1525 binary hplc pump

Manufactured by Waters Corporation

Sourced in United States, United Kingdom, Japan, Ireland

The 1525 binary HPLC pump is a high-performance liquid chromatography (HPLC) pump designed to deliver precise and consistent solvent flow rates. It features a binary solvent delivery system that can handle two different solvents simultaneously. The pump is capable of generating a wide range of flow rates and pressures, making it suitable for a variety of HPLC applications.

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Lab products found in correlation

143 protocols using 1525 binary hplc pump

Peptoid Synthesis via Microwave-Assisted SPPS

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All the Fmoc-protected amino acids were purchased from AnaSpec
(Fremont, CA). TentaGel resin was purchased from Rapp Polymere GmbH
(German). All other reagents were purchased from Sigma-Aldrich or
Alfa Aesar, unless otherwise specified. All of the chemical reagents
and solvents from commercial sources were used without further purification.
Five mL and 10 mL disposable reaction columns (Intavis AG) were used
as reaction vessels for solid-phase synthesis. Syntheses of peptiods
under microwave conditions were performed in a 1500 W microwave oven
(GE model JE 1860BH04) with 10% power. HPLC was carried out on Waters
systems equipped with Waters 1525 binary HPLC pumps and a 2487 dual
λ
absorbance detector, or a 2998 photodiode array detector. The mobile
phase comprised of buffer A (H₂O containing 20% CH₃CN and 0.1% trifluoroacetic acid (TFA)) and buffer B (CH₃CN containing 0.1% TFA). Analytical HPLC was conducted using
a Vydac C-18 column (5 μm, 250 × 4.6 mm, Alltech, Deerfield,
IL) at a flow rate of 1.0 mL/min with UV detection at 220 nm. MS and
MS/MS (MALDI-TOF) were performed on a 4800 Proteomics Analyzer (Applied
Biosystems) with α-cyano-4-hydroxycinnamic acid (CHCA) as a
matrix. All steps involving water utilized distilled water filtered
through a Barnstead Nanopure filtration system (Thermo Scientific)

Gao Y., Amar S., Pahwa S., Fields G, & Kodadek T. (2014). Rapid Lead Discovery Through Iterative Screening of One Bead One Compound Libraries. ACS Combinatorial Science, 17(1), 49-59.

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HPLC Analysis of Organic Compounds

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Chromatographic analysis was performed on a Waters liquid chromatography system, equipped with a Waters 2996 photodiode array detector, Waters 1525 binary HPLC pumps, a Waters in-line degasser AF and a Waters 717 plus autosampler. The system was controlled with Empower software. Chromatographic separations were performed using a Purospher^®Star RP-18 end-capped C18 column (4.6 × 150 mm, 5 μm pore size, Merck KGaA, Damstadt, Germany) at ambient temperature. The monitoring wavelength was set to 271 nm. An elution gradient over 60 mL from 0 to 100% buffer B (Buffer A: 20 mM sodium dihydrogen phosphate/5 mm phosphoric acid pH 2.6 in water; buffer B: 100% acetonitrile) was used at a flow rate of 1 mL min⁻¹. 20-μL samples were injected using a sample loop. At the end of each run, the column was rinsed by a cleanup procedure with pure acetonitrile.

Krueger A., Baumann S., Krammer P.H, & Kirchhoff S. (2001). FLICE-Inhibitory Proteins: Regulators of Death Receptor-Mediated Apoptosis. Molecular and Cellular Biology, 21(24), 8247-8254.

