Dry milled corn bran was gifted from Agricor, Ltd. (Marion, IN, USA). A commercial sample of low methoxy pectin from the citrus peel (900-69-5) and fructooligosaccharide (FOS—F8052) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Thermostable α-amylase, laccase from Trametes versicolor, and methanolic-HCl were obtained from Sigma-Aldrich. Hexane, sodium hydroxide pellet, ethanol, concentrated HCl, and Tri-Sil were purchased from Fisher Scientific (Thermo Fisher Scientific, Suwanee, GA, USA).
Methanolic hcl
Manufactured by Merck Group
Sourced in United States
Methanolic HCl is a reagent primarily used in analytical chemistry. It is a solution of hydrogen chloride (HCl) in methanol (CH3OH). This reagent is commonly utilized for the derivatization of carboxylic acids, alcohols, and other functional groups, facilitating their analysis and identification.
Automatically generated - may contain errors
Lab products found in correlation
Hexane, by Thermo Fisher Scientific (1 mentions)
Ethanol, by Thermo Fisher Scientific (1 mentions)
Thermostable α amylase, by Merck Group (1 mentions)
Laccase from trametes versicolor, by Merck Group (1 mentions)
Concentrated hcl, by Thermo Fisher Scientific (1 mentions)
Tripentadecanoin, by Merck Group (1 mentions)
Bradford protein assay, by Thermo Fisher Scientific (1 mentions)
Sodium hyaluronate, by Merck Group (1 mentions)
N acetyl d galactosamine, by Merck Group (1 mentions)
L fucose, by Merck Group (1 mentions)
D glucosamine, by Merck Group (1 mentions)
D mannose, by Merck Group (1 mentions)
D galactose, by Merck Group (1 mentions)
D galactosamine, by Merck Group (1 mentions)
N acetyl d glucosamine, by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
N hexane, by Merck Group (1 mentions)
Db 5ms fused silica capillary column, by Agilent Technologies (1 mentions)
Gc fid 7890a, by Agilent Technologies (1 mentions)
Silica gel 60, by Merck Group (1 mentions)
10 protocols using methanolic hcl
1
Corn Bran Valorization and Pectin Analysis
2
Fatty Acid Profiling in L. infantum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1 M methanolic HCl (500 mL) (Sigma–Aldrich) and 10 μL of 5 mg mL−1 tripentadecanoin (Sigma–Aldrich) as internal standard were added to L. infantum cell pellets (1 × 108 cells), followed by transmethylation of fatty acids at 85 °C for 4 h. The fatty acid methyl esters (FAMEs) were partitioned into 200 μL hexane from the aqueous phase by the addition of 0.9% KCl. FAMEs were analysed by GC-FID according to the method described in Larson and Graham (2001).39 (link)
3
Fatty Acid Analysis of Drosophila
4
Monosaccharide Composition Analysis by GC-MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The monosaccharide composition analysis was determined by GC–MS28 (link). ESM-C (approximately 0.5 mg) was methanolyzed with 0.5 M methanolic/HCl (Sigma–Aldrich, USA) at 84 °C for 16 h. After cooling, a solution containing 500 μL methanol, 50 μL acetic acid, and 10 μL pyridine was added and vortexed for 30 min at room temperature. After evaporation to dryness under a stream of N2 gas, the reactants were treated with Sylon HTP (HMDS/TMCS/pyridine, 2:1:10) trimethylsilyation reagent (Supelco, PA) at 85 °C for 30 min. The final trimethylsilyated (TMS) derivatives were dissolved in 400 μL n-hexane (GC grade, Supelco, PA, USA) for analysis on a Bruker SCION SQ GC–MS instrument (Bruker, Billerica, USA), which was fitted with a DB-5MS fused silica capillary column (length 30 m, inner diameter 0.25 mm, Agilent J & W Scientific, USA). Sodium hyaluronate, L-fucose, D-glucose, D-galactose, D-mannose, D-glucosamine, D-galactosamine, N-acetyl-D-glucosamine, and N-acetyl-D-galactosamine (all Sigma) were also treated with the above derivatization reactions and used as standards.
5
Fatty Acid Extraction and Characterization in Drosophila
6
Monosaccharide Profiling of Onion Peels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SDS washed and de-starched dry onion peels were weighed and added to a 2 mL screw -top microcentrifuge tube and 1 mL of 3 M HCl in methanol (Sigma, USA) was added to the walls and incubate for 16 h at 80°C in heating block in a fume hood as per De Ruiter et al, 1992. The tubes were cooled on ice, briefly spin down to collect condensate, and evaporated under a gentle stream of airflow at 25°C. One mL of 2 M TFA stock (Sigma, USA) was added to the methanolic HCl-treated walls and incubated for 1 h at 120°C, cooled down on ice, and centrifuged at 8,000 g for 5 min to pellet any solids. The supernatant from the met/TFA hydrolysis was separated from the remaining residue using a pipette and both fractions were saved in separate tubes.
