C57BL/6J mice were treated intraperitoneally (i.p.) with 90 mg/kg PHZ hydrochloride. After 12 or 30 h of PHZ injection, blood was removed by heart puncture; livers were collected, digested, and macrophages were purified by magnetic-activated cell sorting (MACS) or anti-F4/80 antibody coated Dynabeads. The concentration of heme was quantified in F4/80+ macrophages using the oxalic acid method described by Sassa et al. (27 (link)) using oxalic acid purchased from Sigma-Aldrich (#194131).
Oxalic acid
Manufactured by Merck Group
Sourced in United States, Germany, India, Poland, China, Belgium, United Kingdom, Italy
Oxalic acid is a chemical compound with the formula H2C2O4. It is a colorless crystalline solid that is highly soluble in water. Oxalic acid is commonly used in various industrial and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Citric acid, by Merck Group (49 mentions)
Sodium hydroxide (naoh), by Merck Group (37 mentions)
Acetic acid, by Merck Group (28 mentions)
Malic acid, by Merck Group (26 mentions)
Succinic acid, by Merck Group (23 mentions)
Hydrochloric acid (hcl), by Merck Group (23 mentions)
Methanol, by Merck Group (19 mentions)
Fumaric acid, by Merck Group (18 mentions)
Formic acid, by Merck Group (17 mentions)
Quinic acid, by Merck Group (15 mentions)
Tartaric acid, by Merck Group (14 mentions)
Ascorbic acid, by Merck Group (12 mentions)
Gallic acid, by Merck Group (12 mentions)
Sulfuric acid, by Merck Group (12 mentions)
Lactic acid, by Merck Group (10 mentions)
Ethanol, by Merck Group (10 mentions)
L ascorbic acid, by Merck Group (9 mentions)
Phosphoric acid, by Merck Group (9 mentions)
Acetone, by Merck Group (9 mentions)
Sodium carbonate, by Merck Group (9 mentions)
224 protocols using oxalic acid
1
Quantification of Heme in Macrophages
2
Oxalic Acid Quantification Protocol
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Two standard curves of oxalic acid (99.99% oxalic acid, Sigma-Aldrich Co., St. Louis, MO, USA) were analyzed using the following concentrations: 1, 2, 5, 10, 15 and 25 mg/100 mL, one prepared in 0.2 M HCl and the other prepared in high purity water. The acid standard curve was used for identifying and calculating the total oxalate content, while the water standard curve was used for calculating the soluble oxalate content. All blanks and standard solutions were passed through a 0.45 µm cellulose acetate filter prior to analysis.
3
Oxalic Acid Standard Curve Analysis
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Two standard curves of oxalic acid (99.99% oxalic acid, Sigma‐Aldrich Co., St. Louis, USA) were analyzed, using standards of the following concentrations: 1, 2, 5, 10, 15, and 25 mg/100 ml. One batch of standards was prepared in 0.2 M HCl while the other was prepared in high purity water. The acid standard curve was used for identifying and calculating the total oxalate content, while the water standard curve was used for calculating the soluble oxalate content. All blank and standard solutions were passed through a 0.45‐μm cellulose acetate filter prior to analysis.
4
Isolation and Heme Quantification in Macrophages
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C57Bl/6 mice were treated intraperitoneally (i.p.) with 90 mg/kg phenylhydrazine hydrochloride (PHZ). Livers were collected, and macrophages were purified from cell suspensions by F4/80‐conjugated microbeads and magnetic cell separation columns (Miltenyi) according to the manufacturer's protocol (Miltenyi). The purity of isolated cells was confirmed by FACS and was always >80%. The concentration of heme was quantified in F4/80+ macrophages using the oxalic acid method of Sassa et al. 20 using oxalic acid from Sigma‐Aldrich.
5
Antifungal Activity of WESMS on P. capsici
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P. capsici KACC40180 was obtained from Korea Agricultural Culture Collection in National Institute of Agricultural Sciences, Rural Development Administration, Wanju, Korea. The effect of WESMS on the mycelial growth of P. capsici was determined on potato dextrose agar (PDA) media. PDA powder was added to the WESMS. After autoclaving, 4-day-old mycelial plugs (5 mm diameter) of P. capsici grown on V8 medium were used to inoculate 20 ml of the PDA-WESMS. This was then incubated at 25°C for 7 days. For the control, WENSM replaced the WESMS in the method detailed above. To evaluate the antifungal activity of oxalic acid, final concentration of oxalic acid (Sigma-Aldrich, St. Louis, MO, USA) were adjusted to ensure a range between 100–400 mg/l in PDA. The mycelial plugs (5 mm) were inoculated on the oxalic acid-PDA media, and the mycelial growth rate was checked after 7 days.
