Reverse phase HPLC (RP-HPLC) analysis was performed on an Agilent Eclipse XDB-C18 (4.6 × 150 mm, 3.5 μm) column using an Agilent 1200 liquid chromatograph (Omaha, Nebraska, USA). For the analysis of crude peptides (20 μL, 1 mg/mL), a linear gradient was applied from 5% to 70% Solvent B (0.05% TFA in ACN) in Solvent A (0.05% TFA in water) for 45 min at a flow rate of 1.0 mL/min at RT and 210 nm detection. The crude products were purified through solid-phase extraction (SPE), using Supelclean LC-18 SPE columns that were activated and equilibrated prior to use. Crude peptides were passed through the column, and a gradient was used for their elution [29 ]. Collected fractions were analyzed using RP-HPLC (as describe above) and MS. MALDI-TOF MS analysis was performed on an Ultraflex III TOF-TOF mass spectrometer (Bruker Daltonics, Bremen, Germany) in reflectron mode, using an MTP384 polished steel target (Bruker Daltonics), 2,5-dihydroxybenzoic acid, or sinapinic acid as a matrix, 500 shots with 25–30% power laser.
1200 liquid chromatograph
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1200 liquid chromatograph is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative applications. It provides precise and reliable separation and detection of a wide range of chemical compounds. The 1200 liquid chromatograph features a modular design, allowing for customization to meet specific analytical requirements.
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Lab products found in correlation
Api 5000 triple quadrupole mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Luna c18 column 2 x 50 mm, by Phenomenex (2 mentions)
Ultraflex 3 tof tof mass spectrometer, by Bruker (1 mentions)
Mtp384 polished steel target, by Bruker (1 mentions)
Eclipse xdb c18, by Agilent Technologies (1 mentions)
6410b triple quadrupole mass spectrometer, by Agilent Technologies (1 mentions)
Masshunter software, by Agilent Technologies (1 mentions)
6410 triple quadrupole mass spectrometer, by Agilent Technologies (1 mentions)
Masshunter software package, by Agilent Technologies (1 mentions)
Hypersil gold column, by Thermo Fisher Scientific (1 mentions)
Shim pack c18 column, by Shimadzu (1 mentions)
6410b triple quad mass spectrometer, by Agilent Technologies (1 mentions)
Zorbax eclipse plus c18 column, by Agilent Technologies (1 mentions)
Nexus 870 ftir spectrometer, by Thermo Fisher Scientific (1 mentions)
Methanol, by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Nitric acid, by Merck Group (1 mentions)
Zorbax sb c18 column, by Agilent Technologies (1 mentions)
Masshunter analysis software, by Agilent Technologies (1 mentions)
Guard column hardware kit, by Agilent Technologies (1 mentions)
10 protocols using 1200 liquid chromatograph
1
Purification and Characterization of Crude Peptides
2
Bilirubin Quantification in Serum and Colon
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The bilirubin levels in serum and colon homogenates were measured by ion-pair extraction using 0.1 mol/L methanolic di-n-octylamine acetate. Briefly, the supernatants were injected into the liquid chromatography mass spectrometer (LC-MS/MS) system was equipped with an Agilent 1200 liquid chromatograph and a 6410B triple quadrupole mass spectrometer with an ESI source. Data were analyzed using the MassHunter software (Agilent Corporation, Lexington, MA, United States). Chromatography was performed on a Poroshell 120 EC-C18 2.1 μm 30 × 50 mm column (Agilent).
3
LCMS Analysis of NBBS Compound
4
Honey Metabolite Profiling by LC-MS/MS
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The honey extracts were subjected to liquid chromatography/tandem mass spectrometry (LC-ESI-MS/MS). The system was an Agilent 1200 liquid chromatograph and a 6410 triple quadrupole mass spectrometer with an electrospray ionization source. Data were analyzed using the MassHunter software package (Agilent Corporation, MA, and USA). Separation was performed on a Zorbax C18 column (2.1 mm × 50 mm, 0.18 μm particle size) by gradient elution using the following gradient: 0–1 min 5% B isocratic; 1–14 min linear gradient to 100% B; 14–15 min to 5% B and the column was reconditioned at 5% B for 5 min. Eluent A was water with 0.1% formic acid and B was methanol with 0.1% formic acid. The acid used in the analysis improves the chromatographic peak shape and provides a source of proton in reverse phase LC-MS. The column temperature was 30° C. Mass spectrometry was performed on triple quad spectrometer in ESI positive ion mode. High purity nitrogen served as both the nebulising agent and the drying gas, with a drying gas flow 9 l/min and a nebuliser pressure 25 psi at 350°C. The capillary voltage was set at 4,000 V and the source at 300°C.