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Quantitative Analysis of Mebendazole in Biological Samples

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For MBZ assay, 0.5 ml plasma, cyst wall, cyst fluid and mice’s tissue samples with ABZ as internal standard was extracted with Oasis HLB Cartridges (Waters, USA). The final collection of methanol elution was dried and redissolved in methanol. All samples were filtrated with 0.45 μm membrane filter before injection.
The system of instrument consisted of a 1525 Binary HPLC Pumps, a 717 plus auto sampler and a 2457 Dual λ Absorbance Detector (Waters, USA). The assay of MBZ was performed in a 5 μm C₁₈ 250 × 4.6 mm column (Beckman Coulter, USA) and a mobile phase containing 350 ml of 0.05 M ammonium sulfate and 650 ml of methanol at a flow rate of 1 ml/min. The MBZ concentration was measured by its absorbance at a wavelength of 289 nm. The details and validation of the analytical methodology can be seen in Additional file 1.

Liu C.S., Zhang H.B., Lei W., Zhang C.W., Jiang B., Zheng Q., Yin J.H, & Han X.M. (2014). An alternative mebendazole formulation for cystic echinococcosis: the treatment efficacy, pharmacokinetics and safety in mice. Parasites & Vectors, 7, 589.

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HPLC Analysis of Pharmaceutical Compounds

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Chromatographic analysis was performed on a Waters liquid chromatography system, equipped with a Waters 2996 photodiode array detector, Waters 1525 binary HPLC pumps, a Waters in-line degasser AF and a Waters 717 plus autosampler. The system was controlled with Empower software. Chromatographic separations were performed using a Purospher ® Star RP-18 end-capped C18 column (4.6 × 150 mm, 5 μm pore size, Merck KGaA, Damstadt, Germany) at ambient temperature. The monitoring wavelength was set to 271 nm. An elution gradient over 60 mL from 0 to 100% buffer B (Buffer A: 20 mM sodium dihydrogen phosphate/5 mm phosphoric acid pH 2.6 in water; buffer B: 100% acetonitrile) was used at a flow rate of 1 mL min -1 . 20-μL samples were injected using a sample loop. At the end of each run, the column was rinsed by a cleanup procedure with pure acetonitrile.

Khongthong S., Theapparat Y., Roekngam N., Tantisuwanno C., Otto M, & Piewngam P. (2021). Characterization and immunomodulatory activity of sulfated galactan from the red seaweed Gracilaria fisheri. International journal of biological macromolecules, 189.

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Spectroscopic Analysis of Chemical Compounds

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Optical rotations were measured using a Jasco P-1010 Polarimeter with sodium light (589 nm). UV spectra were recorded with a Waters 2998 Photodiode Array Detector. ¹H and ¹³C NMR spectra were recorded on a Bruker AV-600 spectrometer with a 5 mm CPTCI cryoprobe. ¹H chemical shifts are referenced to the residual DMSO-d₆ (δ 2.49 ppm) and ¹³C chemical shifts are referenced to the DMSO-d₆ solvent peak (δ 39.5 ppm). Low and high-resolution ESI-QIT-MS were recorded on a Bruker-Hewlett Packard 1100 Esquire–LC system mass spectrometer. Merck Type 5554 silica gel plates and Whatman MKC18F plates were used for analytical thin-layer chromatography. Reversed-phase HPLC purifications were performed on a Waters 1525 Binary HPLC Pump attached to a Waters 2998 Photodiode Array Detector. All solvents used for HPLC were Fisher HPLC grade.

Williams D.E., Morgan K.D., Dalisay D.S., Matainaho T., Perrachon E., Viller N., Delcroix M., Gauchot J., Niikura H., Patrick B.O., Ryan K.S, & Andersen R.J. (2022). Natural Products Produced in Culture by Biosynthetically Talented Salinispora arenicola Strains Isolated from Northeastern and South Pacific Marine Sediments. Molecules, 27(11), 3569.

