The content of inorganic ion was determined by ion exchange chromatography (Dionex DX120; Dionex Corporation, Sunnyvale, CA, USA) with a conductivity detector, as reported by D’Imperio et al. [17 (link)]. The content of Na+, K+, Mg2+, and Ca2+ was determined in 1 g of dried sample, using an IonPac CG12A guard column and an IonPac CS12A analytical column (Dionex Corporation); the elution was performed with 18 mM of methanesulfonic acid (Thermo Scientific™ Dionex™, Waltham, MA, USA). Peaks identification and calibration were performed using the Multi Element IC Standard solution Fluka TraceCERT®, Supelco® (Merck KGaA, Darmstadt, Germany). The contents of Cl− and NO3− were determined in 0.5 g of dried sample using an IonPac AG14 precolumn and an IonPac AS14 separation column (Dionex Corporation). The eluent consisted of 3.5 mmol·L−1 of sodium-carbonate (Thermo Scientific™ Dionex™, USA) and 1.0 mmol·L−1 of sodium-bicarbonate solution (Thermo Scientific™ Dionex™, USA), and 50 mL of the same eluent was used to extract the anions. Inorganic cation content determination was carried out in triplicate. Peaks identification and calibration were performed using the Multi Element IC Standard sol. IC-MAN-18 (6E) of Chem-Lab (Palin Corporation, Elderslie, UK).
Dionex
Manufactured by Thermo Fisher Scientific
Sourced in United States, France, Canada, Italy, Belgium
Dionex is a suite of analytical instruments and software developed by Thermo Fisher Scientific. The core function of Dionex products is to enable chromatographic separation, detection, and data analysis for a variety of applications in scientific research and industrial processes. Dionex systems are designed to provide reliable and accurate analytical results.
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Lab products found in correlation
Superdex 200 increase 10 300 gl column, by GE Healthcare (4 mentions)
T rex, by Wyatt Technology (3 mentions)
Ultimate 3000, by Thermo Fisher Scientific (3 mentions)
Xylose, by Merck Group (2 mentions)
Xylo oligosaccharides, by Megazyme (2 mentions)
Carbopac pa 100 column, by Thermo Fisher Scientific (2 mentions)
Dad 3000, by Thermo Fisher Scientific (2 mentions)
Dionex ultimate 3000, by Thermo Fisher Scientific (2 mentions)
Dawn8 heleos 8, by Wyatt Technology (2 mentions)
Delsamax pro, by Beckman Coulter (2 mentions)
Dawn8 heleos, by Wyatt Technology (1 mentions)
Beechwood xylan, by Megazyme (1 mentions)
Ysi 2700 glucose analyzer, by Yellow Springs Instruments (1 mentions)
Lc ms grade, by Thermo Fisher Scientific (1 mentions)
Ultimate 3000 rslc, by Thermo Fisher Scientific (1 mentions)
Wps 3000tsl, by Thermo Fisher Scientific (1 mentions)
Aa 7000, by Shimadzu (1 mentions)
Elisa, by Calbiotech (1 mentions)
Elisa, by DRG International (1 mentions)
Licrospher 100 rp 18, by Merck Group (1 mentions)
58 protocols using dionex
1
Inorganic Ion Analysis by Ion Chromatography
2
Urine Electrolytes and Bone Biomarkers Analysis
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Urine electrolytes were measured by ion chromatography (Dionex Aquion Ion Chromatography System, Thermo Fisher Scientific). Samples were diluted 1:50 in ddH2O with eluent of 4.5 m m Na2CO3/1.5 m m NaHCO3 in ddH2O for anions and 20 m m Methanesulfonic acid in ddH2O for cations. Results were interpolated from a standard curved generated by serial dilutions of Dionex 5 anion and 6 cation-I standards (Dionex, Thermo Fisher Scientific Inc., Mississauga, ON, Canada) using Chromeleon 7 Chromatography Data System software (Thermo Scientific). Urine creatinine was measured using the Parameter creatinine kit (R&D systems, Minneapolis, MN). Plasma PTH (Immutopics Mouse Intact PTH 1–84), 1,25(OH)2-vitamin D (Immunodiagnostic Systems Limited, Boldon, UK), FGF23 (Kainos Laboratories, Inc.) and milk EGF (Quantikine Human EGF Immunoassay, R&D systems) were measured by ELISA.