5
Organic Acid Quantification by HPLC
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The organic acids produced by the strain tested were identified and quantified using high-performance liquid chromatography (Agilent 1200 liquid chromatograph, Santa Clara, USA) after the strain was cultured in 50 ml NBRIP liquid medium on a shaker at 25°C for 4–5 days (Xi et al., 2007 ).
Gluconic acid was measured using an Agilent ZORBAX SB-Phenyl chromatography column (4.6×250 mm, 5 μm). The mobile phase consisted of (A) 0.1% H3PO4 and (B) CH3CN at a flow rate of 1 ml/min. The gradient was 40% B phase at 0–4 min and 90% B phase at 5–7 min. The injection volume was 2 μl and the determination wavelength 203 nm (Zeng et al., 2016 (link)). Other organic acids were measured in a Thermo Hypersil Gold column (250×4.6 mm, 5 μm); mobile phase, (A) 50 mmol/l KH2PO4, pH 2.5 and (B) methanol; flow rate, 1 ml/min; gradient, 0–8 min 0% B phase, 8–13 min 40% B phase; determination wavelength 203 nm; injection volume 2 μl.
Gluconic acid was measured using an Agilent ZORBAX SB-Phenyl chromatography column (4.6×250 mm, 5 μm). The mobile phase consisted of (A) 0.1% H3PO4 and (B) CH3CN at a flow rate of 1 ml/min. The gradient was 40% B phase at 0–4 min and 90% B phase at 5–7 min. The injection volume was 2 μl and the determination wavelength 203 nm (Zeng et al., 2016 (link)). Other organic acids were measured in a Thermo Hypersil Gold column (250×4.6 mm, 5 μm); mobile phase, (A) 50 mmol/l KH2PO4, pH 2.5 and (B) methanol; flow rate, 1 ml/min; gradient, 0–8 min 0% B phase, 8–13 min 40% B phase; determination wavelength 203 nm; injection volume 2 μl.
6
LCMS Analysis of NBBS Compound
7
HPLC-MS Analysis of Ginsenosides
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The amount of Rb1, Rg1 and R1 was analyzed by an HPLC system. A Shimadzu shim-pack C18 column (250 mm × 4.6 mm, 5 μm) was used as the stationary phase. The mobile phase consisted of ultra-water (A) and acetonitrile (B) and was run in an isocratic mode at a flow rate of 1.0 mL·min−1. The following gradient was performed: 0–23 min, 23% B; 23–40 min, 35% B. The column temperature was maintained at 35 °C. Aliquots of 20 μL were injected. Monitoring and quantitation of Rb1, Rg1 and R1 were performed at 204 nm.
Agilent 1200 liquid chromatograph combined with an Agilent 6410B Triple Quad mass spectrometer was used to identify Rb1, Rg1 and R1 in biological samples. An Agilent zorbax eclipse plus C18 column (150 mm × 2.1 mm, 5 μm) was used as the stationary phase. The mobile phase for LC-MS analysis consisted of acetonitrile-water (5:5, v/v) at a flow rate of 0.5 mL·min−1.
Agilent 1200 liquid chromatograph combined with an Agilent 6410B Triple Quad mass spectrometer was used to identify Rb1, Rg1 and R1 in biological samples. An Agilent zorbax eclipse plus C18 column (150 mm × 2.1 mm, 5 μm) was used as the stationary phase. The mobile phase for LC-MS analysis consisted of acetonitrile-water (5:5, v/v) at a flow rate of 0.5 mL·min−1.
