SEC-MALS experiments were performed using a Superdex Increase 200 10/300 GL column (GE Healthcare) on an Agilent 1260 HPLC Infinity II in PBS buffer at RT (~297 K). Protein elution was monitored by three detectors in series namely, an Agilent multi-wavelength absorbance detector (absorbance at 280 nm and 254 nm), a Wyatt miniDAWN TREOS multiangle light scattering (MALS) detector, and a Wyatt Optilab rEX differential refractive index (dRI) detector. The column was pre-equilibrated overnight in the running buffer to obtain stable baseline signals from the detectors before data collection. Molar mass, elution concentration, and mass distributions of the samples were calculated using the ASTRA 7.1.3 software (Wyatt Technology). A BSA solution (2–4 mg/ml), purchased from Sigma-Aldrich and directly used without further purification, was used to calibrate inter-detector delay volumes, band broadening corrections, and light-scattering detector normalization using standard protocols within ASTRA 7.1.3.
1260 infinity 2 hplc
Manufactured by Agilent Technologies
Sourced in United States
The 1260 Infinity II HPLC is a high-performance liquid chromatography system manufactured by Agilent Technologies. It is designed to provide reliable and accurate separation and analysis of a wide range of chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Astra 7, by Wyatt Technology (3 mentions)
Minidawn treos 2 mals detector, by Wyatt Technology (3 mentions)
Optilab t rex refractive index detector, by Wyatt Technology (3 mentions)
Bsa solution, by Merck Group (2 mentions)
Multi wavelength absorbance detector, by Wyatt Technology (2 mentions)
Optilab rex differential refractive index, by Wyatt Technology (2 mentions)
Minidawn treos multi angle light scattering, by Wyatt Technology (2 mentions)
Superdex 200 increase 10 300 gl column, by GE Healthcare (2 mentions)
Superdex 200 increase column, by Cytiva (2 mentions)
Dulbecco s phosphate buffered saline dpbs, by Thermo Fisher Scientific (2 mentions)
Pm100usb power and energy meter, by Thorlabs (2 mentions)
Cm100 transmission electron microscope, by Philips (2 mentions)
Spectramax m4, by Molecular Devices (2 mentions)
Ultrospec 10 cell density meter, by GE Healthcare (2 mentions)
Eclipse c18 column, by Agilent Technologies (1 mentions)
Openlab cds chemstation, by Agilent Technologies (1 mentions)
Ellagic acid, by Merck Group (1 mentions)
Agilent 6530 q tof lc ms system, by Agilent Technologies (1 mentions)
8860 gc 5977bmsd, by Agilent Technologies (1 mentions)
Analytical balance, by Sartorius (1 mentions)
49 protocols using 1260 infinity 2 hplc
1
SEC-MALS Analysis of Biomolecular Samples
2
SEC-MALS Protein Characterization Protocol
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SEC-MALS experiments
were performed using a Superdex Increase 200
10/300 GL column (GE Healthcare) on an Agilent 1260 HPLC Infinity
II in phosphate buffer (PBS or 50 mM KPi KCl pH 7.4) at RT (ca. 297
K). Protein elution was monitored by three detectors in series namely,
an Agilent multiwavelength absorbance detector (absorbance at 280
and 254 nm), a Wyatt miniDAWN TREOS multiangle light scattering (MALS)
detector, and a Wyatt Optilab rEX differential refractive index (dRI)
detector. The column was pre-equilibrated overnight in running buffer
to obtain stable baseline signals from the detectors before data collection.
Molar mass, elution concentration, and mass distributions of the samples
were calculated using the ASTRA 7.1.3 software (Wyatt Technology).
A BSA solution (2–4 mg/mL), purchased from Sigma-Aldrich and
directly used without further purification, was used to calibrate
interdetector delay volumes, band broadening corrections, and light-scattering
detector normalization using standard protocols within ASTRA 7.1.3.
were performed using a Superdex Increase 200
10/300 GL column (GE Healthcare) on an Agilent 1260 HPLC Infinity
II in phosphate buffer (PBS or 50 mM KPi KCl pH 7.4) at RT (ca. 297
K). Protein elution was monitored by three detectors in series namely,
an Agilent multiwavelength absorbance detector (absorbance at 280
and 254 nm), a Wyatt miniDAWN TREOS multiangle light scattering (MALS)
detector, and a Wyatt Optilab rEX differential refractive index (dRI)
detector. The column was pre-equilibrated overnight in running buffer
to obtain stable baseline signals from the detectors before data collection.