3
Automated Production and Quality Control of [18F]FDG
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The 18F-FDG was synthesized using a cassette-based GE Fastlab synthesis module (GE Healthcare, Belgium). 18F-fluoride was produced by bombarding 18O-enriched water using a 11-MeV proton beam in an Eclips HP cyclotron (Siemens, Knoxville, Tennessee). The purified 18F-FDG was then diluted with 0.9% NaCl (Baxter, Belgium) and sterile filtered through a 0.22 µmol/L filter. Quality control was performed according to European Pharmacopoeia 7.1. Radiochemical identity was confirmed by high-performance liquid chromatography (HPLC; Dionex; ThermoFisher, Belgium) and radio thin-layer chromatography. Radiochemical purity was determined by HPLC (Dionex, ThermoFisher, Belgium) and ranged from a minimum of 99.56% to a maximum of 99.97%. Radionuclidic identity and purity were confirmed by gamma spectrum analysis (Multi-channel analyzer, Canberra).
4
SEC-MALS Analysis of Sis0455 Protein
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SEC-MALS experiments were performed using a Dionex (Thermo Scientific) HPLC system connected in-line to a UV detector (Thermo Scientific Dionex Ultimate 3000, MWD-3000), a Wyatt Dawn8 + Heleos 8-angle light-scattering detector and a Wyatt Optilab T-rEX refractive index detector. SEC was performed using a Superdex 200 Increase 10/300 GL column (GE Healthcare) at 20°C in a buffer containing 50 mM HEPES pH 8.0, 300 mM KCl. For the analysis, 50 μl of Sis0455 were injected at 5.0 mg/ml and 0.5 ml/min flow rate. ASTRA (version 8.0.2.5) software was used to collect the data from the UV, refractive index, and light scattering detectors. The weight average molecular masses, Mw, were determined across the elution profile from static LS measurements using ASTRA software and a Zimm model, which relates the amount of scattered light to the weight average molecular weight of the solute, the concentration of the sample, and the square of the refractive index increment (dn/dc) of the sample.
5
Production and Analysis of XOS
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The production of XOS was carried out using as a substrate commercial beechwood xylan (Megazyme) at a 1% concentration in a 50 mM Tris–HCl buffer, pH 9.0. Xylanases, Xyn11_Ec or Xyn11_Nb, were added at 0.75 µg/mL and incubated at 90ºC for different times. The reactions were stopped by placing the tubes on ice. XOS were analyzed by exchange ion chromatography, using a Dionex instrument equipped with a CarboPac PA100 column and a pulsed amperometric detector (Dionex, Thermo Fisher Scientific). Xylose (Sigma-Aldrich) and xylooligosaccharides, from two to six units (Megazyme), were used as the chromatographic standards.
6
Quantifying Leaf Metabolites in Plants
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For determining the content of titratable acids, aliquots of methanol extracts were titrated against NaOH (0.05 mm ) to a neutral endpoint and expressed as millimoles of H+ per gram fresh weight (fwt), as described by Cushman et al. (2008a (link)). Starch was extracted from the leaf mesophyll as described previously (Haider et al., 2012 (link)), and its content in mesophyll was measured as glucose equivalents using the colorimetric phenol/sulphuric acid test described by DuBois et al. (1956) . To determine starch content in guard cells, the peels were fixed (50 % v/v methanol, 10 % v/v acetic acid) and stained with Lugol’s iodine solution as described by Horrer et al. (2016) (link). Soluble sugars were measured in both tissues using the high-pressure ion chromatography (HPIC) technique (Thermo Scientific Dionex), and the amount of sugars (in micromoles per gram fresh weight) was calculated based upon standards of glucose, fructose, sucrose and maltose. Malate content in both mesophyll and guard cell-enriched epidermis was determined by the enzymatic method developed by Hohorst (1970) .
7
Fermentation Glucose Quantification by HPLC
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During the fermentation, culture supernatant was obtained by centrifuging (MiniSpin Eppendorf, Hamburg, Germany) fermentation broth samples in Eppendorf tubes at 12,000 g for 3 min. The glucose concentration in supernatant was measured with a YSI 2700 glucose analyzer (Yellow Springs Instruments, Yellow Springs, OH, USA). Glucose and organic acid concentrations of filtered supernatants (Minisart filters 0.2 µm size pore, Sartorius, Göttingen, Germany) were determined by HPLC (Ultimate 3000, Dionex, Thermo Fisher Scientific, Courtaboeuf, France) using an Aminex HPX-87H+ column (Bio-Rad Life Science, Marnes-la-Coquette, France) under the following conditions: 50 °C, 5 mM H2SO4 as mobile phase at a flow rate of 0.5 mL·min−1 and detection with a refractometer and an UV detector at 210 nm. The samples were previously diluted in deionised water. External standards (0.2 to 5 g·L−1) were used for compound identification and quantification. Chromatograms were analyzed with Chromoleon software 6.80 (Thermo Fisher Scientific, Courtaboeuf, France). For the flask experiments, the glucose uptake and the biomass yield were calculated at the end of the growth phase.