8
Flavonoid Analysis of Citrus limetta
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Citrus limetta was collected from local gardens. Rutin with HPLC grade, methanol, nitric acid, and acetic acid was purchased from Sigma‐Aldrich Inc., and graphene oxide nanoplatelets (99%, Thickness 3.4–7 nm with 6–10 Layers) were used as received. Fourier transform infrared spectroscopy (FTIR) was recorded using KBr tablets on a Thermo Nicolet Nexus 870 FTIR spectrometer. Scanning electron microscope (SEM) was taken using an MIRA3\\TESCAN‐XMU model. The samples were studied by thermogravimetric analysis (TGA) (Netzsch TG 209 F1 Iris1) under nitrogen gas atmosphere (10℃/min). The concentration of flavonoids was measured by variable‐wavelength UV‐Vis spectrophotometry of Unico UV 2100 Model. High‐performance liquid chromatography (HPLC) was performed with an Agilent Technologies 1200 liquid chromatograph which equipped to a quaternary solvent‐delivery system (an auto sampler and a UV detector). An Agilent Zorbax SB‐C18 column (100 × 4.6 mm, 5 μm particle size) was used for all analyses. An ultrasonic bath with temperature control (Elmasonic, S60H) was used to disperse the GO in solution.
9
Amino Acid Quantification by LC-MS
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Media samples were analyzed by LC-MS based on the method of Liu et al. (50 ). A 6-point calibration curve was prepared using a mix of unlabeled amino acids (Sigma part AAS18 supplemented with L-asparagine, L-glutamine, and L-tryptophan) with the same internal standard-to-solvent ratio as for the samples. LC-MS used an Agilent 1200 liquid chromatograph coupled to an Agilent 6410 tandem quadrupole mass spectrometer. Amino acid concentration was quantified using Agilent MassHunter Analysis software, measuring peak area ratio to the matching calibrated internal standard.
10
Phenolic Profile of B. prostrata Leaves
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The object of the study is the leaves of the plants of B. prostrata in the flowering phase collected in the period 2019-2020 in Novosibirskaja Oblast', the Republics of Khakassia, Buryatia, Altai, Tyva, etc. (Table 1). The raw materials were dried in well-ventilated rooms, crushed to a particle size of 2 mm, mixed and an average sample was taken.
Double extraction with 70% ethanol in a water bath (60-70°C for 1.0-1.5 h.) was carried out to extract the phenolic compounds. An exact weight (0.1 g) of the crushed material was extracted with 30 ml of aqueous ethanol for 30 min followed by 20 ml for 20 min. The residue on the filter was washed with 5 ml of 70% ethanol. The combined extract was concentrated to 10-15 ml (exact volume). To free from impurities, 1 ml of the extract was diluted with bidistilled water to 5 ml and passed through a Diapack C16 cartridge.
The component composition of the phenolic complex of the samples was studied by high-performance liquid chromatography on an analytical HPLC system consisting of an Agilent 1200 liquid chromatograph. The chromatographic separation was conducted at 25°C on a Zorbax SB-C18 Column (4.6 × 150 mm, 5 µm internal diameter) with the Agilent Guard Column Hardware Kit. Detection was performed at a wavelength of λ = 360 nm [11] . Quantification of individual components in plant samples was carried out using the external standard method.
Double extraction with 70% ethanol in a water bath (60-70°C for 1.0-1.5 h.) was carried out to extract the phenolic compounds. An exact weight (0.1 g) of the crushed material was extracted with 30 ml of aqueous ethanol for 30 min followed by 20 ml for 20 min. The residue on the filter was washed with 5 ml of 70% ethanol. The combined extract was concentrated to 10-15 ml (exact volume). To free from impurities, 1 ml of the extract was diluted with bidistilled water to 5 ml and passed through a Diapack C16 cartridge.
The component composition of the phenolic complex of the samples was studied by high-performance liquid chromatography on an analytical HPLC system consisting of an Agilent 1200 liquid chromatograph. The chromatographic separation was conducted at 25°C on a Zorbax SB-C18 Column (4.6 × 150 mm, 5 µm internal diameter) with the Agilent Guard Column Hardware Kit. Detection was performed at a wavelength of λ = 360 nm [11] . Quantification of individual components in plant samples was carried out using the external standard method.
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