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HPLC Separation of Theobromine and Metabolites

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Chromatographic separation was done at 27 °C on an Atlantis dC18 5 µm column (part no. 186001344) fitted to a Waters HPLC unit consisting of a Waters In-Line degasser AF, Waters 1525 binary HPLC pump and Waters 2487 dual wavelength absorbance detector set to monitor at 270 nm. Gradient elution at 1.5 ml/min was by a solvent system of acetonitrile (Sigma-Aldrich; St. Louis, MO) and 0.1% formic acid (BDH, England). The formic acid solution was adjusted to pH 3.75 with ammonia (BDH; Poole, England). Mobile phases were pumped according to the gradient profile in Table 1. Solutions of theobromine, as well as theophylline and caffeine (Sigma-Aldrich; St. Louis, MO) were prepared and used as standards.Table 1

Gradient elution profile for separation of theobromine and its metabolites

Time (min)	Eluent mix (%)		Flow rate (ml/min)
Time (min)	0.1% ammonium formate	acetonitrile	Flow rate (ml/min)
0	98	2	1.5
6	95	5	1.5
9	98	2	1.5
10	98	2	1.5
11	92	8	1.5
14	88	12	1.5
17	89	11	1.5
18	98	2	1.5

Oduro-Mensah D., Ocloo A., Lowor S.T., Mingle C., Okine L.K, & Adamafio N.A. (2018). Bio-detheobromination of cocoa pod husks: reduction of ochratoxin A content without change in nutrient profile. Microbial Cell Factories, 17, 79.

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Peptide Synthesis and Purification Protocol

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Peptides (GRGDSPCG, GCALMKYH ILNTLQCSE, GCDPGIVRRADRAAVP, GCDPGIKVAV, GCDPGYISGR, GCGDGEA, GCGFYFDLR, CSVTCG, CGGAEIDGIEL, GCRDIPVSLRSGDRCG, GCRDRPFSMIMGDRCG, GCRDVPLSLTMGDRCG, GCRDVPLSLYSGDRCG, and GCRDIPESLRAGDRCG) were synthesized on a CEM Liberty Blue automated solid phase peptide synthesizer (CEM, Mathews, NC) using FMOC protected amino acids (Iris Biotech GMBH, Germany). The peptide was cleaved from the resin by sparging-nitrogen gas through a solution of trifluoroacetic acid (TFA), triisopropylsilane (TIPS), 2,2′(ethylenedioxy)diethanethiol (DODT), and water at a ratio of 92.5:2.5:2.5:2.5 vol%, respectively (Sigma-Aldrich) for 2–3 h at RT in a reactor vessel (ChemGlass, Vineland, NJ). After reaction, the solution was filtered, and the peptide was precipitated using ethyl ether at −80 °C (Thermo). The molecular weight of the peptide was validated using a MicroFlex MALDI-TOF (Bruker, Billerica, MA) using alpha-cyano-4-hydroxycinnamic acid as the matrix (Sigma-Aldrich). Peptides were analyzed and purified to >95% on a VYDAC reversed-phase C18 column attached to a Waters 2487 dual (lambda) adsorbable detector and 1525 binary HPLC pump (Waters, Milford, MA).

Galarza S., Crosby A.J., Pak C, & Peyton S.R. (2020). Control of Astrocyte Quiescence and Activation in a Synthetic Brain Hydrogel. Advanced healthcare materials, 9(4), e1901419.

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Synthesis and Characterization of Bleomycin-Derived Compounds

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Bleomycin A5 (Bleocin) was purchased from EMD Millipore and used without further purification. Hoechst carboxylic acid, 4-(3-(6-(4-methylpiperazin-1-yl)-1H,3′H-[2,5′-bibenzo[d]imidazol]-2′-yl)phenoxy)butanoic acid (6) (Ht-CO₂H), intermediate (10) and 2H-K4NMeS were synthesized as reported previously.^{14 (link)} Peptide synthesis reactions were monitored by a chloranil test. Preparative HPLC was performed using a Waters 1525 Binary HPLC pump equipped with a Waters 2487 dual absorbance detector system and a Waters Sunfire C18 OBD 5 μm 19 × 150 mm column. Absorbance was monitored at 345 and 220 nm or 254 nm. A gradient of methanol in water with 0.1% TFA varied in each purification. Purity was assessed by analytical HPLC using a Waters Symmetry C18 5 μm 4.6 × 150 mm column, and a linear gradient of 0–100% methanol in water with 0.1% TFA over 60 min. Absorbance was monitored at 345 and 254 nm or 220 nm. All compounds evaluated had ≥95% purity (see Characterization of Small Molecules). Mass spectrometry was performed with an Applied Biosystems MALDI ToF/ToF Analyzer 4800 Plus using an α-cyano-4-hydroxycinnamic acid matrix or an Agilent 1260 Infinity LC system coupled to an Agilent 6230 TOF (HR-ESI) with a Poroshell 120 EC-C18 column (Agilent, 50 mm × 4.6 mm, 2.7 μm).

