1260 hplc

Manufactured by Agilent Technologies

Sourced in United States, Germany

The Agilent 1260 HPLC is a high-performance liquid chromatography system designed for analytical separations and quantitative analysis. It features a modular design, allowing for customization to meet specific application requirements. The system is capable of delivering precise and reproducible mobile phase flow, ensuring reliable performance and data integrity.

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Agilent 1260 HPLC (156 citations) Agilent 1260 series HPLC system (82 citations) Agilent 1260 HPLC/6520 QTOF-MS instrument (4 citations) 1260 Infinity HPLC instrument (4 citations) 1260 In nity HPLC system (3 citations) 1260 Infinity Quaternary HPLC system (3 citations) Agilent 1260 Infinity Quaternary HPLC system (3 citations) Agilent 1260 Infinity HPLC-DAD (2 citations) Agilent 1260 Infinity HPLC chromatograph (2 citations) Agilent/1260-Infinity-preparative-HPLC (2 citations)

149 protocols using 1260 hplc

Quantitative Analysis of ADC Characteristics

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HIC HPLC was used to determine levels of the molar ratio of drug substitution: 1260 HPLC (Agilent; Wilmington, DE, USA); Butyl-NPR column (2.5 μm, 4.6 × 35 mm, #14947, TOSOH Bioscience; Tokyo, Japan). Here, HIC buffer A was 50 mM potassium phosphate, pH 7.0, and 1.5 M ammonium sulfate; and HIC buffer B was 50 mM potassium phosphate, pH 7.0, 20% isopropanol. The gradient was 100% to 100% B over 15 min; flow rate was 1 mL/min; and UV detection wavelength was 280 nm. The DAR was determined by peak area integration according to the reported method [32 ].
SEC HPLC was used to determine levels of aggregation within each ADC: 1260 HPLC (Agilent; Wilmington, DE, USA); G3000SWXL analytical column (7.8 mm × 30 cm, #08541, TOSOH Bioscience; Tokyo, Japan). The SEC buffer contained 40 mM sodium phosphate and 150 mM sodium chloride (pH 7.0, 1 mL/min flow rate); and the UV detection wavelength was 280 nm. We performed a needle wash after each injection and include blank runs between each analyte. The aggregation was determined by peak area integration according to the reported method [33 ].

Wang Y., Fan S., Zhong W., Zhou X, & Li S. (2017). Development and Properties of Valine-Alanine based Antibody-Drug Conjugates with Monomethyl Auristatin E as the Potent Payload. International Journal of Molecular Sciences, 18(9), 1860.

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SEC-UV and SEC-MALS Protein Analysis

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SEC-UV method consisted of an Agilent 1260 HPLC with a UV-Vis detector. An XBridge Protein BEH SEC Analytical Column (Waters, 450 Å, 3.5 µm, 7.8 × 300 mm) was used for the separation. An XBridge Protein BEH SEC Guard Column (Waters, 450 Å, 3.5 µm, 7.8 × 30 mm) was attached before the analytical column as a protection. The column was kept at 30°C. The UV detection was set at 220 nm. The flow rate was 1 mL/min. The SEC-MALS method consisted of an Agilent 1260 HPLC with UV, DAWN (Wyatt) MALS, and Optilab (Wyatt) relative refractive interferometer (RI). A Tosoh TSKgel G4000SWXL column (P/N 08542, 8 µm, 7.8 × 300 mm) was used for the separation. The flow rate was 0.4 mL/min. The column and mobile phase were held at room temperature. Data were analysed using ASTRA software. Peak alignment and band broadening correction between UV, MALS, and RI detectors were performed. The dn/dc of 0.185 was used for the protein fraction and 0.147 for the glycan modifier.

