Gel permeation chromatography

Manufactured by Waters Corporation

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Gel permeation chromatography is a technique used to separate and analyze molecules based on their size and molecular weight. It utilizes a porous gel material to allow smaller molecules to penetrate the pores, while larger molecules are excluded, resulting in their separation.

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Lab products found in correlation

Cdcl3, by Bruker (1 mentions) Uv vis spectroscopy, by Agilent Technologies (1 mentions) Nmr spectrometer, by Bruker (1 mentions) Dpx 300, by Bruker (1 mentions) Waters 2487 dual λ absorbance detector, by Waters Corporation (1 mentions) Waters 2414 refractive index detector, by Waters Corporation (1 mentions) Styragel column, by Waters Corporation (1 mentions) Orbital shaker, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Gel permeation chromatography (GPC) system (4 citations) Gel permeation chromatography (GPC (2 citations)

8 protocols using gel permeation chromatography

Synthesis and Characterization of Acrylate-Terminated Polymers

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B and S monomers were reacted at a molar ratio of 1.1:1 at 90°C with stirring overnight to form acrylate-terminated base polymers. Base polymers were then dissolved in anhydrous THF at 167 mg/mL and added to end-capping E monomers (0.5 M in THF) at a 3:2 volume ratio and allowed to react at room temperature with stirring for 1 hour, at which time polymers were washed twice in diethyl ether to remove unreacted monomers and oligomers. Solvents were removed in a vacuum desiccant chamber for 2 days, at which point polymers were dissolved in DMSO at 100 mg/mL, and single-use aliquots were stored at −20°C with desiccant.
Nuclear magnetic resonance spectroscopy (NMR) was used to characterize polymer structure via ¹H NMR in CDCl₃ (Bruker 500 MHz) and analyzed using TopSpin 3.5 software (Billerica, MA, USA). Gel permeation chromatography (Waters, Milford, MA) measurements were performed to measure polymer molecular weight and polydispersity. Polymers were dissolved in BHT-stabilized tetrahydrofuran and 5% DMSO and 1% piperidine, filtered through a 0.2 μm PTFE filter and measured against linear polystyrene standards.

Choi J., Rui Y., Kim J., Gorelick N., Wilson D.R., Kozielski K., Mangraviti A., Sankey E., Brem H., Tyler B., Green J.J, & Jackson E.M. (2019). Nonviral Polymeric Nanoparticles for Gene Therapy in Pediatric CNS Malignancies. Nanomedicine : nanotechnology, biology, and medicine, 23, 102115.

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In Vivo Degradation Study of PCL Graft

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All animals (Sprague Dawley [SD] rats, male, 7 weeks, 200–300 g) were cared for according to methods approved by the Institutional Animal Care and Use Committee (IACUC No. 2020-0019 and 2021-0010) at SMG-SNU Boramae Medical Center, Seoul, Republic of Korea.
The degradation pattern of the PCL graft in the body was observed. After being subcutaneously implanted in the SD rats, the graft was harvested at 3 months, 6 months, 9 months, and 12 months to observe the appearance and microstructure. In addition, the molecular weight and tensile strength at 12 months were measured by gel permeation chromatography (Waters) and universal testing machine, respectively.

Kim S.H., Park S.J., Xu B., Lee J.H., An S.J, & Cha M. (2022). Development of polycaprolactone grafts with improved physical properties and body stability using a screw extrusion-type 3D bioprinter. International Journal of Bioprinting, 9(2), 652.

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Extraction and Analysis of Birch, Beech, and Pine Lignin

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Birch lignin was extracted according to a literature procedure36 . In a 1-litre round-bottom flask with condenser, 40 g of pre-ground birch wood and 400 ml methanol containing 3% hydrogen chloride by weight were combined. The mixture was refluxed for 12 h under stirring, and cooled to room temperature. Residue was removed by suction filtration and washed with additional small portions of methanol. The filtrate was concentrated to <200 ml by rotary evaporation and then poured into 1-litre of ice-cold water with vigorous stirring, causing a light brown solid to precipitate. This lignin was collected by filtration, washed with a small portion of water and dried under vacuum. The yield of dried, crude birch lignin was 4.97 g (12.4 wt%). Beech lignin and pine lignin were extracted in the same way, and the yield was 4.94 g (12.3 wt%) and 3.68 g (9.2 wt%), respectively. The Gel Permeation Chromatography (Waters 1515) was performed to analyse the molecular weights of the extracted lignin. Before analysis, lignin was dissolved in tetrahydrofuran. A calibration curve was obtained using monodisperse polystyrene standards.

Shao Y., Xia Q., Dong L., Liu X., Han X., Parker S.F., Cheng Y., Daemen L.L., Ramirez-Cuesta A.J., Yang S, & Wang Y. (2017). Selective production of arenes via direct lignin upgrading over a niobium-based catalyst. Nature Communications, 8, 16104.

