1H and NMR spectral data were recorded in CDCl3 or DMSO-d6 on a 300 or 400 MHz Bruker NMR spectrometer. Column chromatography was conducted on Revelaris flash chromatography system. Reactions were monitored using thin-layer chromatography (TLC) on silica gel plates. HPLC analysis was conducted on an Agilent 1100 series LC system (Agilent ChemStation Rev.A.10.02; Phenomenex-Luna-C18, 4.8 mm × 150 mm, 5 μm, 1.0 mL/min, UV 254nm, room temperature) with MeCN/H2O (0.05% TFA or HCOOH buffer) gradient elution. HPLC-MS was performed on a Gilson 321 HPLC with detection performed by a Gilson 170 DAD and a Finnigan AQA mass spectrometer operating in electrospray ionization mode using a Phenomenex Gemini C18 150x4.6mm column. Compound purity was determined using an Agilent 1100 series LC system (Agilent ChemStation Rev.A.10.02; Phenomenex-Luna-C18, 4.8 mm × 150 mm, 5 μm, 1.0 mL/min, UV 254nm, room temperature) with MeCN/H2O (0.05% TFA or HCOOH buffer) gradient elution. All compounds were >95% pure via LC/MS analysis.
Agilent 1100 series lc system
Manufactured by Agilent Technologies
Sourced in United States, Germany
The Agilent 1100 series LC system is a high-performance liquid chromatography (HPLC) system designed for analytical applications. It features a modular design with various components such as a solvent delivery system, autosampler, column compartment, and detector. The system is capable of performing gradient and isocratic separations, providing reliable and reproducible results for a wide range of analytical tasks.
Automatically generated - may contain errors
Lab products found in correlation
Nmr spectrometer, by Bruker (2 mentions)
321 hplc, by Gilson (2 mentions)
Aqa mass spectrometer, by Phenomenex (2 mentions)
Xcalibur software, by Thermo Fisher Scientific (2 mentions)
Ltq ft ultra mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Gemini c18 150x4.6mm column, by Phenomenex (1 mentions)
Hystar version 3, by Bruker (1 mentions)
Esquire 6000 esi ion trap mass spectrometer, by Bruker (1 mentions)
Hp 6890 plus gc, by Hewlett-Packard (1 mentions)
Hp 5973 msd mass selective detector, by Hewlett-Packard (1 mentions)
Ammonium acetate, by Merck Group (1 mentions)
Phosphoric acid, by Merck Group (1 mentions)
Toluene, by Merck Group (1 mentions)
Ethylbenzene, by Merck Group (1 mentions)
Uracil, by Merck Group (1 mentions)
Sildenafil, by Merck Group (1 mentions)
Histidine, by Merck Group (1 mentions)
Propylparaben, by Merck Group (1 mentions)
Methanol meoh, by Merck Group (1 mentions)
Agilent 6410 triple quadrupole mass spectrometer, by Agilent Technologies (1 mentions)
24 protocols using agilent 1100 series lc system
1
Spectroscopic and Chromatographic Analysis
2
O-Methylflavonoid Analysis in E. coli
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The O-methylflavonoid content of E. coli culture extracts was analyzed using an Agilent 1100 Series LC system (Agilent Technologies) coupled to an ultraviolet diode array detector (UV-DAD, Agilent Technologies) and an Esquire 6000 ESI-Ion trap mass spectrometer (Bruker Daltonics). Chromatographic separation was performed on an EC 250/4.6 Nucleodur Sphinx column (RP 5 μm, Macherey-Nagel, Düren, Germany), with 0.2% (v/v) formic acid in water (A) and acetonitrile (B) as mobile phases. The flow rate was 1 mL/min and the column temperature was set to 25°C. The following elution profile was used: 0–15 min, 30–60% B; 15.1–16 min, 100% B; 16.1–20 min, 30% B. The mass spectrometer was run in alternating ion polarity (positive/negative) mode with a skimmer voltage of +40 V/−40 V, a capillary voltage of −3,500 V/+3,000 V and a capillary exit voltage of 113.5 V/−113.5 V, to scan masses from m/z 50–3,000. N2 was used as drying gas (11 L/min, 330°C) and nebulizer gas (35 psi). The software programs esquireControl version 6.1 (Bruker Daltonics) and HyStar version 3.2 (Bruker Daltonics) were used for data acquisition, while DataAnalysis version 3.4 was used for data processing. The UV absorption of individual O-methylflavonoids was analyzed using the post-processing software included in the HyStar version 3.2 package (Bruker Daltonics).