8
Nano-HPLC Peptide Separation Protocol
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Following digestion, peptides were separated by a nano high performance liquid chromatography system (Ultimate 3000 RSLC) using a pre-concentration trap column (Acclaim® PepMap™ μ-Precolumn) and a nano separation column (Acclaim® PepMap™ RSLC 75 μm × 25 cm, nano Viper C18, 2 μm, 100 Å) (both Dionex™, Thermo Fisher Scientific, USA). The mobile phases used were water (LC/MS grade Fisher Scientific, USA) with 0.1% formic acid (A) and 80% acetonitrile (ACN) with 0.1% formic acid (B). A mobile phase gradient from 4% B to 35% B in 120 min and then up to 90% B in 15 min followed by a washing step with 90% B for 10 min was run.
9
Ultrafiltration Centrifugation for FH Entrapment Efficiency
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Drug EE% was determined
by the ultrafiltration centrifugation method.19 (link) Briefly, 1 mL of FH-HEX was centrifuged in an Amicon centrifugal
tube (100 000 molecular weight cutoff (MWCO), Millipore) at
15 000 rpm for 15 min at 4 °C. The supernatant was then
collected, and the amount of free FH was quantified using a previously
validated HPLC (Dionex, Thermo Scientific), equipped with an LPG-3400SD
quaternary pump, a WPS-3000TSL autosampler, a variable-wavelength
detector (VWD-3000), and a reverse-phase C18 column (Hypersil
ODS, 150 × 4.6 mm2, 5 μm). The wavelength of
the UV detector was set at 226 nm. The samples were separated using
acetonitrile and deionized water containing 10 mM aqueous triethylamine
at a ratio of 55:45 v/v at a flow rate of 1 mL/min at 25 °C.
The coefficient of determination (R2)
of the FH calibration curve in PBS (pH 7.4) in the concentration range
of 1–100 μg/mL was 0.9994, and the respective limits
of detection (LOD) and quantification (LOQ) were 0.5 and 1 μg/mL.
The coefficients of variation (CV)% ranged from 2.1 to 4.9%, and the
accuracy for FH determination was 1.5–4.6% with a mean% drug
recovery of 97.5 ± 1.16%.
The adsorbed amount of FH on
the Amicon centrifugal tube membrane was determined by filtering an
FH solution of the same concentration, and then the amount of free
FH in the filtrate was quantified.56 (link)EE% of FH was calculated using the following equation:
by the ultrafiltration centrifugation method.19 (link) Briefly, 1 mL of FH-HEX was centrifuged in an Amicon centrifugal
tube (100 000 molecular weight cutoff (MWCO), Millipore) at
15 000 rpm for 15 min at 4 °C. The supernatant was then
collected, and the amount of free FH was quantified using a previously
validated HPLC (Dionex, Thermo Scientific), equipped with an LPG-3400SD
quaternary pump, a WPS-3000TSL autosampler, a variable-wavelength
detector (VWD-3000), and a reverse-phase C18 column (Hypersil
ODS, 150 × 4.6 mm2, 5 μm). The wavelength of
the UV detector was set at 226 nm. The samples were separated using
acetonitrile and deionized water containing 10 mM aqueous triethylamine
at a ratio of 55:45 v/v at a flow rate of 1 mL/min at 25 °C.
The coefficient of determination (R2)
of the FH calibration curve in PBS (pH 7.4) in the concentration range
of 1–100 μg/mL was 0.9994, and the respective limits
of detection (LOD) and quantification (LOQ) were 0.5 and 1 μg/mL.
The coefficients of variation (CV)% ranged from 2.1 to 4.9%, and the
accuracy for FH determination was 1.5–4.6% with a mean% drug
recovery of 97.5 ± 1.16%.
The adsorbed amount of FH on
the Amicon centrifugal tube membrane was determined by filtering an
FH solution of the same concentration, and then the amount of free
FH in the filtrate was quantified.56 (link)EE% of FH was calculated using the following equation:
10
Quantification of Ammonium Sulfate Aerosols
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