The cytosolic fractions (250 μL) of microvessels, as well as peripheral tissue samples (small intestines, liver and kidneys) isolated from Cu(ATSM) (30 mg/kg) and vehicle-treated mice (n = 6–8) were lyophilised prior to sample preparation for ICP-MS. The wet weights of organs were determined prior to lyophilisation. Small intestines were approximately 150 mg, livers were approximately 150 mg, and the two kidneys had a combined weight of approximately 250 mg. Nitric acid (HNO3) (65% Suprapur, Merck, Darmstadt, Germany) was added to each sample and digested overnight. The samples were further digested at 90 °C for 20 min in a heating block and then an equivalent volume of hydrogen peroxide 30% (v/v) (Aristar, BDH) was added to each sample, which was allowed to digest for 30 min. Samples were allowed to stop effervescing for 30 min before being heated again for 15 min at 70 °C. The average reduced volume was determined, and 50 μL aliquots of samples were further diluted 1:20 for MEF cytosolic fractions and 1:400 for tissue samples with 1% HNO3 (v/v). The ICP-MS measurements were made using an Agilent 7700 series ICP-MS instrument (Santa Clara, CA, USA) with routine multi-element operating conditions using a Helium Reaction Gas Cell. The instrument was calibrated using 0, 5, 10, 50, 100 and 500 ppb of certified multi-element ICP-MS standard calibration solutions (ICP-MS-CAL2-1, ICP-MS-CAL-3 and ICP-MS-CAL-4, Accustandard, New Haven, CT, USA) for a range of elements. Additionally, a certified standard solution containing 200 ppb of Yttrium (Y89) was used as an internal control (ICP-MS-IS-MIX1-1, Accustandard). The elements assessed included sodium, magnesium, phosphorous, calcium, iron, cobalt, nickel, copper, zinc, selenium, and rubidium, and elements with abundances at or below detection limits were excluded from the data analysis.
Agilent 7700 series icp ms instrument
Manufactured by Agilent Technologies
7 citations
Sourced in United States
About the product
The Agilent 7700 series ICP-MS instrument is a versatile and high-performance inductively coupled plasma mass spectrometer designed for elemental analysis. It offers precise and accurate detection of trace elements in a wide range of sample types.
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7 protocols using «agilent 7700 series icp ms instrument»
1
Trace Element Analysis in Mice Tissues
Corresponding organizations : Monash University, University of Melbourne, Florey Institute of Neuroscience and Mental Health
2
ICP-MS Analysis of Metal Content
ICP-MS was performed as previously described [56 (link)]. Briefly, ipsilateral and contralateral cortical brain homogenates were lyophilized and the dry material was digested with 50 µL of Nitric Acid (HNO3, 65% Suprapur, Merck) overnight. The samples were further digested by heating to 90°C for 20 min using a heating block. Samples were removed from the heating block and an equivalent volume of 50 µL of Hydrogen Peroxide (H2O2, 30% Aristar, BDH) was added to each sample. Samples were allowed to stop digesting for ~ 30 min before heating again for 15 min at 70°C. The average reduced volume was determined and the samples further diluted with 1% HNO3. All measurements were performed on an Agilent 7700 Series ICP-MS instrument (Agilent Technologies, CA, USA) under routine multi-element operating conditions using a Helium Reaction Gas Cell. The instrument was calibrated with 0, 5, 10, 50, 100, and 500 ppb of certified multi-element ICP-MS standard calibration solutions (ICP-MS-CAL2-1, ICP-MS-CAL-3 and ICP-MS-CAL-4, Accustandard, CT, USA) for a range of elements. A certified internal standard solution containing 200 ppb of Yttrium (Y89) was used as the internal control (ICP-MS-IS-MIX1-1, Accustandard). Metal content (µg/g) was normalised to protein concentration which was determined using the BCA Protein Assay Kit (Thermo Fisher Scientific).