Guo H., Cho B., Hinton P.R., He S., Yu Y., Ramesh A.K., Sivaccumar J.P., Ku Z., Campo K., Holland S., Sachdeva S., Mensch C., Dawod M., Whitaker A., Eisenhauer P., Falcone A., Honce R., Botten J.W., Carroll S.F., Keyt B.A., Womack A.W., Strohl W.R., Xu K., Zhang N., An Z., Ha S., Shiver J.W, & Fu T.M. (2023). An ACE2 decamer viral trap as a durable intervention solution for current and future SARS-CoV. Emerging Microbes & Infections, 12(2), 2275598.

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HPLC Quantitation of Phenylalanine

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An aliquot of an ongoing aggregation reaction was subjected to centrifugation at 305,611×g for 2 h. 30 μL supernatant was subjected for analysis at 215 nm wavelength on Agilent HPLC 1260 using ZORBAX Eclipse Plus C18 Rapid Resolution (4.6 × 100 mm, 3.5 μm) column with water (mobile phase A) and acetonitrile (mobile phase B) containing 0.05% TFA at flow rate of 1 mL min⁻¹ for 20 minutes. Mobile phase B was increased from 0–30% in 10 minutes and Phe eluted at 6.0 ± 0.3 minute between 8.8–10.25% of mobile phase B.

Singh V., Rai R.K., Arora A., Sinha N, & Thakur A.K. (2014). Therapeutic implication of L-phenylalanine aggregation mechanism and its modulation by D-phenylalanine in phenylketonuria. Scientific Reports, 4, 3875.

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HPLC Analysis of Carotenoids in Samples

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Extracts were analyzed by HPLC with an automatic injection pump (Agilent Technology HPLC 1260) and diode array detector (model G4212B) in 5 μm columns (ZORBAX SB-C8, 4.6 cm x150 mm). The mobile phase was activated by a quaternary pump (model VL 1260). Solvents used were: solvent A, methanol, 1 N ammonium acetate (70:30 v/v); and solvent B, methanol at a flow rate of 1 mL min⁻¹ for 20 min at room temperature. The gradient system was according the following procedure (minutes; %solvent A, % solvent B): (0; 75, 25), (1; 50, 50), (15; 0, 100), (18; 0, 100), and (18.5; 75, 25) [57 ]. astaxanthin and other carotenoids were identified at 440 nm and compared against known standards for retention time and absorption spectrum. Known standards were astaxanthin, zeaxanthin, β-carotene, canthaxanthin, and chlorophyll-a (Sigma-Aldrich).

Galarza J.I., Arredondo Vega B.O., Villón J, & Henríquez V. (2019). Deesterification of astaxanthin and intermediate esters from Haematococcus pluvialis subjected to stress. Biotechnology Reports, 23, e00351.

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HPLC Analysis of Organic Compounds

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HPLC analysis were performed on an Agilent HPLC 1260 apparatus, equipped with ZORBAX Eclipse XDB-C8 reverse phase column. The mobile phase consisted in a 1:1, v/v mixture of acetonitrile and water, with a constant flow rate of 0.5 mL/min. HPLC solvents were purchased from Carlo Erba reagents (HPLC quality) and water was buffered with trifluoroacetic acid (0.5%, v). HPLC chromatograms were obtained with UV detection at 254 nm.

Rouillon J., Arnoux C, & Monnereau C. (2021). Determination of Photoinduced Radical Generation Quantum Efficiencies by Combining Chemical Actinometry and ¹⁹F NMR Spectroscopy. Analytical chemistry, 93(5).

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Phenolic Compounds Analysis via HPLC-ESI-QTOF-MS/MS

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Phenolic compounds analyses in PE, microparticles, and digested samples were carried out using High-Performance Liquid Chromatography Coupled to Electrospray Quadrupole-Time-of-Flight Mass Spectrometry (HPLC-ESI-QTOF-MS/MS) (HPLC 1260 coupled to 6540 Ultra High Definition Accurate-Mass Q-TOF, Agilent Technologies, Palo Alto, CA, USA) with the method described elsewhere [25 (link)]. Briefly, the separation was performed with a Agilent Zorbax Eclipse Plus C18 column (150 mm × 4.6 mm, 1.8 µm) using as mobile phases water with 0.1% formic acid and acetonitrile in gradient elution mode. Detection was performed in negative ion mode within a mass range of 50–1700 m/z.

