G1312b binary pump

Manufactured by Agilent Technologies

Sourced in United States

The G1312B binary pump is a high-performance liquid chromatography (HPLC) pump designed to deliver precise and reproducible solvent flow rates. It features two independent solvent channels that can be combined to create binary mobile phases. The pump's core function is to provide a consistent and reliable flow of solvents for HPLC applications.

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G1312B binary pump SL (2 citations)

8 protocols using g1312b binary pump

HPLC-ESI-TOF-MS Analysis of GCE, eGCE and Tannic Acid

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GCE, eGCE and tannic acid were analyzed by high performance liquid chromatography coupled to electrospray ionization-time of flight-mass spectrometry (HPLC-ESI-TOF-MS) [19 (link)], a tandem system. The Agilent 1200 series HPLC consisted of a model G1315D diode array detector (DAD), a G1312B binary Pump, a G1379B vacuum degasser, a G1367C auto-sampler and a G1316B column heater. The concentration of the testing solution was 1mg/mL, and the injection volume was 5 μL. Gradient elution HPLC was applied at a flow rate of 0.4 mL/min with detection at 270 nm. Two solvents were used for the mobile phase: (A) 0.1% formic acid and 10 mM ammonium formate (pH3.0), (B) acetonitrile. Compounds were separated using the following gradient: 0-5 min, 5% B; 5-8 min, 20% B; 20-30 min, 30% B; 30-40 min, 20% B; 40.0-40.1 min, 5% B. The separation of components in GCE was performed on an Agilent column (Poroshell 120 SB-C18, 150mm × 4.6mm, particle size 2.7μm) at 40 °C.
The time of flight (TOF) mass detector (G1969A, Agilent, US) was equipped with electrospray ionization (ESI) interface. The ESI voltage was 3.5 kV, and a mass range of 50-3000 m/z was scanned in negative full scan mode. Data processing was performed on Agilent Mass Hunter (v.B.01.04) software.

Huang X., Deng M., Liu M., Cheng L., Exterkate R.A., Li J., Zhou X, & Ten Cate J.M. (2017). Comparison of Composition and Anticaries Effect of Galla Chinensis Extracts with Different Isolation Methods. The Open Dentistry Journal, 11, 447-459.

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HPLC-MS/MS Quantification of Analytes

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The specimens were analyzed using an Agilent Technologies 1200 HPLC system that consisted of G1367D autosampler, a G1379B degasser, a G1312B binary pump, and a G1310A isocratic pump (Wilmington, DE). Separation was achieved using a Synergi Hydro RP (50 mm × 2.0 with 2 μm particle size) C-18 column (Phenomenex, Torrence, CA). The column was held at 40°C in a G1316B thermostated column compartment (Wilmington, DE). The solvent system consisted of A (10 mm ammonium acetate/0.1% formic acid) and B (acetonitrile/0.1% formic acid) with a flow rate of 0.250 mL/min. The solvent program held B at 4.0% from 0.0 minutes to 7.0 minutes, increased to 40.0% from 7.0 minutes to 7.1 minutes, and decreased to 4% from 7.1 minutes to 11 minutes.
The detector was an AB Sciex 5500 mass spectrometer equipped with electrospray ionization in the positive mode. The ion spray voltage was set at 5500 V, and the source temperature was 650°C. The curtain gas and collision gas was nitrogen and was held at 30 psi and 5 psi, respectively. The transition parameters are listed in Table 1. All data were processed using Analyst 1.5.1 (Foster City, CA).

Jones J.T., Jones M., Jones B., Sulaiman K., Plate C, & Lewis D. (2015). Detection of Codeine, Morphine, 6-Monoacetylmorphine, and Meconin in Human Umbilical Cord Tissue: Method Validation and Evidence of In Utero Heroin Exposure. Therapeutic Drug Monitoring, 37(1), 45-52.

