The PHA content was measured using the spectrometric method as suggested in Law and Slepecky (1960a (link)). The PHAs of the samples were extracted from the cells with dichloromethane (Carl Roth, Karlsruhe, Germany). Two milliliters of the chlorinated hydrocarbon is added to each previously lyophilized sample and the mixture was boiled in a water bath for 10 min. After cooling down to room temperature, cell pellets were centrifuged for 10 min at 3200×g and 4 °C (Allegra X-12R, Beckman Coulter, Brea, USA) and the supernatants were transferred to a 10-mL glass tube. Afterwards, the extraction step was repeated two times. The supernatants were then evaporated to obtain the extracted PHAs. After complete evaporation of the dichloromethane, samples were boiled with 2 mL of concentrated sulfuric acid (98%, Carl Roth, Kalrsruhe, Germany) in a water bath for 30 min. After cooling down to room temperature, 200 μL of the sample was transferred in a microtiter plate and measured in a spectrometer (Eon, BioTek, Winooksi, USA). Thereby, a spectrum was recorded between 200 and 800 nm using 5 nm slit. The lamp of the device provides monochromatic light in the visual and UV range. The maximum absorbance at 235 nm can be used to determine the amount of crotonic acid in the sample.
Sulfuric acid
Manufactured by Carl Roth
Sourced in Germany
Sulfuric acid is a highly corrosive, colorless, and dense liquid chemical compound. It is composed of hydrogen, sulfur, and oxygen, with the chemical formula H2SO4. Sulfuric acid is a strong mineral acid that is widely used in various industrial and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Sodium hydroxide (naoh), by Carl Roth (6 mentions)
Acetic acid, by Carl Roth (4 mentions)
Potassium hydroxide, by Carl Roth (4 mentions)
Ethyl acetate, by Merck Group (4 mentions)
Hydrogen peroxide, by Carl Roth (3 mentions)
Sodium carbonate, by Carl Roth (3 mentions)
Methanol, by Avantor (3 mentions)
0.1 n standard hydrochloric acid, by Carl Roth (3 mentions)
Sodium chloride (nacl), by Carl Roth (3 mentions)
Hydrochloric acid (hcl), by Merck Group (3 mentions)
Dichloromethane, by Carl Roth (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Methanol, by Carl Roth (2 mentions)
Milli q system, by Merck Group (2 mentions)
R 100, by Büchi (2 mentions)
4 anisaldehyde, by Carl Roth (2 mentions)
Silica gel 60 f254 aluminium plates, by Merck Group (2 mentions)
N n dimethylacetamide dmac, by Avantor (2 mentions)
Poly 2 6 dimethyl 1 4 phenylene oxide ppo, by Merck Group (2 mentions)
Hydrochloric acid (hcl), by Carl Roth (2 mentions)
39 protocols using sulfuric acid
1
Quantifying Polyhydroxyalkanoates via Spectrometry
2
Synthesis and Deposition of ZIF-8 Thin Films
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Zn(NO3)2·6H2O (98%, Sigma-Aldrich), 2-methylimidazole (99%, Sigma-Aldrich), methanol (99.5%, Roth), ethanol (99.8%, Fisher-Scientific), tetrahydrofurane (99.5%, Roth), dimethylformamide (99.8%, Sigma-Aldrich), dimethylsulfoxide (99.9%, Sigma-Aldrich), toluene (99.9%, Roth), para-methyl anisate (98%, Alfa-Aesar), iodobenzene (98%, Alfa-Aesar), sulfuric acid (96%, Roth) and hydrogen peroxide (35%, Roth) were used without further purification. For 1H-NMR measurements, DMSO-d6 (99.8 atom % D, Acros-Organics) and DCl (20 wt% in D2O, 99 atom % D, euriso-top) were used.
As substrates for the deposition of ZIF-8 we used silicon wafers which were cut into 1 × 1 cm pieces and glass slides cut into 2 × 2.5 cm pieces. All substrates were cleaned with ethanol and then immersed in a fresh mixture of sulfuric acid and hydrogen peroxide (2 : 1) for 15 minutes. Subsequently, they were washed with water, ethanol and methanol. Cleaned substrates were not stored but directly used.
As substrates for the deposition of ZIF-8 we used silicon wafers which were cut into 1 × 1 cm pieces and glass slides cut into 2 × 2.5 cm pieces. All substrates were cleaned with ethanol and then immersed in a fresh mixture of sulfuric acid and hydrogen peroxide (2 : 1) for 15 minutes. Subsequently, they were washed with water, ethanol and methanol. Cleaned substrates were not stored but directly used.