3
LC-MS and GC/MS Analysis of FCME
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LC-MS analysis of FCME was accomplished by Agilent 1100 series LC system (Agilent Technologies, Santa Clara, CA) equipped with an Agilent 1946B mass selective detector (MSD) (Agilent Technologies, Santa Clara, CA). A Pursuit XRs C18 (2.0 mm × 150 mm, 3 μm) column was utilized at a flow rate of 0.2 mL/min at 40°C. The mobile phase consists of A (0.1% formic acid in water) and B (0.1% formic acid in acetonitrile). The run time for each sample was 60 min, and the mobile phase started with 95% “A” and 5% “B” in the first 2 min; then “A” was decreased gradually to 0% until the 50th min and held for an additional 5 min. Thereafter, the proportion of “A” was increased to 95% within 5 min. The compounds were analyzed in selected ion monitoring (SIM) mode.
GC/MS analysis of FCME was performed by Kyungpook National University Center for Scientific Instruments and carried out using a HP 6890 Plus GC gas chromatograph with a (MSD)—HP 5973 MSD mass selective detector (Hewlett-Packard). Samples were diluted 1 : 1000 (v : v) with HPLC grade dichloromethane. Aliquots of the sample (1 μL) were injected into an HP-5 column. The GC oven temperature was set at 50°C for 4 min, increased to 280°C at a rate of 4°C/min, and held at the final temperature for 2 min. Velocity of the He carrier gas (99.99%) was 0.7 mL/min. Quantitative analysis was performed using the area normalization method.
GC/MS analysis of FCME was performed by Kyungpook National University Center for Scientific Instruments and carried out using a HP 6890 Plus GC gas chromatograph with a (MSD)—HP 5973 MSD mass selective detector (Hewlett-Packard). Samples were diluted 1 : 1000 (v : v) with HPLC grade dichloromethane. Aliquots of the sample (1 μL) were injected into an HP-5 column. The GC oven temperature was set at 50°C for 4 min, increased to 280°C at a rate of 4°C/min, and held at the final temperature for 2 min. Velocity of the He carrier gas (99.99%) was 0.7 mL/min. Quantitative analysis was performed using the area normalization method.
4
Phenol Degradation by Bacterial Cultures
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For phenol degradation experiments, the cells were harvested by centrifugation (10,000 g, 10 min) from 10 ml of preculture in Gause's synthetic medium at 25°C, 150 rpm for 72 h, then washed three times with sterile saline. The resulting inoculum was incubated in 250-ml Erlenmeyer flasks containing 100 ml of MSM supplemented with 3.0 mM phenol as the sole carbon source and incubated at 25°C, 150 rpm for 72 h in the dark. One milliliter per 100 ml liquid MSM in an Erlenmeyer flask was collected and passed through a 0.22 μm organic phase filter every 24 h, and then, 20 μl of the supernatant was analyzed. The assays were carried out in triplicate.
The Agilent 1100 Series LC system (Agilent, USA) controlled by the Agilent Chem Station Software and fitted with an Alltech ApolloC18 column (250 mm × 4.6 mm ID) was used to detect the concentrations of phenol and metabolites. The operating conditions were as follows: room temperature; mobile phase, deionized water/acetonitrile (70:30, v/v) with a solvent flow rate of 1.0 ml/min. The ultraviolet analysis was carried out at 270 nm.
The Agilent 1100 Series LC system (Agilent, USA) controlled by the Agilent Chem Station Software and fitted with an Alltech ApolloC18 column (250 mm × 4.6 mm ID) was used to detect the concentrations of phenol and metabolites. The operating conditions were as follows: room temperature; mobile phase, deionized water/acetonitrile (70:30, v/v) with a solvent flow rate of 1.0 ml/min. The ultraviolet analysis was carried out at 270 nm.
