Pure methanol

Manufactured by Merck Group

Sourced in Germany, United States

Pure methanol is a colorless, volatile, and flammable liquid. It is primarily used as a solvent in various laboratory applications, including analytical chemistry, biochemistry, and organic synthesis. Methanol serves as a key component in many standard laboratory procedures and experiments.

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Lab products found in correlation

Chloroquine, by Merck Group (2 mentions) Acetonitrile, by Merck Group (2 mentions) Potassium ferricyanide, by Merck Group (2 mentions) Giemsa azur eosin methylene blue solution, by Merck Group (2 mentions) Methanol, by Merck Group (2 mentions) 2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (2 mentions) Citrate buffer, by Carl Roth (1 mentions) Oasis hlb spe column, by Waters Corporation (1 mentions) Ethyl acetate, by Sinopharm (1 mentions) N hexane, by Sinopharm (1 mentions) Acetic anhydride, by Sinopharm (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Dapi blue, by Merck Group (1 mentions) Methanol, by Eppendorf (1 mentions) Colorimetric kit, by Merck Group (1 mentions) Glibenclamide, by Merck Group (1 mentions) Apigenin, by Merck Group (1 mentions) Rosmarinic acid, by Merck Group (1 mentions) Coumaric acid, by Merck Group (1 mentions) Aluminum trichloride alcl3, by Merck Group (1 mentions)

16 protocols using pure methanol

DPPH Assay for Antioxidant Capacity

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The total antioxidant activity (TAA %) of extracts and EOs was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method [23] . Pure methanol (Sigma-Aldrich) was used to calibrate the spectrophotometer. An aliquot of 2 mL of stock solution of 0.1 mM DPPH (Sigma-Aldrich) reagent dissolved in Pure methanol was added to a test tube containing 2 mL of the sample solution in methanol (200 lg/L). The mixture was mixed for approximately 10 s and left to stand in fibber box at room temperature in the dark for 30 min. The absorbance was measured at 517 nm, using a UV scanning spectrophotometer (Unico Ò 1200).
TAA % was expressed as the percentage inhibition of the DPPH radical using the following equation: TAA (%) = (A 0 -A s /A 0 ) 9 100, where TAA is the total antioxidant activity, A 0 (control) is the absorbance of DPPH solution in methanol and As is the absorbance of a DPPH solution with the tested sample (essential oils and extracts) or the positive controls (Tannic acid and (?)catechin) solutions. The control contained 2 mL of DPPH solution and 2 mL of methanol. The average values of TAA % were carried out for three replicates, and are expressed as mean values ± standard deviation (SD). Also, the antioxidant activity of each extract was expressed in terms of IC 50 (the concentration required to inhibit DPPH radical formation by 50 %), calculated from the inhibition curve [24] .

Salem M.Z.M., Zayed M.Z., Ali H.M, & Abd El-Kareem M.S.M. (2016). Chemical composition, antioxidant and antibacterial activities of extracts from Schinus molle wood branch growing in Egypt. J Wood Sci., 62(6).

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Lichenysin Extraction from Liquid Cultures

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Lichenysin was extracted as described by Madslien et al. (2013) (link) and Rønning et al. (2015) (link) with slight modifications. Briefly, a 5 ml aliquot of the collected liquid cultures (see section “Growth Curves in LB Broth and Skimmed Milk”) was mixed with 5 mL of pure methanol (Merck) in a soda-lime glass tube (DWK Life Sciences), and vortexed for 5 min until a homogenous mixture was obtained. The mixture was heated for 30 min at 80°C in a water bath. At 10 min intervals, each tube was vortexed for 20 s. Subsequently, the mixture of methanol and cells was cooled down for 20 min on the bench, and centrifuged for 10 min at 4000 g at room temperature. The liquid phase was transferred to a new glass tube. The cell pellet was subsequently resuspended in 2 ml of methanol, and the extraction process was repeated. Per sample, the liquid phases collected from the first and second extraction procedures were mixed and evaporated under N₂ flow at 40°C in a heating block. The dry residue was resuspended in 500 μl of methanol and lichenysin was quantified in these samples.

Yeak K.Y., Perko M., Staring G., Fernandez-Ciruelos B.M., Wells J.M., Abee T, & Wells-Bennik M.H. (2022). Lichenysin Production by Bacillus licheniformis Food Isolates and Toxicity to Human Cells. Frontiers in Microbiology, 13, 831033.

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Antimalarial Drug Potency Assessment

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Chloroquine was purchased from Sigma-Aldrich. Dihydroartemisinin and piperaquine were kindly provided by Mangalam Drugs and Organics Ltd., Mumbai, India. Stock solutions of Chloroquine and piperaquine were prepared in distilled water, dihydroartemisinin in pure methanol (Merck, Darmstadt, Germany). Drug concentration ranges were chosen based on the average IC50 values available in the literature^{10 (link),29 (link)–33 (link),35 ,51 –54 (link)}. Doubling concentrations ranging from 6 to 100 nM for Chloroquine, and 6 to 486 nM for piperaquine, 0.12 to 32 nM for dihydroartemisinin were tested.