3
Quantification of Metabolite Production in Strain AK17
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Consumption of glucose, mannitol, and production of lactic acid, acetic acid, and ethanol by strain AK17 and the derived mutant strains were analysed by high performance liquid chromatography (HPLC). The samples for HPLC analysis were filtered through a 0.2 µm filter (Phenomenex) prior injection. Glucose, mannitol, acetic acid, lactic acid, and ethanol were quantified using a Dionex 2000 HPLC system (Dionex, Idstein, Germany) with a Rezex ROA-Organic Acid H + (8%, Phenomenex, Aschaffenburg, Germany) and a RI-101 detector (Shodex, München, Germany). Separation was performed at a column temperature of 60 °C with 0,2 mM sulfuric acid (Carl Roth, Karlsruhe, Germany) as eluent at a flow rate of 600 μl/min for 30 min. Quantification was carried out using external standards with HPLC grade (Merck, Darmstadt, Germany; Sigma-Aldrich, St. Louis, USA) and the Chromeleon Evaluation Software version 6.80 (Dionex, Idstein, Germany).
4
Surface Functionalization with APTES
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Sulfuric acid (100%) and hydrogen peroxide (30%) were purchased from Carl Roth (Karlsruhe, Germany). 3-amino-propyltriethoxysilane (99%) (APTES) was purchased from Sigma Aldrich (Darmstadt, Germany). Toluene (99.7%) was purchased from Honeywell (Hamburg, Germany). Milli-Q water with a resistance of 18 MΩcm was produced in an in-house Milli-Q-system from Merck (Darmstadt, Germany).
5
Chemical Characterization of Authentic Standards
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Chemicals were obtained from the following suppliers: acetone (99.8%) from Acros Organics B.V.B.A., sodium carbonate (≥ 99%, water free), sulfuric acid (98%), d -glucose, malt extract, and sodium hydroxide (≥ 98%) from Carl Roth GmbH & Co. KG (Karlsruhe, Germany), d -fructose from Hamburger Zuckergesellschaft mbH (Hamburg, Germany), sucrose from Südzucker (Mannheim, Germany), and sodium hydrogen carbonate (≥ 99.7%) from Th. Geyer GmbH & Co. KG (Renningen, Germany).
Authentic standard compounds were purchased from commercial sources: geraniol (99%), malic acid (> 99%), 2-nonanone (99%), 1-octen-3-ol (98%), 2-phenylethanol (99%), and terpinen-4-ol (97%) from Acros Organics B.V.B.A (Fair Lawn, USA), phenylacetic acid (99%) from Alfa Aesar (Haverhill, USA), acetic acid (100%), citric acid (water free), and oxalic acid-dihydrate from AppliChem GmbH (Darmstadt, Germany),l -tartaric acid (≥ 99.5%), eugenol (pure) from Carl Roth GmbH, linalool (97%), methyl phenylacetate (> 99%), trans-nerolidol (analytical standard), 2-nonanol (99%), and lactic acid (≥ 85%) from Sigma Aldrich (St. Louis, USA), and 3,4-dimethylbenzaldehyde (95%) from TCI Chemicals (Tokyo, Japan).
Authentic standard compounds were purchased from commercial sources: geraniol (99%), malic acid (> 99%), 2-nonanone (99%), 1-octen-3-ol (98%), 2-phenylethanol (99%), and terpinen-4-ol (97%) from Acros Organics B.V.B.A (Fair Lawn, USA), phenylacetic acid (99%) from Alfa Aesar (Haverhill, USA), acetic acid (100%), citric acid (water free), and oxalic acid-dihydrate from AppliChem GmbH (Darmstadt, Germany),
6
Soxhlet Extraction and Chromatographic Fractionation
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Two separate Soxhlet extractions of 40.95 g and 41.36 g of plant material were carried out with 500 mL of ethanol each for 24 h. The native extracts were evaporated to dryness on a rotary evaporator (Büchi R-100, Büchi, Flawil, St. Gallen, Switzerland). A total of 2.63 g of dry extract yielded from the first Soxhlet extraction were separated by open-column chromatography on 25 g of silica (40–63 µm, VWR) with hexane, ethyl acetate and methanol in solvent systems of rising polarity, leading to 67 fractions.
TLC analyses of fractions were carried out on Silica Gel 60 F254 aluminium plates (Merck, Darmstadt, Hessen, Germany) with mobile phase systems hexane/ethyl acetate/methanol = 75/20/5 (v/v) for apolar fractions and hexane/ethyl acetate/methanol = 60/20/20 (v/v) for polar fractions. The staining reagent consisted of 0.5 mL of 4-anisaldehyde, 10 mL of acetic acid (both Carl Roth), 85 mL of methanol and 5 mL of sulfuric acid (Carl Roth). After spraying, the plates were heated at 105 °C for 5 min.