Corresponding organizations : University of Melbourne, Florey Institute of Neuroscience and Mental Health
3
Quantification of Intracellular Metal Levels
hCMEC/D3 cells were seeded in 24‐well plates using the protocol for cell seeding described above. For every treatment replicate, it was necessary to have one blank replicate that contained no cells. Blanks were treated identically throughout the process (lysed the same way, as though they had cells in them), and were used to blank correct for background metal detection. Following treatment, cells were washed twice with 500 μl of ice‐cold PBS, then lysed with 250 μl of ice‐cold 1% (v/v) Triton X‐100 in MQ water and put on a shaker at 4°C for 15 min. Following lysis, the lysate was homogenised and 240 μl was transferred to pre‐cooled Eppendorf tubes. For blank‐treated wells, only 210 μl of the lysis solution was transferred to the labelled Eppendorf tubes as protein analysis was not necessary. Cell lysate samples were quantified for protein using the Pierce™ BCA assay using 30 μl aliquots so that the remaining total sample volumes would be consistent at 210 μl. Samples were then freeze‐dried and 50 μl of concentrated nitric acid 65% (v/v) was added for digestion overnight at room temperature (20–22°C). The next day, samples were heated at 90°C for 20 min to complete the digestion, leaving a volume after digestion of approximately 40 μl. To each sample, 960 μl of 1% (v/v) of nitric acid diluent was added to a final volume of 1 ml. The samples were injected onto an Agilent 7700 series ICP‐MS instrument (Agilent Technologies) under routine multi‐element operating conditions using a Helium Reaction Gas Cell. The instrument was calibrated using 0, 5, 10, 50, 100 and 500 ppb of certified multi‐element ICP‐MS standard calibration solutions (ICP‐MS‐CAL2‐1, ICP‐MS‐CAL‐3 and ICP‐MS‐CAL‐4, Accustandard, New Haven, CT) for a range of elements. A certified standard solution containing 200 ppb of Yttrium (Y89) was used as an internal control (ICP‐MS‐IS‐MIX1‐1, Accustandard). Raw ppb data of the elements were converted to μmol/L with dilution factors applied. Final concentrations of metals (μmol/L) were then blank corrected with treatment blanks and normalised to total protein obtained from the BCA assay. The final unit expressed as a percentage of the control was nmol of Cu/mg protein.
Corresponding organizations : Monash University, University of Melbourne, Florey Institute of Neuroscience and Mental Health
4
Quantifying Iron Content in Mouse Brain and Muscle
Iron content was assessed in samples via ICP-MS as reported previously [79 (link)]. Briefly, mouse brain cortices (n = 8/group) were digested overnight in concentrated nitric acid and then heated for 20 min at 90 °C. The volume of each sample was reduced to approximately 40 µL and then diluted to a final volume of 600 µL with 1% (v/v) nitric acid diluent. Measurements were performed using an Agilent 7700× series ICP-MS instrument.
Muscle samples were assessed for iron content using “microdroplet” laser ablation-ICP-MS (LA-ICP-MS) as described previously [80 (link)]. Briefly, muscle samples (n = 8/group) were homogenized in tris(hydroxymethyl)-aminomethane buffered saline (TBS)-based homogenization buffer as described previously [81 (link)]. Samples were assessed for protein content using the Pierce BCA Protein Assay kit (Thermo Fisher Scientific, Waltham, MA, USA) and then diluted to a consistent protein concentration. One microliter of each sample was pipetted onto a glass slide and left to air dry overnight. Droplet residues were ablated off the slide surface using laser ablation and analyzed using Iolite software [80 (link)]. Measurements were performed using an NWR-213 laser ablation unit (Electro Scientific Industries, Portland, OR, USA) hyphenated to an Agilent 8800 ICP-QQQ-MS.
Iron content was normalized within samples using a multielement control (Mg, P, and K for brain; C and P for muscle) and expressed relative to the WT-SED group.