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HPLC Measurements of Drug Metabolites

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For HPLC measurements, an Agilent Technologies 1200 Series equipped with G1379B degasser, G1312B binary pump SL, G1367C HiP-ALS SL autosampler, a G1314C VWD SL UV detector, G1316B TCC SL column oven and a G1956B MSD mass selective detector was used. The analytes were separated on an Agilent Zorbax SB-C18 column (1.8 µm; 4.5 × 50 mm) at 50°C by using aqueous eluent (0.1% formic acid) and ACN at a flow of 1.0–1.2 mL min⁻¹ which was split to 0.6–0.8 mL min⁻¹ before the mass selective detector.
The following gradients were used to separate substrate and products from whole cell conversions: benzydamine (0–2.10 min: 20–70% ACN, 2.10–2.20 min: 70–90% ACN, 2.20–2.40: 90% ACN, 2.40–2.60 min: 90–20% ACN); ethionamide (0–1.20 min: 0–70% ACN, 1.20–2.10 min: 70% ACN, 2.10–3.00: 70–0% ACN); propranolol (0–3.00 min: 0–100% ACN, 3.00–3.50 min: 100% ACN, 3.50–4.00: 100–0% ACN); trifluoperazine (0–2.50 min: 15–100% ACN, 2.50–3.50 min: 100% ACN, 3.50–3.51: 100–15% ACN).

Geier M., Bachler T., Hanlon S.P., Eggimann F.K., Kittelmann M., Weber H., Lütz S., Wirz B, & Winkler M. (2015). Human FMO2-based microbial whole-cell catalysts for drug metabolite synthesis. Microbial Cell Factories, 14, 82.

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Comprehensive Analytical Techniques for Sample Characterization

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An LC series 1200 instrument (Agilent, Waldbronn, Germany) consisting of a model G1379B degasser, G1312B binary pump SL, G1367C autosampler SL, and G1316B column thermostat SL was employed for LC-hrMS/MS analysis. The LC was coupled online to an LTQ-Orbitrap XL high-resolution tandem mass spectrometer (Thermo Electron, Bremen, Germany) fitted with a heated electrospray ionization (HESI) source. For AAA by IDMS, a microwave hydrolysis system (CEM, Saclay, France) and a rotational vacuum concentrator connected to a freeze-dryer system (Christ, Germany) were used to hydrolyse and concentrate samples, respectively. An Exion LC coupled online to a QTRAP 6500+ mass spectrometer (AB Sciex, Les Ulis, France) fitted to an electrospray ionisation (ESI) source was used for the detection of amino acids. A 930 compact IC flex and 863 compact autosampler (Metrohm AG, Switzerland) and a Metrohm Titrando 851 coupled to a Metrohm 874 Oven sample processor were used for IC and KF titration, respectively. Elemental analysis was performed using a Vario Micro Cube Elemental Analyser (Elementar, Lyon, France), whereas NMR measurements were performed on an ECS-400 MHz NMR instrument equipped with a direct type (Royal) automatic tuning probe (JEOL, Tokyo, Japan).

Briones A., Martos G., Bedu M., Choteau T., Josephs R.D., Wielgosz R.I, & Ryadnov M.G. (2022). An SI-traceable reference material for virus-like particles. iScience, 25(5), 104294.

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Quantitative LC-MS/MS Analysis of Metabolites

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Liquid chromatography/quadrupole time-of-flight tandem mass spectrometry (LC/Q-TOF) analysis was performed on an Agilent 1260 LC system [consisting of a G1322A degasser, a G1312B binary pump, a G1367E thermostated autosampler, G1316A thermostated column compartment, and a G4212B diode array detector (DAD)] (Agilent Technologies, Santa Clara, CA, USA) coupled to an ultra high definition quadrupole time-of-flight mass spectrometer Model 6540 (Agilent Technologies, Santa Clara, CA, USA) equipped with a dual source electrospray ionization ion source. The sample treatment, LC separation and Q-TOF parameters were conducted according to previously described methods (Tang et al., 2015 (link)).
The analytes were separated on an Agilent C18 reverse-phase column (100 × 1.8 mm, particle size of 3.5 μm). The binary mobile phase composed of 2% methanol and 98% water (containing 0.2% ammonium acetate and 0.1% acetic acid) was set at a constant flow rate of 300 μL/min and the column temperature was kept at 30°C, and the sample volume was 2 μL. Q-TOF parameters were as previously reported (Tang et al., 2015 (link)).

Zheng C., Ma Y., Wang X., Xie Y., Ali M.K, & He J. (2015). Functional analysis of the sporulation-specific diadenylate cyclase CdaS in Bacillus thuringiensis. Frontiers in Microbiology, 6, 908.