Cea-Pavez I., Manteca-Bautista D., Morillo-Gomar A., Quirantes-Piné R, & Quiles J.L. (2024). Influence of the Encapsulating Agent on the Bioaccessibility of Phenolic Compounds from Microencapsulated Propolis Extract during In Vitro Gastrointestinal Digestion. Foods, 13(3), 425.

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Enzymatic Conversion of Hydroxybenzoic Acids

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Conversion of (hydroxy)benzoic acids was additionally analyzed by LC–MS. Assays were performed using 100 µl reaction volume consisting of 100 mM K₂HPO₄/KH₂PO₄, pH 7.0, 7.5 µg (Aa4CL) or 28 µg (Aa4HBCL) purified enzyme, 1.25 mM ATP, 1.25 mM MgCl₂, 500 µM (hydroxy)benzoic acid derivative and 1 mM CoA. Assays were incubated at 40 °C for 30 min (Aa4HBCL, Aa4CL with 2-coumaric and 3-coumaric acid) or 60 min at 35 °C (Aa4CL with hydroxybenzoic acids). An assay with heat-denatured protein (10 min at 95 °C) served as negative control. Assays were stopped on ice by the addition of 100 µl methanol. After centrifugation for 10 min at 15,000g, 15 µl of the supernatant was analyzed by LC–MS. LC was performed on a HPLC 1260 (Agilent Technologies) with a Multospher 120 RP18 column (250 × 2 mm; particle size 5 μm) using a solvent system of A = 0.1% (v/v) aqueous formic acid, B = acetonitrile with 0.1% (v/v) formic acid at a flow rate of 0.5 ml/min and a temperature of 25 °C with the following gradient: 0–10 min 5% B → 100% B; 10–15 min 100% B; 15–15.1 min 100% B → 5% B; 15.1–20 min 5% B. Detection was performed with the mass spectrometer micrOTOF-Q III with ESI source (Bruker Daltonics) calibrated with 5 mM sodium formate using the negative mode.

Wohl J, & Petersen M. (2020). Phenolic metabolism in the hornwort Anthoceros agrestis: 4-coumarate CoA ligase and 4-hydroxybenzoate CoA ligase. Plant Cell Reports, 39(9), 1129-1141.

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Assessing p-NP Degradation Kinetics

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The concentration of p-NP was determined using high-performance liquid chromatography (HPLC 1260, Agilent, US) obeying method depicted in ESI.† To clarify the effects of environmental parameters on p-NP degradation kinetics in the heterogeneous Fenton reaction, the experimental data obtained in the early stage were fitted using pseudo-first-order kinetics (eqn (S1)†), and their rate constants were calculated and compared. The concentrations of ferrous ion and total iron were measured with 1,10-phenanthroline following standard method at 510 nm using a UV/Vis spectrophotometer (UV-1800, Shimadzu, Japan). The electron spin resonance (ESR) experiment was conducted using MS-5000 spectrometer (Magnettech, Germany) and DMPO as ˙OH trapper with sweep width, microwave frequency, power set at 332.5–342.5 mT, 9.46 GHz, and 10 mW, respectively. The degradation intermediates of p-NP were analyzed using GC-MS (GC-2014, Shimadzu, Japan).

Liu T., Chen N., Deng Y., Chen F, & Feng C. (2020). Degradation of p-nitrophenol by nano-pyrite catalyzed Fenton reaction with enhanced peroxide utilization. RSC Advances, 10(27), 15901-15912.