TLC analyses of fractions were carried out on Silica Gel 60 F254 aluminium plates (Merck, Darmstadt, Hessen, Germany) with mobile phase systems hexane/ethyl acetate/methanol = 75/20/5 (v/v) for apolar fractions and hexane/ethyl acetate/methanol = 60/20/20 (v/v) for polar fractions. The staining reagent consisted of 0.5 mL of 4-anisaldehyde, 10 mL of acetic acid (both Carl Roth), 85 mL of methanol and 5 mL of sulfuric acid (Carl Roth). After spraying, the plates were heated at 105 °C for 5 min.
7
Extraction and Characterization of Natural Bark Compounds
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Fresh wood bark from larch (Larix decidua), spruce (Picea abies) and beech (Fagus sylvatica) was kindly provided from local sawmills in Kuchl (Austria), which was manually debarked and collected. Reagents, such as sulfuric acid (96%), ethanol (99%), 1,4-dioxan, sodium carbonate, tetrahydrofuran were purchased from Carl Roth (Karlsruhe, Germany). Methanol and dimethyl sulfoxide were obtained from VWR (Rue Carnot, France). Folin–Ciocalteau reagent and hydrochloric acid (32%) were obtained from Merck (Darmstadt, Germany). All chemicals were used as received without further purification.
8
Synthesis of Polymeric Membrane Materials
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All chemicals were used without further purification. Poly(2,6-dimethyl-1,4-phenylene oxide) (PPO) was purchased from Sigma Aldrich (Munich, Germany), and bromination was conducted in the same way as described previously [28 (link)]. Poly[(1-(4,4′-diphenylether)-5-oxybenzimidazole)-benzimidazole] (PBI-OO) was purchased from FuMA-Tech GmbH (Ludwigsburg, Germany). Sulfonated polymer was synthesized as previously reported in the literature [29 (link)]. 1,2,4,5-tetramethylimidazole was purchased from TCI chemicals. N,N-Dimethylacetamide (DMAc) and methanol were obtained from VWR International GmbH (Bruchsal, Germany). Sulfuric acid, potassium hydroxide, 0.1 N standard hydrochloric acid, and sodium hydroxide were purchased from Carl Roth GmbH (Karlsruhe, Germany). Vanadium electrolyte was provided by RIVA GmbH Batteries. The structures used in this study are presented in Figure 1 .
9
Quantitative Analysis of Volatile Fatty Acids
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For determination of total volatile fatty acids (VFA) content and ratio of VFAs and alkalinity (VFA/TAC), samples were centrifuged for 15 min and 4,696 × g at 4 °C. 10 mL of the liquid phase were then titrated with 0.05 M sulfuric acid (Carl Roth GmbH, Karlsruhe, Germany) to pH values of 5.00, 4.40, 4.30 and 4.00. VFA as well as VFA/TAC were measured in triplicates and calculated as described elsewhere46 . Composition of VFA was analyzed by atres Analytik (München, Germany) using an in-house gaschromatographic approach. Contents of following VFAs were measured: acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid, hexanoic acid and heptanoic acid.
For analyzing elemental composition (C, N, S) samples were dried at 50 °C for 48 h. Afterwards samples were grinded using mortar and pestle. 10 mg of each sample were oxidized in the combustion tube of the elemental analyzer Vario EL II (Elementar Analysensysteme GmbH, Langenselbold, Germany) at 1,150 °C according to manufacturer’s instructions. Sulphanilic acid (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany) was used as standard. All analyses were performed in triplicate measurements.
For analyzing elemental composition (C, N, S) samples were dried at 50 °C for 48 h. Afterwards samples were grinded using mortar and pestle. 10 mg of each sample were oxidized in the combustion tube of the elemental analyzer Vario EL II (Elementar Analysensysteme GmbH, Langenselbold, Germany) at 1,150 °C according to manufacturer’s instructions. Sulphanilic acid (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany) was used as standard. All analyses were performed in triplicate measurements.
10
Quantifying Mouse Anti-Rabbit IgG Antibodies
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Serum levels of circulating mouse anti-rabbit IgG were determined by ELISA using mouse IgG quantification sets (Bethyl, Montgomery, Texas, USA). In detail, each well was coated with 250 ng normal rabbit IgG. After blocking, a series of diluted samples was added and incubated for 60 min. Bound Abs were detected by HRP-conjugated goat anti-mouse IgG (Bethyl) and tetramethylbenzidine (Thermo Fisher Scientific). The enzymatic color reaction was stopped by 2 M sulfuric acid (Carl Roth, Karlsruhe, Germany), and the change in OD was measured with a plate reader (PerkinElmer, Waltham, Mass., USA) at 450 nm. Standard reference curves were established using the provided mouse reference sera (Bethyl).
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