Alliance e2695

Manufactured by Waters Corporation

Sourced in United States, Japan

The Alliance e2695 is a high-performance liquid chromatography (HPLC) system manufactured by Waters Corporation. It is designed to deliver consistent and reliable separation of complex samples. The Alliance e2695 features advanced solvent and sample management systems to provide precise control and reproducible results.

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51 protocols using alliance e2695

Extraction and Analysis of Placental and Pericarp Phytochemicals

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The placentas were collected at 45 DPA and were oven-dried at 75 for 48 h. Fine powder (0.1 g) of the ovendried placenta was added into 5 ml of methyl alcohol and tetrahydrofuran (1:1, HPLC grade) and then was extracted for 12h at room temperature. The supernatant was filtered using a 0.22 μm Millipore membrane. The capsaicinoids content was measured by a high-performance liquid chromatography (HPLC) system (Alliance E2695, Waters, America).
The pericarps were collected at 45 DPA and freeze-dried for 24 h in a freeze dryer (Labconco/FreeZone, Labconco, America). freeze-dried pericarp (0.5 g) was added into 2 ml of absolute ethyl alcohol, 2 ml of acetone and 4 ml of hexyl hydride (HPLC grade). Five millilitres of supernatants were added to 10 ml NaCl saturated solution (100%), 0.2 ml KOH and 1.8 ml methyl alcohol and then incubated at room temperature for 12 h. The extract was added to 2 ml of MTBE and NaCl saturated solution (100%). NaCl saturated solution (100%) was used for rinsing the supernatants three times. The supernatants were filtered using a 0.22 μm Millipore membrane. The determination of carotenoid content was performed by an HPLC system (Alliance E2695, Waters, America).

Liu R., Song J., Liu S., Chen C., Zhang S., Wang J., Cao B., Zhu Z., & Lei J. (2020). Genome-wide identification of the Capsicum bHLH transcription factor family: discovery of candidate regulator involved in the regulation of species-specific bioactive metabolites.

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Quantifying Lignin Monomers by Alkaline Nitrobenzene Oxidation

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The alkaline nitrobenzene oxidation (NBO) method was used to analyze the monolignin proportion as described by Wu et al. [53 (link)]. HPLC (Waters e2695 Alliance) with a column of Kromat Universil C₁₈ (4.6 mm × 250 mm, 5 μm) was used to analyze the lignin monomers using p-Hydroxybenzaldehyde (H), Vanillin (G) and Syringaldehyde (S) (purchased from Sinopharm Chemical Reagent Co., Ltd.) as standard chemicals and ethyl vanillin as the internal standard material. The column was operated at 30°C with a CH₃OH : H₂O : CH₃COOH (23 : 76 : 2, v / v / v) carrier liquid at a flow rate of 1.0 mL/min. The calibration curves of all analytes routinely yielded correlation coefficients 0.999 or higher.

Zhao C., Fan X., Hou X., Zhu Y., Yue Y., Zhang S, & Wu J. (2015). Tassel Removal Positively Affects Biomass Production Coupled with Significantly Increasing Stem Digestibility in Switchgrass. PLoS ONE, 10(4), e0120845.

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HPLC Analysis of A7OG and Baicalin

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The HPLC system with a Shimadzu SPD-M20A photodiode array detector (Shimadzu, Kyoto, Japan) was used for the analysis and validation. The Waters e2695 alliance (Waters Corporation, Milford, MA, USA) HPLC system was used for the inter-laboratory validation. The temperature of the column (YoungJin Biochrom INNO-P C18 5 µm, 4.6 × 150 mm) was set at 30 °C, and the flow rate was maintained at 1 mL/min. The injection volume was 10 µL, and the detection wavelength was 335 nm. The mobile phase consisted of (A) distilled water with 0.1% phosphoric acid and (B) acetonitrile. For the separation of A7OG and baicalin, the following gradient was used: solution B was changed from 90% to 75% between 0 and 30 min; then, B was 75% to 90%, from 30 to 40 min, and isocratic from 40 to 50 min.

Lee J.S., Nam Y.R., Kim H.J, & Kim W.K. (2023). Quantification and Validation of an HPLC Method for Low Concentrations of Apigenin-7-O-Glucuronide in Agrimonia pilosa Aqueous Ethanol Extract Topical Cream by Liquid–Liquid Extraction. Molecules, 28(2), 713.