Benhamou R.I., Abe M., Choudhary S., Meyer S.M., Angelbello A.J, & Disney M.D. (2020). Optimization of the linker domain in a dimeric compound that degrades an r(CUG) repeat expansion in cells. Journal of medicinal chemistry, 63(14), 7827-7839.

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Spectroscopic Analysis of Chemical Compounds

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Melting points were determined with a Mel-Temp apparatus using capillary tubes and are uncorrected. Proton nuclear magnetic resonance spectra (¹H NMR) were recorded using an ARX300 300 MHz Bruker NMR spectrometer. IR spectra were obtained with a PerkinElmer 1600 series FTIR spectrometer. The purities of all of the biologically tested compounds were estimated by HPLC, and in each case, the major peak accounted for ≥95% of the combined total peak area when monitored by a UV detector at 254 nm. HPLC analyses were performed on a Waters 1525 binary HPLC pump/Waters 2487 dual λ absorbance detector system. HPLC analyses were performed on a Sunrise C-18 column with dimensions of 16 × 4.6 cm and 5 μm particle size. Analytical thin-layer chromatography was conducted on Baker-flex silica gel IB2-F plates, and compounds were visualized with UV light at 254 nm. Silica gel flash chromatography was performed using 230–400 mesh silica gel. The anhydrides 11a and 11b and compound 12 were prepared according to the literature and showed similar spectroscopic data.^{38 (link),45}

Elsayed M.S., Su Y., Wang P., Sethi T., Agama K., Ravji A., Redon C.E., Kiselev E., Horzmann K.A., Freeman J.L., Pommier Y, & Cushman M. (2017). Design and Synthesis of Chlorinated and Fluorinated 7-Azaindenoisoquinolines as Potent Cytotoxic Anticancer Agents That Inhibit Topoisomerase I. Journal of medicinal chemistry, 60(13), 5364-5376.

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Characterization of Synthesized Nanoparticles

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A Varian CARY 100 UV–vis
spectrophotometer (Mulgrave, Victoria, Australia) was used for spectrophotometric
measurements. High performance liquid chromatography (HPLC) instrument
consisting of a Waters 1525 binary HPLC pump, an in-line degasser,
and a Waters 2998 PDA detector (Milford, Massachusetts) was used for
HPLC determinations. In the study, a ZORBAX eclipse C18 (250 ×
4.6 mm, 5 μm) reverse-phase column was used as the stationary
phase. The characterization of the synthesized nanoparticles was performed
by scanning transmission electron microscopy (STEM)–energy-dispersive
X-ray spectrometry (EDS) analysis (Thermo Scientific Quattro FEG SEM),
Fourier transform infrared spectroscopy (FTIR, Agilent Carry 630 FTIR-ATR),
and X-ray photoelectron spectroscopy (XPS) conducted using a K-Alpha
spectrometer (Thermo Fisher) employing a monochromated Al Kα
X-ray source (hν = 14,686.6 eV).

Avan A.N., Demirci-Çekiç S, & Apak R. (2022). Colorimetric Nanobiosensor Design for Determining Oxidase Enzyme Substrates in Food and Biological Samples. ACS Omega, 7(48), 44372-44382.

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