Mass spectrometric characterizations of fresh vs. stored 10 mM stock solutions of CM572 were carried out in the Brown University Mass Spectrometry Facility on the Agilent 6530 Accurate Mass Q-TOF LC-MS system (Agilent Technologies, Santa Clara, CA). The Agilent 1260 Infinity series LC is comprised of a HiP-ALS autosampler, Bin Pump, and TCC column oven. HPLC-MS analysis was carried out with an Agilent column Zorbax C18, 2.1 × 50 mm, with mobile phases A = 5% acetonitrile/95% H2O/0.1% formic acid and B = acetonitrile/0.1% formic acid and the following time program of the gradient: 0 min 5% B, linear to 65% B at 8 min, to 95% B at 9 min, back to 5% B at 11 min and equilibrate for 10 min. The Q-TOF MS instrument was operated with an electrospray ion source in the positive ionization mode, mass range 100 – 3200 m/z, and 1 spectra/sec acquisition rate. Data analysis was performed using MassHunter Workstation Software Qualitative Analysis Version B.06.00 (Agilent Technologies, Santa Clara, CA).
1260 infinity series lc
Manufactured by Agilent Technologies
The Agilent 1260 Infinity series LC is a high-performance liquid chromatography (HPLC) system designed for analytical laboratory applications. It provides reliable and consistent performance for a variety of separation tasks. The 1260 Infinity series LC features modular components, including pumps, autosampler, column compartment, and detectors, allowing for customization to meet specific analytical requirements.
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Lab products found in correlation
Agilent 6530 accurate mass q tof lc ms system, by Agilent Technologies (1 mentions)
Masshunter workstation software qualitative analysis version b 06, by Agilent Technologies (1 mentions)
7700 series icp ms, by Agilent Technologies (1 mentions)
Glass microfibre filter, by Cytiva (1 mentions)
Zorbax rx c8, by Agilent Technologies (1 mentions)
Whatman, by Cytiva (1 mentions)
3 protocols using 1260 infinity series lc
1
Characterization of CM572 Compound
2
Quantification of HSPC and Cholesterol in Liposomes
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Quantification of HSPC and cholesterol was performed using an Agilent 1260 infinity series LC equipped with an Agilent evaporative light scattering detector (ELSD) operating at a 1.6 L/min gas flow with the evaporator and nebulizer heated to 80 °C and 50 °C, respectively. Lipids were extracted from liposome samples by direct dilution with methanol. Chromatographic separation of HSPC and cholesterol was achieved using an Eclipse XDB C18 column (150 × 4.6 mm, 5 μM) at 40 °C with mobile phase composed of methanol (A) and water (B) both with 0.1% trifluoroacetic acid (v/v). The initial mobile phase was 90% A with a flow rate of 1.0 mL/min, which was gradually increased to 100% A over 4 min. Following an 11-min equilibration, the composition was rapidly changed back to 90% A where it was maintained for the remaining 5 min of the run.
DL ratios were calculated using the following equation:
DL ratios were calculated using the following equation:
3
Selenium Speciation Analysis by HPLC-ICP-MS
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Separations were performed on an Agilent Zorbax RX-C8 4.6 × 250 mm 5 μm column using an Agilent Technologies 1260 infinity series LC system connected to an Agilent Technologies 7700× series ICP-MS. The mobile phase was water/methanol/TFA (97.9:2.0:0.1) at a flow rate of 1 mL/min, and the column temperature was set at 30 °C.
A Heinemann 130W ultrasonic-homogeniser (HTU SONI 130, USA) equipped with a 3 mm double step titanium probe was used for sample preparation. A CEM Discover microwave equipped with an Explorer SP-D Plus 24/48 autosampler was used for sample extraction. An ELGA Purelab Flex S7 HPLC water system was used to produce >18 MΩ cm water, and samples were filtered using Chromafil Xtra RC-20/25. The HPLC mobile phase was filtered with Whatman Glass Microfibre Filters.
SeMet standard powder was obtained from Sigma (>98 % by TLC). This was used to prepare a 100 mg/L stock solution in 0.1 M HCl. Aliquots (0.1 mL) of this solution were frozen and used fresh each day.
The instrument was calibrated over the range of 50 to 250 ppb SeMet (20.15 to 100.75 ppb Se as SeMet) as described previously [19] .
A Heinemann 130W ultrasonic-homogeniser (HTU SONI 130, USA) equipped with a 3 mm double step titanium probe was used for sample preparation. A CEM Discover microwave equipped with an Explorer SP-D Plus 24/48 autosampler was used for sample extraction. An ELGA Purelab Flex S7 HPLC water system was used to produce >18 MΩ cm water, and samples were filtered using Chromafil Xtra RC-20/25. The HPLC mobile phase was filtered with Whatman Glass Microfibre Filters.
SeMet standard powder was obtained from Sigma (>98 % by TLC). This was used to prepare a 100 mg/L stock solution in 0.1 M HCl. Aliquots (0.1 mL) of this solution were frozen and used fresh each day.
The instrument was calibrated over the range of 50 to 250 ppb SeMet (20.15 to 100.75 ppb Se as SeMet) as described previously [19] .
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