Molnár P., Orbán Á., Izrael R., Babai R., Marton L., Butykai Á., Karl S., Vértessy B.G, & Kézsmárki I. (2020). Rapid and quantitative antimalarial drug efficacy testing via the magneto-optical detection of hemozoin. Scientific Reports, 10, 14025.

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Immunohistochemical Detection of CTGF

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After dewaxing, brain sections were boiled (in an 850 W microwave oven) for 15 min in citrate buffer (2.1 g citric acid monohydrate/l, pH = 6) (Carl Roth, Karlsruhe, Germany). Endogenous peroxidase was inhibited by 1% H₂O₂ in pure methanol (Merck, Darmstadt, Germany) for 15 min. Sections were incubated with 10% normal pig serum (Biochrom, Berlin, Germany) to block nonspecific binding of immunoglobulins and then with the polyclonal antibody CTGF (1:100, BMA; USbio, Swampscott, Massachusetts, USA) overnight at 4°C. Antibody binding to tissue sections was visualized with a biotinylated rabbit anti-mouse IgG F(ab)₂ antibody fragment (1 : 400; Dako, Hamburg, Germany). Subsequently, sections were incubated with a horseradish peroxidase-conjugated streptavidin complex for 30 min (1: 100; Dako), followed by development with diaminobenzidine substrate (Fluka, Neu-Ulm, Germany). Finally, sections were counterstained with Mayer’s hemalaun. As negative controls, the primary antibodies were excluded.

Liu Y., Liu Z., Li X., Luo B., Xiong J., Gan W., Jiang M., Zhang Z., Schluesener H.J, & Zhang Z. (2014). Accumulation of connective tissue growth factor+ cells during the early phase of rat traumatic brain injury. Diagnostic Pathology, 9, 141.

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Decoquinate Compound Characterization Protocol

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A decoquinate (CAS: 18507-89-6, purity ≥ 98%) standard was obtained from Dr. Ehrenstorfer GmbH (Augsburg, Germany). Chromatographically, pure methanol and acetonitrile were purchased from Merck (Mumbai, India). Ethyl acetate, acetic anhydride, acetic acid, n-hexane and trichloromethane were all provided by Sinopharm Chemical Reagents Co., Ltd. (Shanghai, China). Pyridine and nylon needle filters (organic phase, 0.22 µm) were obtained from Shanghai Macklin Biochemical Technology Co., Ltd., Shanghai, China and Anpel Experimental Technology Co., Ltd., Shanghai, China respectively. An Oasis HLB SPE column (column capacity of 3 mL) was purchased from Waters Company. The ultrapure water used in the experiment was made in the laboratory (UK Kertone Co., Ltd., Daventry, UK).

Zhu Y., Chen L., Guo Y., Gao P., Liu S., Zhang T., Zhang G, & Xie K. (2023). Quantitative Analysis of Decoquinate Residues in Hen Eggs through Derivatization-Gas Chromatography Tandem Mass Spectrometry. Foods, 13(1), 119.

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Parasite Load Quantification Assay

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For the in vitro experiments parasite load was checked as previously described (32 (link)). Briefly, after each time point supernatant was collected and reserved. The plates were fixed with pure methanol (Merck) or 4% paraformaldehyde (Sigma Aldrich) diluted in PBS for at least 15 min at room temperature. Then the wells were gently washed with warm PBS and incubated with 5 μg/mL DAPI (blue) (Sigma Aldrich) diluted in PBS. At least 6 images of each well were acquired on the IN-Cell Analyzer 2200 equipment, counted, and the average per well was graphically plotted.

Amaral M.P., Cardoso F.D., de Farias I.S., de Souza R.Q., Matteucci K.C., Torrecilhas A.C, & Bortoluci K.R. (2023). NAIP/NLRC4 inflammasome participates in macrophage responses to Trypanosoma cruzi by a mechanism that relies on cathepsin-dependent caspase-1 cleavage. Frontiers in Immunology, 14, 1282856.

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Intracellular Metabolite Extraction Protocol

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Cells were washed with ice-cold PBS. After trypsin digestion and centrifugation, detached cells were collected and then immediately stored at − 80 °C until analysis. Stored samples were thawed slowly on ice. To extract intracellular metabolites, 500 µL of pure methanol (Merck, Germany) was added to the sample. The mixture was vortexed for 5 min (Jingxin MIX-200, Shanghai, China), and subjected to three freeze‒thaw cycles. After centrifugation at 4 °C for 10 min (Eppendorf 5427R, Germany), 300 µL of methanol containing the internal standard was added to the sample. The mixture was then vortexed for one min and placed in a − 20 °C freezer for one hr. Then, the sample was centrifuged at 12,000 rpm at 4 °C for 10 min. 200 µL of the supernatant was retained and allowed to stand for 30 min. Then, centrifugation was repeated under the same conditions. The collected supernatant was then stored in a sample injection bottle for further analysis.

