A sterile filtered sample (200 μl) of nutlin-3a ND was applied to a Zorbax GF-250 column equilibrated in PBS containing additionally 0.15 M NaCl. Chromatography was performed on a Perkin-Elmer Series 200 System at a flow rate of 1 ml/min. Fractions (0.5 ml) were collected over a period of 30 min. For comparison, a sample containing empty ND was applied to the column.
Series 200 system
Manufactured by PerkinElmer
Sourced in Italy, United States
The Series 200 System is a versatile and reliable lab equipment designed for a wide range of applications. It features a modular architecture that allows for customization to meet specific needs. The core function of the Series 200 System is to provide precise and accurate analytical measurements in a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Aanalyst 100 system, by PerkinElmer (1 mentions)
Silica gel 60 f254, by Merck Group (1 mentions)
Elan drc 2, by PerkinElmer (1 mentions)
320 s ph meter, by Mettler Toledo (1 mentions)
Zorbax eclipse xdb c8 column, by Agilent Technologies (1 mentions)
Scopoletin, by Merck Group (1 mentions)
Medicarpin, by Merck Group (1 mentions)
Maackiain, by Merck Group (1 mentions)
Flexar, by PerkinElmer (1 mentions)
Totalchrom workstation software, by PerkinElmer (1 mentions)
12 protocols using series 200 system
1
Purification of Nutlin-3a Nanoparticles
2
Synthesis and Characterization of Porphyrin Derivatives
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All reagents and deuterated solvents used for synthesis were of reagent grade or better and were used without further purification unless stated otherwise. Starting materials, reagents and deuterated solvents were purchased from Sigma Aldrich, and all other solvents were purchased from Caledon Laboratories. The PNH2 precursor, 5-(4-aminophenyl)-10, 15, 20-(tri-4-sulfonatophenyl)porphyrin triammonium, was purchased from PorphyChem. All reactions were carried out under argon. Thin layer chromatography was carried out on pre-coated aluminum plates of Silica Gel 60 F254 from Merck. Column chromatography was performed using Caledon Silica Gel 60. Dialysis was performed with Biotech CE dialysis tubing (MWCO 100–500 Da). Cation exchange was performed using an Aberlite IR120 H resin. All spectroscopic data for structural characterizations were obtained using the research facilities in the Department of Chemistry. NMR spectra were recorded on a Brucker-500 MHz. UV-visible spectra were recorded on an Agilent 8453 system. HPLC spectra were recorded on a PerkinElmer SERIES 200 system. FAA spectra were recorded on a PerkinElmer AAnalyst 100 system. Mass spectroscopy was carried out on a Agilent 6538 Q-TOF system.
3
Purification of Nutlin-3a Nanoparticles
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A sterile filtered sample (200 μl) of nutlin-3a ND was applied to a Zorbax GF-250 column equilibrated in PBS containing additionally 0.15 M NaCl. Chromatography was performed on a Perkin-Elmer Series 200 System at a flow rate of 1 ml/min. Fractions (0.5 ml) were collected over a period of 30 min. For comparison, a sample containing empty ND was applied to the column.
4
Seaweed Arsenic Analysis using ICP-MS
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The seaweed samples were digested using a microwave-assisted digestion system (CEM Mars 40, USA) and the total arsenic levels were determined using an inductively coupled plasma mass spectrometer (ICP-MS) (PerkinElmer, ELAN DRC II, USA). The arsenic species were analyzed by HPLC using a Series 200 System (PerkinElmer) coupled to an ICP-MS system. The pH values were measured using a Mettler Toledo 320-S pH-meter (Mettler Toledo Co., China). A temperature-consistent oscillating water bath (SHA-B, GuoHua Co., China) and a centrifuge (Eppendorf 5810R, Germany) were used.
5
HPLC Determination of Norfloxacin Release
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Norfloxacin released from each scaffold was determined by DAD–HPLC (Series 200 system, PerkinElmer, Milan, Italy). A Zorbax Eclipse XDB-C8 column (4.6 mm × 150 mm, silica particle size 5 μm, Agilent, Milan, Italy) was used as the stationary phase. The mobile phase was based on acetonitrile/methanol/citric acid 0.4 M, 7:15:78 (% v/v) at a flow rate of 1.0 mL/min, using 275 nm wavelength detection [21 (link),22 (link)]. The injection volume was 10 μL. Calibration curves were obtained using norfloxacin standard solution in the mobile phase, in saline solution or processed as the samples subjected to lysozyme degradation. In every case, the method was linear from 0.08 to 200 μg/mL with an R2 value that was always higher than 0.995.
