PBCECs were cultivated in rat tail collagen-coated 24 well culture plates and cultured according to the cultivation in Transwell® filters. After 72 h incubation of the respective Hg species, PBCECs were washed twice with PBS (100 mM NaCl, 4.5 mM KCl, 7 mM Na2HPO4, 3 mM KH2PO4 (all Sigma Aldrich); pH 7.4) and incubated with 120 μL lysis buffer (RIPA-buffer; 0.01 M Tris, pH 7.6, 0.15 M NaCl, 0.001 M EDTA, 1% sodium desoxycholate, 0.1% (all Sigma Aldrich)) for 15 min on ice. After scrapping off and sonication, the suspension of lysed cells was centrifuged at 10 000 x g for 20 min at 4 °C. Total cellular Hg content was quantified by inductively coupled mass spectrometry (ICP-MS; Agilent 8800 ICP-QQQ, Agilent Technologies Deutschland GmbH, Boeblingen, Germany) in an aliquot of the supernatant. The Bradford assay was used to determine the cellular protein level.
Agilent 8800 icp qqq
Manufactured by Agilent Technologies
Sourced in Germany, United States
The Agilent 8800 ICP-QQQ is a triple quadrupole inductively coupled plasma mass spectrometer (ICP-QQQ) designed for trace elemental analysis. It utilizes two quadrupole mass filters in series to provide enhanced selectivity and sensitivity for the detection of analytes. The core function of the Agilent 8800 ICP-QQQ is to perform precise and accurate multi-element analysis of a wide range of sample types.
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Lab products found in correlation
Sodium desoxycholate, by Merck Group (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Kh2po4, by Merck Group (1 mentions)
Na2hpo4, by Merck Group (1 mentions)
Cm200 electron microscope, by Philips (1 mentions)
Unicam uv500 spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Micromist nebulizer, by Agilent Technologies (1 mentions)
Nano zs90, by Malvern Panalytical (1 mentions)
Varian cary eclipse fluorescence spectrometer, by Agilent Technologies (1 mentions)
10 protocols using agilent 8800 icp qqq
1
Quantifying Cellular Mercury Content
2
Maternal RBC Manganese Quantification
3
Synthesis and Characterization of AgNPs
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The working pH was adjusted using a pH meter GLP 21 Crison (Barcelona, Spain). A UNIVERSAL 320 centrifuge (Hettich-Tuttlingen, Germany) was used in the synthesis of silver nanoparticles stabilized with PVP. Absorption spectra of AgNPs were obtained with a Thermo Spectronic UNICAM UV 500 spectrophotometer (Loughborough, UK) equipped with plastic disposable cells with optic path of 10 mm and spectral resolution of 1 nm in the range of 220–800 nm. Transmission electron microscopy (TEM) measurements were recorded with a Phillips CM 200 electron microscope (Amsterdam, the Netherlands) working at 200 KV. An Inductive Coupled Plasma Mass Spectrometer (ICP-MS) (Agilent 8800 ICP-QQQ, Agilent Technologies, Santa Clara, CA, USA) was used for aluminum content measurements in water samples.
4
Toenail Metal Exposure Assessment
5
Uranium Isotopes Separation and Purification
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The separation and purification procedure for U isotopes was conducted following the method of Yang et al.27 (link). About 1 g soil samples were ashed in a muffle oven at 450 °C for 2 h to decompose organic matter. Total dissolution (HF + HNO3 + HClO4) was performed in PFA jars with lids (Savillex, Eden Prairie, MN, USA) on a hot plate at 180 °C for 1 d. After filtration, Si was removed by reaction with 46% HF. Then, the HF solution was heated to dryness, after which 5 mL of 61% HNO3 was added and this acid solution was heated to dryness to remove residual HF. Subsequently, the sample residue was dissolved into 10 mL of 6 M HNO3, ready for chromatographic purification using DGA resin. After removing interfering elements by 6 M HNO3 and 8 M HNO3, U was eluted from the resin by 15 mL of 0.1 M HNO3. Finally, the U eluate was evaporated to near dryness and dissolved into 1.5 mL of 4% HNO3. A 20 μL aliquot was taken out and diluted with 4% HNO3 at a dilution factor of 2000 for the 238U concentration (activity) measurement via an Agilent 8800 ICP-QQQ operated in the single MS mode (Agilent Technologies, Santa Clara, CA, USA). The remaining portion was analyzed for 234U/238U, 235U/238U, and 236U/238U atom ratios, via the Agilent 8800 ICP-QQQ MS/MS mode operation. Finally, 234U, 235U, 236U activities could be calculated by combining the data of these two mode analyses.