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Chiral Benzonitrile Derivative Synthesis

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2-Bromo-4-[2,6-dioxo-4-(trifluoromethyl)-3,6-dihydropyrimidin-1(2H)-yl]-5-methoxybenzonitrile (91, 600 mg, 1.54
mmol), 2-methylphenol (220 μL, 2.20 mmol), N,N-dimethylglycine (42.0 mg, 408 μmol), copper(I)
iodide (39.5 mg, 208 μmol), and cesium carbonate (1.20 g, 3.69
mmol) were heated in DMF (30 mL) for 18 h at 140 °C. Upon completion
of the reaction, work-up, and purification, the title compound was
obtained (35 mg, 6%) as an ochre solid. ¹H NMR (400 MHz,
[D]₆ DMSO): δ 2.18 (s, 3 H), 3.79 (s, 3H), 6.26 (br
s, 1 H), 6.97 (m, 2H), 7.17, 7.27, 7.35 (3 m, 1H each), 7.73 (s, 1H),
12.56 (br s, 1H); LCMS (ESI−) m/z: 416 [M – H]⁻. Atropisomeric ratio: atrop
1/atrop 2 = 50:34 (16% impurities); ^tR (atrop1) = 3.20 min; ^tR (atrop2) = 4.99 min. The atropisomeric ratio was determined
using the following chiral HPLC method: instrument: Agilent HPLC 1260;
column: Chiralpak ID 3 μm 100 × 4.6 mm; eluent A: hexane
+ 0.1 vol % diethylamine (99%); eluent B: 2-propanol; gradient: 20–50%
B in 7 min; flow: 1.4 mL/min; temperature: 25 °C; DAD 254 nm.

Günther J., Hillig R.C., Zimmermann K., Kaulfuss S., Lemos C., Nguyen D., Rehwinkel H., Habgood M., Lechner C., Neuhaus R., Ganzer U., Drewes M., Chai J, & Bouché L. (2022). BAY-069, a Novel (Trifluoromethyl)pyrimidinedione-Based BCAT1/2 Inhibitor and Chemical Probe. Journal of Medicinal Chemistry, 65(21), 14366-14390.

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Atropisomeric Synthesis of Substituted Naphthalene

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Aryl bromide 12 (100 mg, 0.222
mmol), piperidine (29 μL, 0.244 mmol), and potassium carbonate
(77 mg, 0.555 mmol) in DMSO (4 mL) gave 24 as a yellow
solid (25 mg, 24%). ¹H NMR (400 MHz, [D]₆DMSO):
δ = 1.68, 1.85 (2 m_c, 2 H, 4 H), 2.06 (s, 3 H), 3.24
(m_c, 4 H), 3.28, 3.37 (2 s, 3 H), 7.13, 7.16 (2 m_c, 1 H each), 7.36 (2 s, 1 H each), 7.85 (dd, J =
7.4, 8.4 Hz, 1 H), 8.41 (d, J = 8.1 Hz, 1 H), 8.49
(m_c, 2 H each) ppm; LCMS (method III): ^tR = 1.42 min; m/z: [M + H]⁺ = 455; atropisomeric ratio: atrop1/atrop2 = 50:50; ^tR (atrop1) = 3.60 min; ^tR (atrop2) = 4.59 min. The atropisomeric ratio was determined using
the following chiral HPLC method: instrument: Agilent HPLC 1260; column:
Chiralpak ID 3 μm 100 × 4.6 mm; eluent A: 2-methoxy-2-methylpropane
+ 0.1 vol % diethylamine (99%); eluent B: ACN; isocratic: 50% A +
50% B; flow: 1.4 mL/min; temperature: 25 °C; DAD: 254 nm.

Bouché L., Christ C.D., Siegel S., Fernández-Montalván A.E., Holton S.J., Fedorov O., ter Laak A., Sugawara T., Stöckigt D., Tallant C., Bennett J., Monteiro O., Díaz-Sáez L., Siejka P., Meier J., Pütter V., Weiske J., Müller S., Huber K.V., Hartung I.V, & Haendler B. (2017). Benzoisoquinolinediones as Potent and Selective Inhibitors of BRPF2 and TAF1/TAF1L Bromodomains. Journal of Medicinal Chemistry, 60(9), 4002-4022.

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