6
HPLC-DAD Analysis of Olive Oil Compounds
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HPLC-DAD analysis was performed according to the previously described procedure [69 (link),70 (link)]. A Perkin Elmer Series 200 system (PerkinElmer, Waltham, MA, USA) with diode array detector was used, equipped with a C18 Restek column (5 μm, 250 × 4.0 mm). The gradient change of a mobile phase consisting of two components was used (A: 2% acetic acid and B: methanol) with an increasing percentage of B as follows: 5% B for 2 min; going to 25% B till 10 min; 40% B till 20 min; 50% B till 30 min; and 100% B till the end of the run at 45 min. Between two runs, the column was washed for 15 min with methanol. The flow rate was 1 mL/min and the injection volume was 25 μL at 25 °C. The absorbance was detected at 278 nm. The retention times of the analysed compounds were compared with those of standards (min): hydroxytyrosol (12.20), tyrosol (15.66), cinnamic acid (30.20), pinoresinol (33.41), and apigenin (42.65). The concentrations were calculated using calibration curves (Table S3 ).
7
HPLC Analysis of Radiolabeled Compounds
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Both the crude reaction mixtures and purified radiolabeled preparations were analyzed by RP-HPLC on a Perkin Elmer Series 200 system. Analysis conditions were as follows: flow rate: 1 mL/min; mobile phase: 0.1% TFA in water (buffer A) and 0.1% of TFA in acetonitrile (buffer B); gradient: 1 min with 100 % A, then to 90:10 A/B over 2 min, followed by 70:30 A/B over 20 min. Radioactivity of the eluate was monitored using a NaI “flow” radiodetector (Bioscan, Santa Barbara, CA, United States) connected in-line, whereas cold products were detected using an integrated photodiode array detector by setting the absorbance at 220 nm.
8
Purification and Immunoblot of Wnt3a
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Wnt3a ND (250 µl) were concentrated by centrifugal filtration (Centricon 50 kDa MWCO) to 70 µl and subjected to HPLC on a 9.4 × 250 mm Zorbax GF-250 column equilibrated in PBS plus 0.15 M NaCl. Chromatography was performed on a Perkin-Elmer Series 200 System at a flow rate of 1 ml/min. A portion (150 µl) of selected fractions was precipitated with chloroform/methanol after addition of 10 µg bovine serum albumin [20 (link)]. Pelleted material was solubilized in electrophoresis buffer and subjected to anti-Wnt3a immunoblot.
9
Phytoalexin Quantification by HPLC
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Phytoalexins were identified and quantified by HPLC using an HPLC (Perkin Elmer, Series 200 System, Monza, Italy) equipped with a diode array detector set to 210 and 220 nm.
Analyses were carried out in duplicate injecting 20 μL of all samples for each analysis.
Standards (scopoletin, pisatin, medicarpin, maackiain (Sigma-Aldrich, St. Luis, MO, USA)) were prepared at concentration of 1 mg ml -1 in methanol and diluted in a 1:10 ratio in a C 18 column 250×4 mm internal diam., containing 5 mm diam. beads (LiChrospher 100 RP-18), using a flow rate of 1 ml min -1 and elution with CH 3 CN/H 2 O along a linear gradient of CH 3 CN from 20 to 33% over a period of 90 min, and again from 33 to 20% for 5 min. The column was re-equilibrated under isocratic conditions for 5 min before the next run. The data obtained were expressed in µg g -1 FW.
Analyses were carried out in duplicate injecting 20 μL of all samples for each analysis.
Standards (scopoletin, pisatin, medicarpin, maackiain (Sigma-Aldrich, St. Luis, MO, USA)) were prepared at concentration of 1 mg ml -1 in methanol and diluted in a 1:10 ratio in a C 18 column 250×4 mm internal diam., containing 5 mm diam. beads (LiChrospher 100 RP-18), using a flow rate of 1 ml min -1 and elution with CH 3 CN/H 2 O along a linear gradient of CH 3 CN from 20 to 33% over a period of 90 min, and again from 33 to 20% for 5 min. The column was re-equilibrated under isocratic conditions for 5 min before the next run. The data obtained were expressed in µg g -1 FW.
10
Simultaneous HPLC-UV Quantification of Scopolamine and Atropine
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Scopolamine and atropine obtained from a raw extract of D. stramonium L. were simultaneously identified by HPLC-UV method (Perkin-Elmer Series 200 system with a UV/Vis detector). For the quantitative determination of the two compounds, HPLC was coupled with full-scan diode array spectrophotometry (DAD). HPLC method used a 100-5C8 Kromasil column (250 × 4.6 mm) with gradient elution and a working temperature of 25 °C in the column. The mobile phase was composed of acetonitrile (25%) and an aqueous solution (75%) (5 mM sodium 1-heptanesulfonate monohydrate, pH = 3.5). Detection was performed at UV (λ = 230 ± 4 nm with reference λ = 360 ± 8 nm). The calibration curve was linear between 0.13-13.75 mg/mL (r = 0.9951, n = 8) for scopolamine and 0.25-25.5 mg/mL (r = 0.9999, n = 8) for atropine (Hinescu 2011) . The data were generated by ChromeGate, using atropine and (-) scopolamine as standard samples.
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