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LCMS and HRMS Analysis of Compounds

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LC-MS includes Agilent 1100 series LC system consisting of G1312B binary pump, G1313A autosampler and 1200 Series diode-array detector (Agilent Technologies, Santa Clara, CA, USA) with an analytic column (Hypersil GOLD aQ, 3 µm C18, 150 x 2.1 mm) (Thermo Fisher Scientific) run on a gradient solvent of MeOH-H2O (50:50–100:0) at a flow rate of 0.2 mL/m. The eluted compounds were then analyzed with mass spectrometry using (+) ESI mode on the Agilent 6220 TOF mass spectrometer (Gas temperature - 350˚C, Drying gas (N2)–8 L/m. Nebulizer–2 Bar).
High-resolution mass spectrometry was recorded using (+) ESI mode on the Bruker Daltonics, Impact II QTOF mass spectrometer (Gas temperature −200 °C, Drying gas (N₂)–4 L/m Nebulizer–0.3 Bar, Bruker Scientific LLC, Billerica, MA, USA). Both LC-MS and high-resolution mass spectrometry analyses were conducted at the University of Florida Mass Spectrometry Research and Education Center, Department of Chemistry.

Niu G., Hao Y., Wang X., Gao J.M, & Li J. (2020). Fungal Metabolite Asperaculane B Inhibits Malaria Infection and Transmission. Molecules, 25(13), 3018.

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LC-MS/MS Quantification of Drugs

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The assay was developed and validated on an Agilent 1260 Infinity LC system, consisting of a G1312B binary pump, a G1367E autosampler, a G1330B thermostat, and a G1316A thermostatted column compartment, coupled with an Agilent 6430 triple-quadrupole mass spectrometer (Agilent, Santa Clara, CA, USA). The temperature of the autosampler was set at 4 °C. The chromatographic separation was performed on an Atlantis T3 column (2.1 × 100 mm, 3 μm; Waters Co., Milford, MA, USA), and temperature was set at 30 °C. The mobile phase was a gradient of 0.1% formic acid in water (solvent A) and 100% acetonitrile (solvent B), with a constant flow rate of 0.3 mL/min (Table 2). The total running time was 10.50 min for each injection.
Mass spectrometric analysis was carried out in positive ion mode and dynamic multiple reaction monitoring (MRM) with a gas temperature of 350 °C, a gas flow of 9 L/min, a nebulizer pressure of 30 psi and a positive capillary voltage of 2,000 V. Mass transitions and relevant mass spectrometer conditions for all drugs and internal standards are summarized in Table 1. Integration of peak height and peak area, as well as data analysis were performed using Agilent MassHunter software version B.06.00 (Agilent, Santa Clara, CA, USA).

Zheng X., Jongedijk E.M., Hu Y., Kuhlin J., Zheng R., Niward K., Paues J., Xu B., Davies Forsman L., Schön T., Bruchfeld J, & Alffenaar J.C. (2020). Development and validation of a simple LC-MS/MS method for simultaneous determination of moxifloxacin, levofloxacin, prothionamide, pyrazinamide and ethambutol in human plasma. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 1158.

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Analytical HPLC-QTOF Characterization

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Chromatographic measurements were performed with an Agilent Technologies (Santa Clara, CA, USA) 1200 Series instrument equipped with G1312B binary pump and G1367D autoinjector. In general, an UHD 6540 Accurate-Mass Q-TOF detector with electrospray ionization (ESI) was used for compound detection in most solutions.
However, a G1315C DAD was also used at 254 nm for detection in the phosphate buffers. Instrument control and processing were performed by Masshunter software 4.0.
A 100 mm, 4.6 mm i.d, 2.6 µm octadecylsilica Kinetex EVO C18 analytical column provided by Phenomenex (Torrance, CA, USA) with a core-shell Technology was used for all determinations. This material is stable within the pH range 1-12. pH measurements of mobile phase were done with a combined Crison 5202 electrode in a Crison 2001 pH meter (Hach Lange Spain, L'Hospitalet de Llobregat, Spain). The electrode system was calibrated with ordinary aqueous buffers of pH 4.01, 7.00 and 9.21 (25 ºC).

Soriano-Meseguer S., Fuguet E., Port A, & Rosés M. (2019). Influence of the acid-base ionization of drugs in their retention in reversed-phase liquid chromatography. Analytica chimica acta, 1078.

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