Overnight cultures of Bacillus amyrloliquefaciens were inoculated (5%, v/v) into 200 mL modified Landy medium, and then shaken at 33°C and 180 rpm for 36 h. At the end of cultivation, 200 mL supernatant was treated with 6 mol/L HCl to adjust the pH to 2.0 and then the antibacterial peptides was extracted with 5 mL methanol for different times and adjusted the pH to 7.0. The supernatants were analyzed by reversed-phase HPLC (C18 column, ODS 4.6×250 mm, AGILENT 1100 series). Total lipopeptides were determined by high-performance liquid chromatography (HPLC) (AGILENT 1100 series) using a C18 column (ODS 4.6×250 mm) with UV detectors. The eluent used was Methyl Cyanides with 1‰ trifluoroacetic acid at a flow rate of 0.6 mL/min. The injection volume of the sample was 20 µL. For fengycin production, Rhizopus stolonifer was used as an indicator. Glucose was estimated with DNS by measuring the OD (optical density) at 600 nm with Unic 721 spectrophotometer (Shanghai). Dry cell weight was determined after centrifuging 10 mL fermentation broth, washed twice with distilled water, and dried to constant weight at 85°C. The same sample was repeated three times.
1100 series
Manufactured by Agilent Technologies
Sourced in United States, Germany, Canada, Spain, Japan, Italy, France
The 1100 series is a line of analytical laboratory equipment manufactured by Agilent Technologies. The core function of the 1100 series is to perform liquid chromatography analysis.
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Lab products found in correlation
Eclipse xdb c18, by Agilent Technologies (6 mentions)
Hi plex h column, by Agilent Technologies (4 mentions)
Eclipse plus c18, by Agilent Technologies (3 mentions)
Chemstation, by Agilent Technologies (3 mentions)
Ascentis c18 column, by Merck Group (3 mentions)
Chemstation software, by Agilent Technologies (3 mentions)
Zorbax sb c18 column, by Agilent Technologies (3 mentions)
Whatman, by Cytiva (3 mentions)
Api 3000, by AB Sciex (3 mentions)
Advia centaur immunoassay system, by Siemens (3 mentions)
Luna c18 2 column, by Agilent Technologies (2 mentions)
Eclipse xdb c8 column, by Agilent Technologies (2 mentions)
1100 series hplc system, by Agilent Technologies (2 mentions)
Hplc 2d chemstation software, by Agilent Technologies (2 mentions)
Eclipse xdb c18 column, by Agilent Technologies (2 mentions)
Api6500 q trap, by AB Sciex (2 mentions)
380 lc series, by Agilent Technologies (2 mentions)
Zetasizer nano z, by Malvern Panalytical (2 mentions)
Uranyl acetate, by Electron Microscopy Sciences (2 mentions)
Zorbax eclipse xdb c18, by Agilent Technologies (2 mentions)
251 protocols using 1100 series
1
Optimizing Lipopeptide Production in Bacillus
2
HPLC Purification of Fluorescent Probes
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HPLC purification of probes was performed on an Agilent 1100 Series HPLC system (California, USA), consisting of a quaternary pump with solvent degasser, a diode-array module for multi-wavelength signal detection using an Agilent 1100 Series UV-visible detector and an Agilent 1100 Series fluorescence detector for on-line acquisition of excitation/ emission spectra. The system had a manual injector and thermostatted column compartment with two heat exchangers for solvent pre-heating. The HPLC system was operated by Agilent HPLC 2D ChemStation Software. Depending on the purification performed, the columns used were: Zorbax Eclipse X DB-C8 column (California, USA) (length 25 cm, inner diameter 4.6 mm, particle size 5μm), or a Luna C18 (2) column (California, USA) (length 25 cm, inner diameter 4.6 mm, particle size 5 μm) with elution using an increasing gradient (0-50%) of acetonitrile in water (fraction detection at 260, 280, and 340 nm).
3
HPLC Purification of Fluorescent Probes
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HPLC purification of probes was performed on an Agilent 1100 Series HPLC system (California, USA), consisting of a quaternary pump with solvent degasser, a diode-array module for multi-wavelength signal detection using an Agilent 1100 Series UV-visible detector and an Agilent 1100 Series fluorescence detector for on-line acquisition of excitation/emission spectra. The system had a manual injector and thermostatted column compartment with two heat exchangers for solvent pre-heating. The HPLC system was operated by Agilent HPLC 2D ChemStation Software. Depending on the purification performed, the columns used were: Zorbax Eclipse X DB-C8 column(California, USA) (length 25 cm, inner diameter 4.6 mm, particle size 5μm), or a Luna C18 (2) column (California, USA) (length 25 cm, inner diameter 4.6 mm, particle size 5 μm) with elution using an increasing gradient (0-50%) of acetonitrile in water (fraction detection at 260, 280, and 340 nm).