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Characterization of Conjugated Polymers

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Polystyrene was used as a standard for gel permeation chromatography (Waters, Worcester County, MA, USA) to determine the molecular weights of the obtained CPs. UV-visible (UV-vis) absorption spectra of CP were characterized using UV-vis spectroscopy (Agilent, Santa Clara, CA, USA) in solution (chloroform) and film states, respectively. Ultraviolet photoelectron spectroscopy (UPS, Riken, Tokyo, Japan) was performed to determine the energy levels of the CPs. The surface morphologies of CP thin films were characterized using atomic force microscopy (AFM, Park Systems, Suwon, Republic of Korea) in a non-contact mode. The degree of CP chain assembly in the film state was analyzed using 2D-GIXRD (Xenocs, Grenoble, France), and the atomic binding energies of the CPs were determined using X-ray photoelectron spectroscopy (XPS, Thermo-Fisher, Seoul, Republic of Korea). All measurements using the CP thin films were performed with the same thickness (57.0 ± 5.0 nm) applied to the gas sensor device.

Ahn Y., Hwang S., Kye H., Kim M.S., Lee W.H, & Kim B.G. (2023). Side-Chain-Assisted Transition of Conjugated Polymers from a Semiconductor to Conductor and Comparison of Their NO2 Sensing Characteristics. Materials, 16(7), 2877.

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Polymer Characterization by Spectroscopy

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All chemical structures were measured by a 400 MHz nuclear magnetic resonance (NMR) spectrometer (Bruker, Karlsruhe, Germany). The number-average molar mass (M_n,GPC) and Đ were determined by a gel permeation chromatography (Waters, Milford, MA, USA) using DMF (1.0 mL/min) as the eluent with polystyrene standards. The absorption spectrum was measured by UV-Visible Spectroscopy (Sahimadzu, Kyoto, Japan).

Tao H., Xia L., Chen G., Zeng T., Nie X., Zhang Z, & You Y. (2019). PET-RAFT Polymerization Catalyzed by Small Organic Molecule under Green Light Irradiation. Polymers, 11(5), 892.

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Synthesis and Characterization of PLA, PCL, and PLCL

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High molecular weight poly(L-lactide) (PLA), poly(ε-caprolactone) (PCL) and poly(L-lactide-co-ε-caprolactone) (PLCL) were synthesized by ring-opening polymerization or co-polymerization of the corresponding monomers (L-lactide, ε-caprolactone) in the presence of Tin(II) 2-ethylhexanoate as a catalyst in bulk as described previously [27] (link), [28] . The molecular weight of the polymers was determined using a gel permeation chromatography (Waters Corporation) and the details are as follows: PLA: M_w = 220 000 g/mol and M_n = 98 000 g/mol; PCL: M_w = 120 000 g/mol and M_n = 80 000 g/mol; PLCL300: M_w = 316 000 \g/mol and M_n = 120 000 g/mol; PLCL150: M_w = 162 000 g/mol and M_n = 57 000 g/mol. The content of ε-caprolactone in both PLCL copolymers was found to be 7% mol as analyzed by the ¹H nuclear magnetic resonance analysis (DPX 300, Bruker).

Kasoju N., Kubies D., Kumorek M.M., Kříž J., Fábryová E., Machová L., Kovářová J, & Rypáček F. (2014). Dip TIPS as a Facile and Versatile Method for Fabrication of Polymer Foams with Controlled Shape, Size and Pore Architecture for Bioengineering Applications. PLoS ONE, 9(10), e108792.

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Indirect Molecular Weight Determination

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The molecular weight of grafted brushes on the surface could not be directly measured. To learn about the chain length information, an indirect way was used to evaluate the molecular weight of the brushes under the same conditions. An appropriate amount of initiator was put into the solution and polymerization was initiated. Waters gel permeation chromatography (GPC) system equipped with a set of Waters Styragel columns was used to perform the GPC measurements. Both Waters-2414 refractive index detector and Waters-2487 dual λ absorbance detector were used in the columns. The diluent was HPLC grade THF, and the flow rate was low (1.0 mL/min). A series of polystyrene standards with near-monodisperse was used for the calibration.

Wang B., Ye Z., Tang Y., Han Y., Lin Q., Liu H., Chen H, & Nan K. (2016). Fabrication of nonfouling, bactericidal, and bacteria corpse release multifunctional surface through surface-initiated RAFT polymerization. International Journal of Nanomedicine, 12, 111-125.

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Monitoring PLGA-CMPs Release Kinetics

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5 mL of the F_v, F_VR, and F_MR-VR microspheres suspension in the release medium (100 mg/mL in 1X PBS, pH 7.4) was incubated in an orbital shaker (Thermo Scientific). At predetermined time points, the microspheres were collected and weighted. Then, the samples were lyophilized and weighted again. The water uptake was calculated by subtracting the values as mentioned earlier.
The gel permeation chromatography (Waters, USA) was used to study the decrease of PLGA-CMPs molecular weight during the release time. Tetrahydrofuran (THF) was used mobile phase (flow rate of 1.5 ml/min). The Breeze GPC software was applied to calculate the molecular weights of the samples.

Omidi M., Mansouri V., Amirabad L.M, & Tayebi L. (2021). Impact of Lipid/Magnesium Hydroxide Hybrid Nanoparticles on Stability of Vascular Endothelial Growth Factor-Loaded PLGA Microspheres. ACS applied materials & interfaces, 13(21), 24370-24384.

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