Muscle samples were assessed for iron content using “microdroplet” laser ablation-ICP-MS (LA-ICP-MS) as described previously [80 (link)]. Briefly, muscle samples (n = 8/group) were homogenized in tris(hydroxymethyl)-aminomethane buffered saline (TBS)-based homogenization buffer as described previously [81 (link)]. Samples were assessed for protein content using the Pierce BCA Protein Assay kit (Thermo Fisher Scientific, Waltham, MA, USA) and then diluted to a consistent protein concentration. One microliter of each sample was pipetted onto a glass slide and left to air dry overnight. Droplet residues were ablated off the slide surface using laser ablation and analyzed using Iolite software [80 (link)]. Measurements were performed using an NWR-213 laser ablation unit (Electro Scientific Industries, Portland, OR, USA) hyphenated to an Agilent 8800 ICP-QQQ-MS.
Iron content was normalized within samples using a multielement control (Mg, P, and K for brain; C and P for muscle) and expressed relative to the WT-SED group.
Corresponding organizations : University of Eastern Finland, University of Melbourne, University of Technology Sydney, QIMR Berghofer Medical Research Institute
5
Quantitative Trace Element Analysis in Mouse Tissues
The tissue concentrations of trace elements were measured by an established and fully validated method using ICP-MS as described previously (Maynard et al. 2002 (link); Kanninen et al. 2013 (link)). Due to technical problems, four samples were not processed (Fmr1 KO cortex, WT liver and both Fmr1 KO and WT cerebellum). Briefly, weighed tissue samples were lyophilised, digested in 100–300 µl of 65% nitric acid (Merck, Kilsyth, Victoria, Australia) overnight at room temperature (RT), and heated for 20 min at 90 °C. Then an equivalent volume of 30% hydrogen peroxide (30% Aristar, BDH) was added and samples were incubated for 30 min at RT, followed by 15 min at 70 °C. The average reduced volume was determined, and the samples were diluted with 1% nitric acid diluent using dilution factors as shown in Table 1 . Volume of digest was the amount of the digested sample that was used to make the dilution.
Measurements were made with an Agilent 7700 series ICP-MS instrument (Agilent Technologies, Santa Clara, CA, USA) under routine multi-element operating conditions using helium as cell gas. The instrument was calibrated using 0, 5, 10, 50, and 100 ppb of certified multi-element standard calibration solutions (ICP-MS-CAL2-1, ICP-MS-CAL-3, and ICP-MS-CAL-4; Accustandard, New Haven, CT, USA) for the range of the elements and 200 ppb of Yttrium (Y89) was used as internal control (ICP-MS-IS-MIX1-1, Accustandard). Three media blanks were used to determine detection limits. Conversion of readings in ppb was performed as follows: (μg/g) = (raw ppb value × dilution factor × reduced digest volume)/(tissue wet weight g). Samples were analysed in triplicate and median values were used for analyses. Results are expressed in micrograms of metal per gram of wet (µg/g wet wt). The concentration of 23 minerals and trace elements were assessed in mouse heart, liver, spleen, and brain tissues. Altogether, 7 trace elements (Cu, Fe, K, Mg, Mn, Na, and P) were above detection limits in our set of samples (Table2 ), whereas 16 trace elements (Al, B, Ba, Ca, Cd, Co, Cr, Li, Mo, Ni, Rb, Ru, Se, Sr, Ti, and Zn) could not be reliably detected.