4
High-resolution HPLC analysis of phenolic profiles
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The phenolic profile was determined using a high-resolution liquid chromatography system, HPLC (Hewlett-Packard 1100 series equipped with an array diode detector and an Agilent 1100 series automatic injector which introduces 20 µL of sample). The chromatographic column used was Teknokroma Tracer Extrasil OSD2 of 5 µm particle size and dimensions of 25 × 0.46 internal diameter. As an eluent, HPLC grade acetonitrile (B) and mill-Q water with 0.01% in Trifluoroacetic acid (TFA) were used. The flow rate was 1 mL/min and the chromatograms were recorded at 254, 280, and 340 nm. Phenolic compounds were separated using the following gradient: 0–30 min, 5% B; 30–45 min, 25% B; 45–47 min, 50% B; 47–50 min, 0% B.
The identification and quantification of phenolic compounds were based on the comparison of the retention times (RT) and absorbance values of detected peaks in solvents with those obtained by the injection of pure standards of each analysis.
The identification and quantification of phenolic compounds were based on the comparison of the retention times (RT) and absorbance values of detected peaks in solvents with those obtained by the injection of pure standards of each analysis.
5
HPLC Analysis of Amino Acids
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HPLC analysis of amino acids was performed with an Agilent 1100 series equipped with a diode array detector (DAD) and a ZORBAX Eclipse XDB-C18 column, 4.6 mm ID × 250 mm (5 µm) 80 Å. This setup was used with the Agilent protocol for HPLC analysis of amino acids98 .
6
Quantitative HPLC Analysis of Curcumin
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Curcumin and curcuminoids were prepared with 100% methanol and quantitated with an HPLC system (Agilent 1100 series, Germany) equipped with a Zorbax Eclipse C18 column (250 × 4.0 mm2). The mobile phase consisted of 1% acetic acid in water (A) and 52% acetonitrile (B); the column was equilibrated for 10 min in the mobile phase and then washed with 100% (B) for 10 min. The column was operated at room temperature with a 1 ml/min flow rate. The injection volume was 5 μl, and curcumin, curcuminoids, bis-demethoxycurcumin (BDMC), and demethoxycurcumin (DMC) were detected at a wavelength of 424 nm.
7
Quantification of Capsaicin in Chili Peppers
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The identification and quantification of CAP were performed according to the methodology of Othman et al. (20 (link)). In brief, after extraction from CPSs, the supernatant was filtered through a 0.45 μm nylon filter into an HPLC sample vial using a disposable syringe. The chromatographic analysis of CAP and dihydrocapsaicin (DHC) was performed using high-performance liquid chromatography (HPLC) (Agilent 1100 Series Santa Clara, CA, United States) with a Zorbax Eclipse Plus C18 (particle size 4.6 × 100 mm 3.5 μm) column at 40°C, a flow of 0.8 ml/min, and an injection volume of 10 μl. The mobile phase consisted of (A) HPLC-grade water with 0.1% of formic acid and (B) HPLC-grade methanol. The gradient started with 50% of B for 2 min, increasing to 70% in 5 min, and in the next 5 min, the percentage of B changed to 80%. After 15 min, the percentage of B increased to 100% and remained in an isocratic mode for another 5 min. Chromatograms were obtained at 280 nm. The standard curve for CAP was used (y = 4.9188x + 5.7049; R2 = 0.9999). The CAP and DHC concentrations in samples were expressed as μg of CAP equivalents/ml of extract.
8
HPLC Analysis of Amino Acids
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HPLC analysis of amino acids was performed with an Agilent 1100 series equipped with a diode array detector (DAD) and a ZORBAX Eclipse XDB-C18 column, 4.6 mm ID x 250 mm (5 µm) 80 Å. This setup was used with the Agilent protocol for HPLC analysis of amino acids [108] .
9
Serum Nucleotide Quantification by RP-HPLC
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Mice serum was extracted with 1.3 M perchloric acid (Sigma-Aldrich, Burlington, MA, USA) (1:1 ratio). Levels of nucleotides were measured by a reverse-phase high-pressure liquid chromatography (RP-HPLC) method using the liquid chromatography (LC) system (Agilent Technologies 1100 series, Agilent Technologies Inc., Santa Clara, CA, USA), as described previously [15 (link),30 (link)]. Results are presented as μmol/L.
10
Reversed-Phase HPLC-MS/MS Peptide Analysis
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After the digestion, the tryptic peptides were dissolved in 100 μl of 0.1% (v/v) trifluoroacetic acid and subjected to a reversed phase chromatography (C18 Gemini-NX, μl particle size, 110 Å pore size, 250x4.6 mm, Phenomenex, Torrance, CA.) connected to a HPLC Agilent Technologies 1100 Series (Agilent Technologies, Santa Clara, CA.). The column effluent was analyzed by MS using an electrospray ion trap mass spectrometer (Agilent Technologies LC/MSD Trap SL) operating in positive ion mode over the mass range 300-2200 amu (atomic mass units). MS spray voltage was 3.5 kV and the capillary temperature was maintained at 300°C. Obtained spectra were extracted and analyzed by the MASCOT software (www.matrixscience.com) and by the SONAR software (http://hs2.proteome.ca/prowl/knexus.html) with the following search parameters: database, NCBInr; taxonomy, Eukaryota; enzyme, trypsin; peptide tolerance, 1.2 Da; MS/MS tolerance, 0.6 Da and allowance of one missed cleavage [26 (link)].
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