Molar mass, elution concentration, and mass distributions of the samples
were calculated using the ASTRA 7.1.3 software (Wyatt Technology).
A BSA solution (2–4 mg/mL), purchased from Sigma-Aldrich and
directly used without further purification, was used to calibrate
interdetector delay volumes, band broadening corrections, and light-scattering
detector normalization using standard protocols within ASTRA 7.1.3.
3
HPLC Analysis of Drug Impurities
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The Agilent HPLC 1260 Infinity-II consists four channels, pressure range up to 600 bar, degasser with integrated purge valve, thermostatic sampler, and column compartment. The photo-diode array detector (PDA) connected to empower 3 software (Build 3471 SPs Installed: Feature Release 3 DB ID: 2639633283) to monitor the output signal. The column is X-terra phenyl, 250 × 4.6 mm, 5 µm (Part No: 186001147). Sartorius semi-micro and micro-balances were used for weighing of impurities, standards, and samples. Bio-technics ultra sonicator was used to extract drug from the sample matrix.
4
HPLC Analysis of Ellagic Acid
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The HPLC analysis was performed as previously described with minor modifications [16 (link),17 ]. HPLC analysis used an HPLC 1260 Infinity II (Agilent Technologies, Inc., Santa Clara, CA, USA) system with an Eclipse C18 column (2.1 × 150 mm, 3.5 μm; Agilent Technologies) under 40 °C. Ellagic acid (>95% purity, Sigma-Aldrich, CA, USA) was used as a marker. The mobile phases were 0.1% formic acid (A) in DW and acetonitrile (B). A flow rate of 0.12 mL/min was used, with the gradient condition as flow with a of flow rate: 95–70% A, 0–30 min; 70–5% A, 30–56 min; 30–0% B, 56–57 min; 0–95% B, 57–58 min; and 95% A, 58–70 min. The injection volume was 2 μL, and the UV wavelength was set at 275 nm. Data acquisition was performed by using the Agilent OpenLab CDS ChemStation (version 2.15.26; Santa Clara, CA, USA).
5
LC-MS Analysis of Chemical Compounds
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LC-MS analysis was conducted on an Agilent HPLC 1260 Infinity II coupled with an Agilent 6530 Q-TOF-LC-MS system (Agilent Technologies, Palo Alto, CA, USA). The HPLC column was a reversed-phase ZORBAX RRHT Extend-C18, 2.1 by 50 mm, 1.8 μm (Agilent Technologies, USA). The mobile phase was a gradient of 0.1% formic acid in water (A) and acetonitrile (B) at a flow rate of 0.2 mL/min. A linear gradient was initiated with 2% acetonitrile and linearly increased to 95% at 5.1 to 10 min. The flow rate was 0.2 mL/min, and the injection volume was 5 μL. Mass spectra in the m/z range 111 to 3,000 were obtained by positive ion (positive electrospray ionization) modes. The mass spectrometry conditions were the following: gas temperature, 300°C; N2 flow rate, 7 liters/min; nebulizer gas pressure, 35 lb/in2; capillary voltage, 3,500 V; fragmentor potentials, 175 V; Vcap, 3,500 V; skimmer, 65 V; and octopole RFPeak, 750 V. Data acquisition and analysis were conducted using Agilent LC-MS-QTOF MassHunter Data Acquisition Software version 10.1 and Agilent MassHunter Qualitative Analysis Software version 10.0, respectively (Agilent Technologies, USA).
6
Quantification of Paeoniflorin in Herbal Extracts
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Determination of TGP content in PLR-U and PLR-F was performed by alkaline hydrolysis - HPLC [11 ]. HPLC analysis was conducted using a HPLC 1260 Infinity II (Agilent Technologies, USA), and the C18 chromatographic column (250 mm × 4.6 mm, 5 μm, Elite Analytical Instruments Co., Dalian, China) was maintained at 35 °C, with 0.05 mol L−1 potassium dihydrogen phosphate-acetic acid (350:8, mobile phase A) and methanol-isopropanol (100:6, mobile phase B) as the mobile phase with a flow rate of 0.8 mL min−1, A:B = 70:30, and the detection wavelength utilized was set at 230 nm. Appropriate amount of fermentation broth was evaporated to dryness, and hydrolyzed by adding 10 mL 1% NaOH solution. 0.5 mL of sample solution was added into the mobile phase to be adjusted to 10 mL, and the sample was analyzed after passing through 0.22 μm microporous filter membrane. Peak area was quantified by external standard method, with benzoic acid serving as the standard (Fig. S2a ), and the TGP content was evaluated using the following formula, where 480.27 is the molecular weight of paeoniflorin and 122.12 is the molecular weight of benzoic acid.