6
Platinum Catalyst from Drinking Water Sludge
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The starting material for obtaining the catalysts was obtained from a drinking water treatment plant (ETAP El Atabal, Málaga, Spain) after the flocculation step using a commercial 40% FeCl3 solution. The residue is mainly composed of montmorillonite, iron oxides, humic substances and a minor proportion of Nontronite 5A and calcite. The material was dried at 120 ºC for 12 hours and calcined at 350 and 550 ºC. They were denoted as sludge(120ºC), sludge(350ºC) and sludge(550ºC), respectively.
A platinum/sludge catalyst was prepared by dissolving H2Cl6Pt x H2O (Aldrich 99.9%) in deionised water at 80ºC. The solution was heated to 80ºC and stirred continuously.
An appropriate quantity of oxalic acid (Aldrich 99+%) was added to the solution (oxalic acid/Pt = 3 mol). The ferric sludge calcined at 550ºC was then added to the heated solution and stirred at 80ºC to form a paste. The resulting paste was dried at 110ºC for 16 h. The catalyst contained 1 wt% Pt. The final catalyst was prepared by calcination in static air at 550ºC for 6 h. The catalyst was named as Pt/sludge.
For a comparative purpose a platinum catalyst with the same preparation method and with the same amount of platinum (1 wt.%) was synthesized using aluminium oxide (Sasol. SBET= 179 m 2 g -1 ) as a support. The catalyst was named as Pt/Al2O3.
A platinum/sludge catalyst was prepared by dissolving H2Cl6Pt x H2O (Aldrich 99.9%) in deionised water at 80ºC. The solution was heated to 80ºC and stirred continuously.
An appropriate quantity of oxalic acid (Aldrich 99+%) was added to the solution (oxalic acid/Pt = 3 mol). The ferric sludge calcined at 550ºC was then added to the heated solution and stirred at 80ºC to form a paste. The resulting paste was dried at 110ºC for 16 h. The catalyst contained 1 wt% Pt. The final catalyst was prepared by calcination in static air at 550ºC for 6 h. The catalyst was named as Pt/sludge.
For a comparative purpose a platinum catalyst with the same preparation method and with the same amount of platinum (1 wt.%) was synthesized using aluminium oxide (Sasol. SBET= 179 m 2 g -1 ) as a support. The catalyst was named as Pt/Al2O3.
7
HPLC Analysis of Organic Acids
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A high-performance liquid chromatography (HPLC) system (LC-10A, Shimadzu, Tokyo, Japan) and Capcell Pak C18 column (4.6 mm × 250 mm, 5 μm; Shiseido Co., Tokyo, Japan) were used. A high-speed refrigerated centrifuge was used (KDC-140HR, Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd., Hefei, China), along with a constant-temperature culture oscillator (ZWY-211B, Shanghai Zhicheng Analytical Instrument Manufacturing Co., Ltd., Shanghai, China).
Oxalic acid, tartaric acid, formic acid, malic acid, lactic acid, acetic acid, citric acid, and succinic acid standards were obtained from Sigma (chromatographically pure; Sigma-Aldrich, St. Louis, MO, USA). Methanol (chromatographically pure, Tianjin Science and Europe Chemical Reagent Co., Ltd., Tianjin, China) and phosphoric acid (chromatographically pure, Chengdu Jinshan Chemical Reagent Co., Ltd., Chengdu, China) were obtained from commercial sources.
Oxalic acid, tartaric acid, formic acid, malic acid, lactic acid, acetic acid, citric acid, and succinic acid standards were obtained from Sigma (chromatographically pure; Sigma-Aldrich, St. Louis, MO, USA). Methanol (chromatographically pure, Tianjin Science and Europe Chemical Reagent Co., Ltd., Tianjin, China) and phosphoric acid (chromatographically pure, Chengdu Jinshan Chemical Reagent Co., Ltd., Chengdu, China) were obtained from commercial sources.