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Ethanol content analysis in beer

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The ethanol content in beer was analysed with a Waters e2695 Alliance liquid chromatograph (Waters, Massachusetts, USA) equipped with a refractive index detector 2414 and a photodiode array detector 996. MilliQ water was used as the solvent with a flow rate of 0.4 mL/min in isocratic mode. 200 µm of each beer sample was removed for pre-filtering through 0.45 µm pore size cellulose ester filters (Teknokroma, Barcelona, Spain) and then a total of 25 samples were injected into the HPLC apparatus. Separation of the analytes was carried out on a C18 PhenoSphere XDB reverse phase column (150 × 4.6 mm, 5 µm particle size) (Phenomenex, California, USA) stabilized at 30 °C. Quantification was performed with ethanol (99.5% purity) (Panreac, Spain) as an external standard with four calibration levels: 5, 10, 15 and 20% v/v (r 2 = 0.9734). The injection volume was 2 µl. The data were collected in a table and the mean value of each species was calculated for comparison.

Callejo M.J., García Navas J.J., Alba R., Escott C., Loira I., González M.C, & Morata A. (2019). Wort fermentation and beer conditioning with selected non-Saccharomyces yeasts in craft beers. European food research and technology = Zeitschrift fur Lebensmittel-Untersuchung und -Forschung. A, 245(6).

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Molecular Weight Analysis of TGA Solution

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TGA solution was
added to make a 10 mg/mL solution under the condition of a highly
pure water mobile phase and a flow rate of 0.8 mL/min. The solution
was first ultrasonicated for 30 min at room temperature and then allowed
to stand for 3 h to make it fully dispersed. After shaking well, the
solution was centrifuged at 5000 r/min for 15 min, and the supernatant
was taken to measure its molecular weight by gel chromatography (Alliance
e2695, Waters, USA).

Tian Y., Liu X., Luo P., Huang J., Xiong J., Liang L, & Li W. (2021). Study of a Polyamine Inhibitor Used for Shale Water-Based Drilling Fluid. ACS Omega, 6(23), 15448-15459.

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Quantification of Residual Drugs and Metabolites

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The concentration of residual RBV in the filtrate was determined using a high-performance liquid chromatography system (Alliance e2695, Waters, USA), equipped with a UV–vis detector and a C18 column (4.6 mm × 250 mm, 5 μm) at 30 °C. The mobile phase consisted of 10 mM potassium dihydrogen phosphate (pH ≈ 5.1) at a flow rate of 1 mL·min⁻¹. The injection volume was 10 μL, and the detection wavelength was 207 nm.
The TPs were detected using a UHPLC-Q-Orbitrap HRMS system (Dionex Ultimate 3000 Series; MS, Thermo Q Exactive Plus). Chromatographic separation was conducted using an ACQUITY UPLC HSS T3 C18 column (2.1 mm × 100 mm, 1.8 μm; Waters, USA) maintained at 30 °C. Gradient elution was performed with 0.1 % (v/v) formic acid in water (solvent B) and acetonitrile (solvent A) at a flow rate of 0.2 mL·min⁻¹. Further details regarding the analytical methods are provided in the Supplementary material (Text S1).

Wu X., Zhang J., Hu S., Zhang G., Lan H., Peng J, & Liu H. (2022). Evaluation of degradation performance toward antiviral drug ribavirin using advanced oxidation process and its relations to ecotoxicity evolution. The Science of the Total Environment, 850, 157851.

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HPLC Characterization of SC Extract

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We used a Waters HPLC system with a Waters alliance e2695 and a Waters 2998 photodiode array detector. The Fortis C18 analytical column (150 × 4.6 mm, 5 µm pore size) was tested and filled with the same stationary phase. Samples A (0.01% Formic acid) and B (ACN) were used as the mobile phase under gradient conditions to analyze the samples. The mobile phase was filtered by vacuum through a 0.45 µm membrane filter. Chromatography was conducted on a gradient using a flow rate of 1.0 ml/min at 30 °C and was detected at various UV wavelengths for individual standards and SC samples. The SC extract powder was dissolved in 50% MeOH at 10 mg/ml. The chromatograms were processed using Empower 2 software, build 1154 (Waters, Milford, MA, USA).