Measurements were made with an Agilent 7700 series ICP-MS instrument (Agilent Technologies, Santa Clara, CA, USA) under routine multi-element operating conditions using helium as cell gas. The instrument was calibrated using 0, 5, 10, 50, and 100 ppb of certified multi-element standard calibration solutions (ICP-MS-CAL2-1, ICP-MS-CAL-3, and ICP-MS-CAL-4; Accustandard, New Haven, CT, USA) for the range of the elements and 200 ppb of Yttrium (Y89) was used as internal control (ICP-MS-IS-MIX1-1, Accustandard). Three media blanks were used to determine detection limits. Conversion of readings in ppb was performed as follows: (μg/g) = (raw ppb value × dilution factor × reduced digest volume)/(tissue wet weight g). Samples were analysed in triplicate and median values were used for analyses. Results are expressed in micrograms of metal per gram of wet (µg/g wet wt). The concentration of 23 minerals and trace elements were assessed in mouse heart, liver, spleen, and brain tissues. Altogether, 7 trace elements (Cu, Fe, K, Mg, Mn, Na, and P) were above detection limits in our set of samples (Table
The absolute metal content in the set of WT tissue samples of average size and concentrations of the medium and acid blanks
| Metal | Cortex | Cerebellum | Liver | Spleen | Heart | Media blank | Acid |
|---|---|---|---|---|---|---|---|
| Na (µg) | 210 | 96 | 130 | 38 | 98 | 25 | 32 |
| Mg (µg) | 14 | 7.5 | 36 | 9.2 | 14 | 0.69 | 0.31 |
| P (µg) | 330 | 190 | 530 | 180 | 170 | 4.2 | 2.5 |
| K (µg) | 330 | 160 | 500 | 180 | 160 | 4.1 | 5.4 |
| Mn (ng) | 29 | 21 | 120 | 8.2 | 31 | 0.013 | 0 |
| Fe (ng) | 910 | 540 | 12,000 | 26,000 | 3500 | 0.29 | 0 |
| Cu (ng) | 300 | 200 | 720 | 36 | 360 | 0.076 | 0.0016 |
Corresponding organizations : University of Helsinki, University of Eastern Finland, University of Melbourne
Top 2 most cited protocols using «agilent 7700 series icp ms instrument»
1
Measuring pH-Dependent Iron Release
To measure pH-dependent release of iron from iron-binding proteins, 1-ml aliquots of 0.5 mg ml−1 holo-hemoglobin, ferritin, and transferrin were dissolved in ACCM-2 inorganic basal buffer (26 (link)) without FeSO4 at either pH 7.3 or 4.75. These solutions were incubated for 2 h at 37°C before being passed through a 3-kDa filter (Amicon Ultra-0.5 centrifugal filter unit; MilliporeSigma, Burlington, MA) to separate iron bound to intact proteins from the free iron. To process, 0.1 ml concentrated HNO3 was added to the filtrates to extract free iron overnight at room temperature, heated to 80°C for 1 h, and then diluted to a final volume of 5 ml with double-distilled water (ddH2O). Iron content was measured by inductively coupled plasma mass spectroscopy (ICP-MS) using an Agilent 7700 series ICP-MS instrument (Agilent Technologies, Santa Clara, CA).
Corresponding organizations : Washington State University
2
Quantifying Transition Metals in Mo-based POMs
ICP-MS analyses were carried out on an Agilent
7700 Series ICP-MS instrument. A calibration curve for the different
elements (Mo, Co, Mn, Fe) was established by measuring different concentrations
of standard solutions of these elements. For sample preparation, samples
of Mx⊂Q{Mo132PO4} were prepared by weighing the sample into a quartz test
tube and heated at 600 °C in air for 2 h to volatilize and remove
all of the organic component. After the sample was cooled, 2 M NaOH
(0.5 mL) was added, followed by DDW and then 0.5 mL of high purity
concentrated HNO3. The sample was diluted to the desired
concentration and filtered with 0.2 μm filter. There was some
variation (±5%) from the preparation of the Mx⊂Q{Mo132PO4} samples, but overall
the encapsulated amount of M2+ was about 75% of the nominal
amount of transition metal salt that was added.
7700 Series ICP-MS instrument. A calibration curve for the different
elements (Mo, Co, Mn, Fe) was established by measuring different concentrations
of standard solutions of these elements. For sample preparation, samples
of Mx⊂Q{Mo132PO4} were prepared by weighing the sample into a quartz test
tube and heated at 600 °C in air for 2 h to volatilize and remove
all of the organic component. After the sample was cooled, 2 M NaOH
(0.5 mL) was added, followed by DDW and then 0.5 mL of high purity
concentrated HNO3. The sample was diluted to the desired
concentration and filtered with 0.2 μm filter. There was some
variation (±5%) from the preparation of the Mx⊂Q{Mo132PO4} samples, but overall
the encapsulated amount of M2+ was about 75% of the nominal
amount of transition metal salt that was added.
Corresponding organizations : Weizmann Institute of Science
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