6
Quantifying Mn Concentration in Striatum
7
Quantifying Metal Concentrations in Brain Regions
8
Trace Element Assessment in Toenails
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We used inductively coupled plasma mass spectrometry (ICP-MS) to assess baseline toenail concentrations for a panel of 16 trace elements (antimony, arsenic, cadmium, chromium, cobalt, copper, iron, lead, mercury, manganese, molybdenum, nickel, selenium, tin, vanadium, and zinc). As previously described,15 (link),19 (link) the two big toenails were washed, air-dried, and then digested in acid (9:1 HNO3/HCL) and diluted with deionized water. ICP-MS analyses were conducted using Agilent 8800 ICP-QQQ (Santa Clara, CA). Quality control was conducted using continuous calibration verification, duplicate and spiked samples, within- and between-batch analyses of a laboratory-prepared toenail matrix digest, and comparison with a standard reference material (Japan NIES #13 Hair). Random effect models were used to correct for sample batch. If a measured value was available despite being below the limit of quantification that value was used in the analysis. After batch correction, a small number of metal measurements (<0.1%) were negative. For some calculations that required positive values (e.g., log transformations), these values were imputed as a small positive value (0.001) but were otherwise left negative.
9
Cellular Arsenic Bioavailability and Distribution
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Cellular bioavailability and distribution were studied as described before.14 (link) Briefly, after 48 h of incubation trypsinised cells were pelletised and for total arsenic measurement wet-ashed (acid digestion with HNO3/H2O2 solution (1/1, v/v) at 95 °C for at least 12 h). The arsenic content of the digest was determined by ICP-MS/MS (Agilent 8800 ICP-QQQ, Agilent Technologies, Germany) in the mass-shift mode using oxygen as the reaction gas. ICP-MS/MS parameters are listed in Table 1 . According to the calculated LOQ a cellular arsenic concentration of at least 0.08 μM is quantifiable using the described method when 380 000 cells are seeded.
For distribution analysis, cell pellets were lysed by addition of bi-distilled water and sonication (15 s, 100%, 0.8 cycles). The cytosol was separated from the cell-debris-associated parts by centrifugation (5 min, 23 600g, 4 °C) and the total arsenic content of both fractions was determined by ICP-MS/MS as described above.
For distribution analysis, cell pellets were lysed by addition of bi-distilled water and sonication (15 s, 100%, 0.8 cycles). The cytosol was separated from the cell-debris-associated parts by centrifugation (5 min, 23 600g, 4 °C) and the total arsenic content of both fractions was determined by ICP-MS/MS as described above.
10
Characterization of Photoluminescent Quantum Dots
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QDs photoluminescent spectra were performed with the Varian Cary Eclipse Fluorescence Spectrometer (Agilent, Santa Clara, CA, USA) equipped with a xenon discharge lamp (peak power equivalent to 75 kW), a Czerny-Turner monochromator and photomultiplier tube detector (Model R-298). The emission spectra (1 nm data interval), which presented a maximum emission wavelength at 590 nm, were recorded upon excitation at 290 nm, with a delay time of 0.2 ms and a gate time of 5 ms. Excitation and emission slits were set at 10/10 nm respectively, and the averaging time selected to perform the experiments was 5 ms. A microplate reader accessory was used for phosphorescent immunoassay measurements.
QDs characterization was carried out by simultaneous detection and quantification of the elements constituting the QD core (S, Zn and Mn) using an ICP-MS/MS system (Agilent 8800 ICPQQQ, Tokyo, Japan). For separation and characterization of nanoparticles and their bioconjugates the ICP-MS/MS was coupled on-line to the AF4 system (AF2000, Postnova Analytics, Landsberg, Germany). Separation conditions are summarized in Table S1 (Electronic Supplementary Material, ESM). Dynamic light scattering spectra were measured by using a NanoZS90 instrument from Malvern Instruments, Houston, TX, USA.
QDs characterization was carried out by simultaneous detection and quantification of the elements constituting the QD core (S, Zn and Mn) using an ICP-MS/MS system (Agilent 8800 ICPQQQ, Tokyo, Japan). For separation and characterization of nanoparticles and their bioconjugates the ICP-MS/MS was coupled on-line to the AF4 system (AF2000, Postnova Analytics, Landsberg, Germany). Separation conditions are summarized in Table S1 (Electronic Supplementary Material, ESM). Dynamic light scattering spectra were measured by using a NanoZS90 instrument from Malvern Instruments, Houston, TX, USA.
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