Both tubes had any remaining liquid evaporated under a gentle stream of airflow at 25°C. The dried soluble supernatant was then resuspended in 1 mL ddH20 and filtered through a 0.2 µm nylon filter for anion exchange chromatography. The insoluble residue had a small amount of water added to the tube, frozen in -80°C, and lyophilized overnight. To correct for monosaccharide losses due to degradation during the met/TFA hydrolysis, the µg per mg amounts for each monosaccharide (Tables 1234) had correction factors applied according to (35) : Glc (0.94), GalA (0.71), Gal (0.86), Xyl (0.91), Rha (0.83), Ara (0.83), Fuc (0.93), GlcA (0.63), Man (0.89).
Both tubes had any remaining liquid evaporated under a gentle stream of airflow at 25°C. The dried soluble supernatant was then resuspended in 1 mL ddH20 and filtered through a 0.2 µm nylon filter for anion exchange chromatography. The insoluble residue had a small amount of water added to the tube, frozen in -80°C, and lyophilized overnight. To correct for monosaccharide losses due to degradation during the met/TFA hydrolysis, the µg per mg amounts for each monosaccharide (Tables 1234) had correction factors applied according to (35) : Glc (0.94), GalA (0.71), Gal (0.86), Xyl (0.91), Rha (0.83), Ara (0.83), Fuc (0.93), GlcA (0.63), Man (0.89).
7
Derivatization of Fatty Acids from Fly Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
200 µl of methanolic HCl (Supelco Analytical, Sigma–Aldrich Co., St. Louis, MO) was added to dried, crude whole fly extract from about 500 flies and incubated for 1.5 to 2 hr at 60°C with occasional vortexing. After the acid-based catalysis, the reaction was cooled on ice, followed by the addition of 50 µl of water and 50 µl of hexane, and brief vortexing. The hexane layer (which contains the fatty acid methyl esters) was removed for GCMS analysis. Concurrently, synthetic standards containing 5 carbons (tiglic acid, trans-2-pentenoic acid, trans-3-pentenoic acid, and 3-methyl crotonic acid [TCI Chemicals Co., Tokyo, Japan]) were treated with the same reactions. Methyl angelate (TCI Chemicals Co.) was not treated.
8
Fatty Acid Quantification in HepG2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fatty acid levels (µg) were normalized per milligram protein, quantified by Bradford assay [29] , in HepG2 pellets. Pellets were dissolved in 0.2 mL cell lysis buffer containing 1mM phenylmethanesulfonyl fluoride [30] , followed by sonication, to obtain cell lysate. Total lipids extraction [28] was achieved after adding 0.05mg pentadecanoic acid (internal standard) to cell pellets. After drying, synthesis of fatty acid methyl esters (FAME) using 3mol/l methanolic HCl (Supelco, Bellafonte, PA, EEUU) for 1-hour at 90°C, was performed. FAME were analysed by gas-chromatography [31] and fatty acid concentrations were determined in relation to peak area of internal standard.
9
Lipid Profiling of Mature Brown Rice Flour
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mature brown rice grains were obtained by manual de-hulling and ground with a CapMixTM capsule mixing device (3 M ESPE, Seefeld, Germany). Total lipids from ~300 mg above prepared rice flour samples were extracted with a mixture of chloroform/methanol/0.1 M KCl (at a ratio of 2/1/1, by volume). Fatty acid methyl esters (FAME) were prepared by incubating lipid samples in 1 N Methanolic-HCl (Supelco, Bellefonte, PA) at 80 °C for 2 h. TAG and polar membrane lipid pools were fractionated from total lipids in thin layer chromatography (TLC) (Silica gel 60, Merck, Darmstadt, Germany) using a solvent mixture of hexane/diethylether/acetic acid (at a ratio of 70/30/1, by volume) and individual membrane lipid classes were separated by TLC using a solvent mixture of chloroform/methanol/acetic acid/water (90/15/10/3, by volume). Authentic lipid standards were loaded and were run in separate lanes on the same plates for identification of lipid classes. Silica bands, containing individual class of lipid were used to prepare FAME as mentioned above and were analysed by gas chromatography GC-FID 7890A (Agilent Technologies, Palo Alto, CA) that was fitted with a 30 m BPX70 column (SGE, Austin, TX) for quantifying individual fatty acids on the basis of peak area of the known amount of heptadecanoin that was added in as an internal standard [50 (link)].
10
Structural Characterization of Sulfated Glycans
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!