7
HPLC Analysis of Compounds
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HPLC analysis was performed on an Agilent Technology HPLC 1260 infinity II. The chromatographic separation was accomplished with an InfinityLab Poroshell 120 EC-C18 column (4.6 mm × 150 mm, 4.0 μm) at 25°C. Two mobile were used including water containing 2% acetic acid (A) and acetonitrile (B). The isocratic elution was performed with a flow rate of 1 ml/min. The elution was set at 40% B for 30 min. Before analysis, the filtered (0.45 μm nylon membrane) mobile phases were degassed using an ultrasonic bath for 30 min. The injection volume was 10 μl. Detection wavelength was 425 nm.
8
HPLC and GC/MS Analytical Workflow
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High-performance liquid chromatography (HPLC 1260 Infinity II, Agilent, Santa Clara, CA, USA) analysis was carried out as described in [39 (link)] using isocratic separation with acetonitrile (20%) and water with 5mM ammonium formate and 0.1% formic acid (80%) at a constant flow rate of 1.5 mL/min. Gas chromatograph with mass selective detector (GC/MS 8860 GC/5977BMSD, Agilent, Santa Clara, CA) analysis was carried out also as described in [39 (link)].
9
HPLC-based HCQ Dissolution Profiling
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The Agilent HPLC 1260 Infinity-II was used for the estimation of HCQ dissolution profile. It consists of four channels, pressure range up to 600 bar, degasser with an integrated purge valve, thermostatic sampler, and column compartment. The PDA detector connected to empower 3 software (Build 3471 SPs Installed: Feature Release 3 DB ID: 2639633283) to monitor the output signal. The Disteck Premiere 5100 model dissolution apparatus was used to perform the multimedia profile. The column is Agilent Zorbax C8, 250 mm × 4.6 mm i.d., 5 μm. Sartorius analytical balances were used for the weighing of standards and samples. Bio-Technics ultra sonicator is used to extract the drug from the sample matrix.
10
Characterization of Ionic Liquid [BPy][BF4]
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The synthesized
ionic liquid product ([BPy][BF4]) was characterized by
NMR and IR analyses. The 1H-NMR and 13C-NMR
NMR spectra of [BPy][BF4] were recorded on a Bruker Avance
300 MHz spectrometer. Dimethyl sulfoxide (DMSO) was used as the solvent.
Oil-phase samples from EDS and OEDS processes were analyzed by
an Agilent Cary 6000i UV–vis–IR spectrophotometer at
a wavelength of λmax 284 nm to determine DBT concentration.
Standard solutions with DBT concentrations of 50, 30, 20, 15, 10,
7, and 5 ppm in n-hexane were used to get the calibration
curve.
An Agilent HPLC 1260 Infinity II was used for the quantitative
assay of DBT in the oil phase from the ODS test. The system was equipped
with a quaternary pump, an autosampler, a Zorbax XDB-C18 column (4.6
× 250 mm, 5 μm), and a diode array UV detector. The mobile
phase was 90% methanol in 10% water (v/v, %) with a flow rate of 1.0
mL/mm. The wavelength of UV was set at 284 nm. For the quantification
of DBT, the external standards with DBT contents of 500, 400, 300,
200, and 100 ppm were used to get the calibration curve.
ionic liquid product ([BPy][BF4]) was characterized by
NMR and IR analyses. The 1H-NMR and 13C-NMR
NMR spectra of [BPy][BF4] were recorded on a Bruker Avance
300 MHz spectrometer. Dimethyl sulfoxide (DMSO) was used as the solvent.
Oil-phase samples from EDS and OEDS processes were analyzed by
an Agilent Cary 6000i UV–vis–IR spectrophotometer at
a wavelength of λmax 284 nm to determine DBT concentration.
Standard solutions with DBT concentrations of 50, 30, 20, 15, 10,
7, and 5 ppm in n-hexane were used to get the calibration
curve.
An Agilent HPLC 1260 Infinity II was used for the quantitative
assay of DBT in the oil phase from the ODS test. The system was equipped
with a quaternary pump, an autosampler, a Zorbax XDB-C18 column (4.6
× 250 mm, 5 μm), and a diode array UV detector. The mobile
phase was 90% methanol in 10% water (v/v, %) with a flow rate of 1.0
mL/mm. The wavelength of UV was set at 284 nm. For the quantification
of DBT, the external standards with DBT contents of 500, 400, 300,
200, and 100 ppm were used to get the calibration curve.
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