Song M., Lee S.Y., Kim M., Park S., Park J., Kwon Y, & Park D.H. (2020). Saururus chinensis-controlled allergic pulmonary disease through NF-κB/COX-2 and PGE2 pathways. PeerJ, 8, e10043.

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Polymer Molecular Weight Analysis

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GPC measurements in Figure 2 were obtained with
a GPC Waters Alliance e2695 instrument equipped with a refractive-index
RI 2414 detector and Waters Ultrahydrogel 120, 250, 500 in series
columns. H₂O/NaNO₃/NaOH (0.1 M) was used as
an eluent at a flow rate of 0.7 mL/min at 40 °C. Data processing
was performed with Empower 3 software. The molecular weight of the
polymers was calculated on the basis of PEG calibration curve.

Capacchione C., Della Sala P., Quaratesi I., Bruno I., Pauciulo A., Bartiromo A.R., Iannece P., Neri P., Talotta C., Gliubizzi R, & Gaeta C. (2021). Poly(Ethylene Glycol)/β-Cyclodextrin Pseudorotaxane Complexes as Sustainable Dispersing and Retarding Materials in a Cement-Based Mortar. ACS Omega, 6(18), 12250-12260.

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HPLC-SEC Characterization of Polymers

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The HPLC-SEC system included an Alliance e2695 module, Refractive Index (RI) 2414 detector, Empower 3 software and Ultrahydrogel 2000 column (7.8 × 300 mm) Waters (Milford, MA, USA), 0.1 M sodium nitrate mobile phase at a flow rate of 0.8 mL/min and 80 µL was injected for analysis. The method used was adapted from Jagannath and Ramachandran [33 (link)].

Suárez-Hernández L.A., Camacho-Ruíz R.M., Arriola-Guevara E., Padilla-Camberos E., Kirchmayr M.R., Corona-González R.I, & Guatemala-Morales G.M. (2021). Validation of an Analytical Method for the Simultaneous Determination of Hyaluronic Acid Concentration and Molecular Weight by Size-Exclusion Chromatography. Molecules, 26(17), 5360.

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Quantification of Plasma BH4 by HPLC

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BH4 levels were determined by high performance liquid chromatography (HPLC) and quantified using electrochemical detection as previously described with some modifications (Latini et al., 2018 (link)). Plasma samples were thawed on ice and protected from light. Afterwards, samples were centrifuged (16,000 × g; 10 min; 4 °C) and 20 μL of supernatant were transferred to an HPLC vial for analysis. The HPLC analysis of BH4 was carried out in a HPLC system (Alliance e2695, Waters, Milford, USA) by using a Waters Atlantis dC18 reverse phase column (4.6 × 250 mm; 5 μm particle), with the flow rate set at 0.7 mL/min and an isocratic elution of 6.5 mM NaH₂PO₄, 6 mM citric acid, 1 mM sodium octyl sulfate, 2.5 mM diethylenetriaminepentaacetic acid, 160 μM dithiothreitol and 8 % acetonitrile, at pH 3.0. The temperature of the column compartment was set at 35 °C. The identification and quantification of BH4 was performed by an electrochemical detector (module 2465, Waters, Milford, USA) using a voltage of +450 mV. The results were expressed as ηmol/L.

Cronin S.J., Yu W., Hale A., Licht-Mayer S., Crabtree M.J., Korecka J.A., Tretiakov E.O., Sealey-Cardona M., Somlyay M., Onji M., An M., Fox J.D., Turnes B.L., Gomez-Diaz C., da Luz Scheffer D., Cikes D., Nagy V., Weidinger A., Wolf A., Reither H., Chabloz A., Kavirayani A., Rao S., Andrews N., Latremoliere A., Costigan M., Douglas G., Freitas F.C., Pifl C., Walz R., Konrat R., Mahad D.J., Koslov A.V., Latini A., Isacson O., Harkany T., Hallett P.J., Bagby S., Woolf C.J., Channon K.M., Je H.S, & Penninger J.M. (2023). Crucial neuroprotective roles of the metabolite BH4 in dopaminergic neurons. bioRxiv.

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