5
Analytical HPLC Method Development
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Methanol (MeOH) and acetonitrile
(AcCN) were of HPLC (gradient) grade and purchased from Sigma-Aldrich
(Germany). The studied compounds (uracil, benzene, toluene, ethylbenzene,
propylbenzene, propylparaben, butylparaben, sildenafil, and histidine)
were of proanalysis grade and purchased from Merck (Germany). The
additives (phosphoric acid and ammonium acetate) used for specific
aqueous components from the mobile phase were of proanalysis grade
purchased from Merck (Germany). Water of HPLC purity was obtained
within the laboratory with a TKA Lab HP 6UV/UF instrument (Thermo
Scientific). Solutions with a 100 μg/mL concentration of the
studied compounds were made in acetonitrile.
The HPLC experiments
were performed using an Agilent 1100 Series LC system (Agilent Technologies)
consisting of the following modules: a degasser (G1379A), binary pump
(G1312A), autosampler (G1313A), column thermostat (G1316A), and diode
array detector (G1315A). Chromatographic data were acquired by means
of Agilent Chemstation software rev. B.01.03.
Several columns
were used in this study, and some of their characteristics are described
inTable 1 . All columns
that have a C18 stationary phase are endcapped, and they were new
and tested before usage according to the manufacturer’s specifications.
(AcCN) were of HPLC (gradient) grade and purchased from Sigma-Aldrich
(Germany). The studied compounds (uracil, benzene, toluene, ethylbenzene,
propylbenzene, propylparaben, butylparaben, sildenafil, and histidine)
were of proanalysis grade and purchased from Merck (Germany). The
additives (phosphoric acid and ammonium acetate) used for specific
aqueous components from the mobile phase were of proanalysis grade
purchased from Merck (Germany). Water of HPLC purity was obtained
within the laboratory with a TKA Lab HP 6UV/UF instrument (Thermo
Scientific). Solutions with a 100 μg/mL concentration of the
studied compounds were made in acetonitrile.
The HPLC experiments
were performed using an Agilent 1100 Series LC system (Agilent Technologies)
consisting of the following modules: a degasser (G1379A), binary pump
(G1312A), autosampler (G1313A), column thermostat (G1316A), and diode
array detector (G1315A). Chromatographic data were acquired by means
of Agilent Chemstation software rev. B.01.03.
Several columns
were used in this study, and some of their characteristics are described
in
that have a C18 stationary phase are endcapped, and they were new
and tested before usage according to the manufacturer’s specifications.
6
Pectin Degradation Analysis by LC-ESI-MS
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The obtained products of pectin degradation were analyzed and identified by LC–ESI–MS. A 2 mg of each sample was dissolved in 100 μL distilled water and the soluble fraction of the samples was investigated. The investigation was performed isocratically by a mixture of water and acetonitrile (90:30) as a mobile phase and the flow rate was set to 0.3 mL/min. An Agilent 1100 series LC system consisting of a quaternary delivery pump, a thermostated column compartment, a degasser (Agilent Technologies, Germany), and a Rheodyne 7725i manual injector valve with a 20 µL sample loop (Cotati, CA, USA) was used to prepare a sample and the mass spectrometry was carried out with Agilent 6410 Triple Quadrupole mass spectrometer (Agilent Technologies, Palo Alto, CA, USA) which was run by Agilent MassHunter Workstation B.01.03. Ionization was attained using electrospray ionization (ESI) in the negative mode with the capillary voltage of 4000 V. Nitrogen was used as nebulizer gas with a nebulizer pressure of 40 psi and source temperature of 100 °C. Nitrogen was heated to 300 °C and delivered at a flow rate of 10 L/min. The fragment voltage for the samples was 280 V and the dwell time was 200 ms. The analyte was detected using scan mode32 (link).
