fort Collins, CO, USA).
(i.e., S1–S8) were sampled three times over six months of the
observation period (January – Winter, May – Spring,
and July – Summer of 2021). During each sampling event, homogenized
samples (2000 mL each divided in four samples) were collected from
the different separation stages at noon. The samples were collected
in polyethylene sampling containers and transported within 2 h from
the facility to the laboratory in cooler boxes filled with ice. Samples
were stored at 4 °C. Each sample was tested in triplicate.
The following parameters were measured: dry matter at 105 °C
(DM 105 °C), dry matter at 600 °C (DM 600 °C),27 and total Kjeldhal nitrogen (TKN) (EN 13652);28 ammonia-N (NH4-N) (ISO 5664 method);29 nutrients (P, K, Ca, Mg, Fe, Mo, Mn), heavy
metals (Cd, Cr tot, Ni, Pb, Cu, Zn, Hg, Al), and microelements (As,
Co, Se) according to DIN EN ISO 1188530 and UNI-EN 16174.31 Elemental analyses
were carried out by using an inductively coupled plasma mass spectrometry
(ICP-MS, Varian Inc., Fort Collins, CO, USA).
8
Comprehensive Chemical Characterization of Digestate
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Ingestate, digestate and ammonium sulphate, if not better specified in tables, were sampled during a period of 42 months (from January 2017 to June 2020) and characterized from a chemical point of view according, also, (digestate) to Lombardy Region regulation (Regione Lombardia, 2019) for agricultural use of these products.
The following parameters were determined: pH (EPA 9045D) (EPA, 2004) , dry matter at 105°C (DM 105°C), dry matter at 600°C (DM 600°C) and Total Organic Carbon (TOC) (APHA, 1998), total nitrogen (TKN) (EN 13652) (EN 2001) . Ammonia-N (NH4-N) was determined by ISO 5664 method (ISO, 1984) (reagent and grade in Table S1).
In addition, nutrients (P, K, Ca, Mg, Fe, Mo, Mn), heavy metals contents (Cd, Cr tot, Ni, Pb, Cu, Zn), and micropollutants/element (Hg, As, Al, Co, Se, Na) were determined by inductively coupled plasma mass spectrometry (ICP-MS, Varian Inc., Fort Collins, CO, USA) according to DIN EN ISO 11885 (ISO, 2009) (Ca, Mn, Mg, Fe, Mo, Al, Co, Na) , and according to UNI-EN 16174 (UNI-EN, 2012) for all the other elements.
The following parameters were determined: pH (EPA 9045D) (EPA, 2004) , dry matter at 105°C (DM 105°C), dry matter at 600°C (DM 600°C) and Total Organic Carbon (TOC) (APHA, 1998), total nitrogen (TKN) (EN 13652) (EN 2001) . Ammonia-N (NH4-N) was determined by ISO 5664 method (ISO, 1984) (reagent and grade in Table S1).
In addition, nutrients (P, K, Ca, Mg, Fe, Mo, Mn), heavy metals contents (Cd, Cr tot, Ni, Pb, Cu, Zn), and micropollutants/element (Hg, As, Al, Co, Se, Na) were determined by inductively coupled plasma mass spectrometry (ICP-MS, Varian Inc., Fort Collins, CO, USA) according to DIN EN ISO 11885 (ISO, 2009) (Ca, Mn, Mg, Fe, Mo, Al, Co, Na) , and according to UNI-EN 16174 (UNI-EN, 2012) for all the other elements.
9
Characterization of Anaerobic Digestion Plants
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Ingestates, digestates, liquid fractions of digestates and solid fractions of digestates were sampled from six mesophilic full-scale anaerobic digestion plants located in the Lombardy Region (North Italy). Plants were characterized by different hydraulic retention times (HRT) and feed-stock mixtures (Table 1). The samples taken were stored in 2 litre PTFE bottles without headspace before chemical characterization.
On fresh samples, the Total N-Kjeldahl (TKN) and ammonia (TAN) were analysed according to the analytical method established for wastewater sludge (APHA 1998) as well as pH (US Department of Agriculture and US Composting Council, 2002) . Total solids (TS) and volatile solids (VS) were determined following standard procedures (APHA, 1998) . On dry samples, total P and K contents were quantified, after acid digestion (EPA, 1998) , by inductively coupled plasma mass spectrometry (ICP-MS by Varian, Fort Collins, USA); standard samples (National Institute of Standards and Technology, Gaithersburg, MD, USA) and blanks were run with all samples to ensure precision in the analyses. All the sampling and the analyses were carried out in triplicate.
On fresh samples, the Total N-Kjeldahl (TKN) and ammonia (TAN) were analysed according to the analytical method established for wastewater sludge (APHA 1998) as well as pH (US Department of Agriculture and US Composting Council, 2002) . Total solids (TS) and volatile solids (VS) were determined following standard procedures (APHA, 1998) . On dry samples, total P and K contents were quantified, after acid digestion (EPA, 1998) , by inductively coupled plasma mass spectrometry (ICP-MS by Varian, Fort Collins, USA); standard samples (National Institute of Standards and Technology, Gaithersburg, MD, USA) and blanks were run with all samples to ensure precision in the analyses. All the sampling and the analyses were carried out in triplicate.
10
Quantitative Ion Concentration Analysis
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Ion concentrations (Ca2+, P as PO43-, Cr3+) in the cell culture supernatants and in cement extracts were determined using inductively-coupled-plasma mass-spectrometry (ICP-MS, Varian, Germany). The quantitative measurement was carried out against standard solutions (Merck) containing defined concentrations of all ions of interest.
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