Extracellular polysaccharides and proteins (EPS) from cellulose acetate membrane coupons-attached biofilms were detached through ultrasonication in 10 mL 0.9% NaCl, as mentioned in the previous section. After ultrasonication, the suspension harboring detached cells and EPS from CA membrane coupons was centrifuged (8,000 × g, 10 min). 0.25 μm syringe-filtered cell-free supernatant was used for quantification of EPS using liquid chromatography with organic carbon detector (LC-OCD) model-8 (DOC-Labor, Germany) equipped with a Toyopearl size exclusion chromatography column TSK HW50S (Tosoh, Japan) (Dimension: 250 mm × 20 mm, particle size: 20–40 μm). Polysaccharides and proteins from total EPS were fractionated and resolved using organic carbon detector (OCD) and organic nitrogen detector (OND). ChromCALC uni software was used to determine the concentration of each fraction in organic matter below the curve based on the integration of the defined area (Huber et al., 2011 (link); Stewart et al., 2013 (link)).
Tsk hw50s
Manufactured by Tosoh
Sourced in Japan
The TSK HW50S is a size exclusion chromatography (SEC) column used for the separation and analysis of macromolecules. It features a porous gel-type stationary phase designed to fractionate samples based on their molecular size or hydrodynamic volume.
Automatically generated - may contain errors
2 protocols using tsk hw50s
1
Extracellular Polysaccharides and Proteins Quantification
2
Comprehensive Water Quality Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Standard Methods65 were used to evaluate turbidity (Method 2130B;
Hach 2100 N turbidimeter, Loveland, CO), pH (4500-H+B Electrometric
method; Orion 720A pH meter, Thermo Fisher Scientific, Waltham, MA),
alkalinity (Method 2320; titration method with pH end point of 4.5),
DOC concentration (filtration through prerinsed 0.45 μm Nylaflo
membranes, Pall, Port Washington, NY; Method 5310B; Shimadzu TOC-V
CPH analyzer, Kyoto, Japan) with a reporting limit of 0.2 mg/L, and
ultraviolet absorbance at 254 nm (UVA254; Method 5910B;
1 cm quartz cell; Hach DR 5000 Spectrophotometer, Loveland, CO). Specific
ultraviolet absorbance at 254 nm (SUVA) was calculated by dividing
UVA254 absorbance by the DOC concentration.69 (link)Liquid chromatography in combination with
organic carbon detection (LC-OCD) was used to fractionate DOC (as
biopolymers [BPs], humic substances [HS], building blocks [BB], low
molecular-weight [LMW] neutrals, LMW acids) as described in Huber
et al.66 (link) Samples were first filtered through
a prerinsed 0.45 μm polyether sulfone membrane (Millipore Express
PLUS; Merck Millipore, Burlington, MA). Chromatographic separation
was completed using a weak cationic exchange column (Toyopearl, TSK
HW 50S, Tosoh, Japan).
Hach 2100 N turbidimeter, Loveland, CO), pH (4500-H+B Electrometric
method; Orion 720A pH meter, Thermo Fisher Scientific, Waltham, MA),
alkalinity (Method 2320; titration method with pH end point of 4.5),
DOC concentration (filtration through prerinsed 0.45 μm Nylaflo
membranes, Pall, Port Washington, NY; Method 5310B; Shimadzu TOC-V
CPH analyzer, Kyoto, Japan) with a reporting limit of 0.2 mg/L, and
ultraviolet absorbance at 254 nm (UVA254; Method 5910B;
1 cm quartz cell; Hach DR 5000 Spectrophotometer, Loveland, CO). Specific
ultraviolet absorbance at 254 nm (SUVA) was calculated by dividing
UVA254 absorbance by the DOC concentration.69 (link)Liquid chromatography in combination with
organic carbon detection (LC-OCD) was used to fractionate DOC (as
biopolymers [BPs], humic substances [HS], building blocks [BB], low
molecular-weight [LMW] neutrals, LMW acids) as described in Huber
et al.66 (link) Samples were first filtered through
a prerinsed 0.45 μm polyether sulfone membrane (Millipore Express
PLUS; Merck Millipore, Burlington, MA). Chromatographic separation
was completed using a weak cationic exchange column (Toyopearl, TSK
HW 50S, Tosoh, Japan).
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