7
HPLC-MS Analysis of 5-FU and 5-BU
8
Quantification of Berberine and Palmatine
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Frozen sample blocks were cut from bark to xylem into serial tangential sections of 100-µm thickness. Every two serial sections were collected in the same plastic tube (1.5 mL in volume) and extracted with 1 mL ACN at 60°C for one hour. HPLC measurements for berberine and palmatine were achieved with Agilent 1100 series LC system (Agilent Technologies Inc.) equipped with a TSKgel ODS-100S column (4.6 mm i.d. × 25 cm, C18, 5.0 μm, Tosoh Corp., Japan). Berberine and palmatine were separated by a binary buffer system of 0.1% (v/v) FA (buffer A) and 0.1% (v/v) FA in ACN (buffer B) at a flow rate of 1 mL/min. The gradient was 35 min in total and set as follows: from 5 to 20% (v/v) buffer B in 5 min, from 20 to 30% (v/v) buffer B in 15 min, from 30 to 95% (v/v) buffer B in 0.1 min, holding at 95% (v/v) buffer B for 9.9 min, declining to 5% (v/v) buffer B in 0.1 min, and holding at 5% (v/v) buffer B for 4.9 min. All the chromatograms were taken at column temperature 30°C and UV detection wavelength 265 nm. The measurements were carried out in three replicates using three blocks obtained from the same sample disk to evaluate the average amount and standard errors of berberine and palmatine.
9
LC-MS Analysis of Adenosine and Inosine
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The rBmpD and LF-rBmpD samples were heated and centrifuged, and the supernatant was directly used for LC-MS analysis with an Agilent 1100 series LC system. The analytical method was modified from the method described by Ren and colleagues (42 (link)). Separations were conducted using gradient elution on a SunFire C18 analytical column (2.1 by 150 mm; particle size, 3.5 μm; Waters, Milford, MA, USA). Mobile phases were 0.1% formic acid in water (solvent A) and 0.1% formic acid in methanol (solvent B). The gradient conditions were 5% solvent B (0 to 12 min), from 5% to 80% solvent B (12 to 13 min), 80% solvent B (13 to 18 min), from 80% to 5% solvent B (18 to 18.5 min), and 5% solvent B (18.5 to 25 min). The flow rate was 0.25 ml/min. Retention times for adenosine and inosine were 4.7 and 7.7 min, respectively (Fig. 4A ).
MS detection was performed in selected-ion monitoring mode with a single quadrupole mass spectrometer (HP 1100 LC/MSD). Ionization was based on electrospray ionization in positive-ion mode. The capillary voltage was 4.0 kV, and the drying gas temperature was 350°C. The selected ions for adenosine and inosine were m/z 268.0 and 269.0, respectively (Fig. 4A ). These masses correspond to the protonated molecules, [M+H]+. Adenosine was also detected as m/z 269.0, due to its isotopic distribution.
MS detection was performed in selected-ion monitoring mode with a single quadrupole mass spectrometer (HP 1100 LC/MSD). Ionization was based on electrospray ionization in positive-ion mode. The capillary voltage was 4.0 kV, and the drying gas temperature was 350°C. The selected ions for adenosine and inosine were m/z 268.0 and 269.0, respectively (
10
Quantitative Analysis of Piperacillin and Tazobactam
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Concentrations of piperacillin and tazobactam were analyzed using validated liquid chromatography–tandem mass spectrometry (LC–MS/MS), Agilent 1,100 series LC system (Agilent, Wilmington, DE, USA). Piperacillin, tazobactam, and their internal standard (doripenem) were prepared by protein precipitation with 60% acetonitrile. Detection and quantification were performed using a triple quadrupole tandem mass spectrometer with an electrospray ionization interface in the positive and multiple-reaction-monitoring modes. Chromatographic separation of the compounds was accomplished using an Atlantis HILIC column (50 × 2.1 mm, 3 µm; Waters, Milford, MA, USA) with an isocratic elution.
A full validation of the assay was carried out with respect to selectivity, accuracy, precision, recovery, calibration curve, and stability. The calibration curves for piperacillin and tazobactam were linear over ranges of 2.0–500.0 µg/mL and 0.2–50.0 µg/mL, respectively, with coefficients of determination (R2) greater than 0.995 for all.
A full validation of the assay was carried out with respect to selectivity, accuracy, precision, recovery, calibration curve, and stability. The calibration curves for piperacillin and tazobactam were linear over ranges of 2.0–500.0 µg/mL and 0.2–50.0 µg/mL, respectively, with coefficients of determination (R2) greater than 0.995 for all.
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