Li F., Huang H., Xu J., Tao L., Zhou L., Hsueh C., Gong H, & Zhang M. (2023). Fusobacterium nucleatum-triggered purine metabolic reprogramming drives tumorigenesis in head and neck carcinoma. Discover. Oncology, 14, 120.

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Analytical Techniques for Pharmaceutical Assays

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The solvents and chemicals were used to maintain the analytical and laboratory-grade (e.g., SIGMA, E. Merck, or BDH) solvents and chemicals were used in most of the experiments. Whitman No.1 (Sargent-Welch, USA), Tween-80, Loperamide (Square Pharmaceuticals Ltd., Bangladesh), Streptokinase (Skinase, Popular Pharmaceutical Ltd., Bangladesh), Pure methanol (Merck KGaA, Darmstadt, Germany), Potassium ferricyanide (1%), Phosphate buffer saline, 0.1% ferric chloride, DMSO solution, and NBT (Nitroblue-tetrazolium).

Islam M.R., Proma N.M., Naima J., Uddin M.G., Afrin S.R., & Hossain M.K. (2020). Assessment of the Pharmacological Activities of Ardisia solanacea Roxb: An Ethnomedicinal Plant used in Bangladesh. Journal of Pharmacy and Nutrition Sciences, 10(5), 302-314.

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Measurement of Pollutants and Micropollutants in Wastewater

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In all batch and continuous-flow samples, concentrations of conventional pollutants, i.e. NO 3 -N, NO 2 -N, NH 4 -N, total and soluble COD (sCOD) were measured with Hach-Lange/Merck colorimetric kits followed by spectrophotometry, as previously described in Polesel et al. (2017) . Wastewater fractionation and quantification of the organic carbon availability (mg L -1 of biodegradable COD with its two sub-fractions, i.e., readily (Ss) and slowly (Xs) biodegradable COD) was performed according to Roeleveld and Van Loosdrecht (2002) by combination with BOD-analysis and measure of total and soluble COD. Samples for micropollutants were analyzed using HPLC-MS/MS as described in Escolà Casas et al. (2015) and in Polesel et al., (2017) . Briefly, 4 mL of aqueous samples were sampled and
13 stored in glass vials with addition of 1.4 mL pure methanol (99.9%, Merck Millipore). Samples were frozen at -20 o C prior analysis. For the analysis 1.5 mL of each sample were transferred to an HPLC vial and centrifuged (6000 rpm, 10 min). Subsequently, 100 µL of internal standard solution was added to 900 mL of the centrifuged samples solution. Samples were analyzed with an injected volume of 100 µL. Information regarding targeted micropollutants, HPLC-MS/MS and mass spectrometry conditions, limit of quantification and detection are shown in Escolà Casas et al. (2015) .

Torresi E., Gülay A., Polesel F., Jensen M.M., Christensson M., Smets B.F, & Plósz B.G. (2018). Reactor staging influences microbial community composition and diversity of denitrifying MBBRs- Implications on pharmaceutical removal. Water research, 138.

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Comprehensive Chemical Inventory for Scientific Research

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All the required chemicals including chloride (FeCl₃), Sodium hydroxide (NaOH), nitric acid (HNO₃), iron hydrochloric acid, Folin-Ciocalteu, sodium carbonate (Na₂CO₃), glibenclamide, 2, 2-diphenyl 1-picrylhydrazyl, potassium ferricyanide, phosphate buffer, and aluminum trichloride (AlCl₃) were procured from the Sigma Aldrich Company based in St. Louis, MO, USA. Analytical grade chemicals and reagents were utilized in this study except for pure m ethanol, ethanol, quercetin, vanillin, hydrochloric acid, acetic acid, acetonitrile, tannic acid, caffeine acid, gallic acid, ascorbic acid, rosmarinic acid, coumaric acid, quercetin, rutin, apigenin, catechin, catechol, naringenin, L-Hystidine, Mueller-Hinton, Sabouraud gel, and D-glucose, which were procured from Sigma-Aldrich in Steinheim, Germany.

Zahrae Radi F., Bencheikh N., Anarghou H., Bouhrim M., Alqahtani A.S., Hawwal M.F., Noman O.M., Bnouham M, & Zair T. (2023). Quality control, phytochemical profile, and biological activities of Crataegus monogyna Jacq. and Crataegus laciniata Ucria fruits aqueous extracts. Saudi Pharmaceutical Journal : SPJ, 31(10), 101753.

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