8
Antioxidant and Phytochemical Evaluation
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All reagents and chemicals including gallic acid, rutin, quercetin, aluminum chloride (AlCl3), ferric chloride (FeCl3·7H2O), Folin-Ciocalteu, oxalic acid, trichloroacetic acid (TCA), sulfuric acid (H2SO4), hydrochloric acid (HCl), Na2CO3, ammonium molybdate, potassium ferricyanide (K3Fe(CN)6), catechin, 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2′-azinobis (3-ethylobenzothiazoline-6-sulphonic acid diammonium salt) (ABTS) are products of Sigma-Aldrich, South Africa. Others such as anhydrous sodium carbonate, sodium nitrite (NaNO2), ethanol, methanol, n-butanol, sodium acetate, butylated hydroxyoluene (BHT), diethyl ether, glacial acetic acid, sulfanilic acid, potassium persulfate, and sodium nitroprusside were also purchased from Sigma-Aldrich. All the reagents used were of analytical grade.
9
Extraction and Analysis of Bioactive Compounds from Glycyrrhiza Japonica
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Whole plants of GJ were purchased from Tongling Hetian Chinese Medicine Pieces Co. Ltd. (Anhui, China) and smashed into powder with a disintegrator (Tianjin Tester Instrument Co, Ltd., Tianjin, China). Analytical grade reagents including choline chloride (≥98% purity), betaine (≥98% purity), oxalic acid (≥98% purity), DL-malic acid (≥98% purity), citric acid (≥98% purity), and malonic acid (≥98% purity) were purchased from Sigma-Aldrich (Shanghai, China). Analytical pure methanol, ethanol, and acetone were purchased from Tianjin Tianli Chemical Reagent Co. Ltd. (Tianjin, China). EA (≥98% purity) was purchased from Sichuan Vicky Biotechnology Co., Ltd. (Sichuan, China). DPPH (≥98% purity) was purchased from Diyibio Biotechnology Co., Ltd. (Shanghai, China). The chromatographic grade methanol was purchased from Honeywell (Arizona, USA). The chromatographic grade phosphoric acid (≥98% purity) was purchased from Tianjin Kemiou Chemical Reagent Co., Ltd (Tianjin, China). Ultra-pure water produced by Milli-Q IQ7000 (Millipore Corp, USA) was used throughout the experiment.
10
Quantitative Analysis of Organic Acids
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In the present study, chemicals with analytical purity were used. Organic acid standards (citric acid, tartaric acid, oxalic acid, malic acid, succinic acid and fumaric acid), sugar standards (glucose, fructose, and sucrose), and vitamin C standards (l-ascorbic acid) were obtained from Sigma–Aldrich (St. Louis, MO, USA). The other chemicals were obtained from Merck (Darmstadt, Germany).
For organic acid extraction, the method by Bevilacqua and Califano [18 (link)] was modified. About 200 g of samples was fragmented and 10 g from each sample was delivered to centrifuge tubes. The 10 ml of 0.009 N H2SO4 was added to the samples and the samples were homogenized with Heidolph Silent Crusher M, Germany. Then, the samples were mixed for an hour with a shaker (Heidolph Unimax 1010, Germany) and centrifuged at 14.000 × rpm for 15 min. The supernatants were passed through coarse filter paper, then twice through a 0.45 mm membrane filter (Millipore Millex-HV Hydrophilic PVDF, Millipore, USA), and last in a SEP-PAK C18 cartridge. The concentration of organic acids was determined by HPLC using an Aminex column (HPX-87H, 300 mm × 7.8 mm, Bio-Rad) fitted on an Agilent 1100 series HPLC G 1322 A, Germany) [18 (link)]. Organic acids were detected at 214 and 280 nm wavelengths. The mobile phase, 0.009 N H2SO4 was passed through a 0.45 μm filter membrane.
For organic acid extraction, the method by Bevilacqua and Califano [18 (link)] was modified. About 200 g of samples was fragmented and 10 g from each sample was delivered to centrifuge tubes. The 10 ml of 0.009 N H2SO4 was added to the samples and the samples were homogenized with Heidolph Silent Crusher M, Germany. Then, the samples were mixed for an hour with a shaker (Heidolph Unimax 1010, Germany) and centrifuged at 14.000 × rpm for 15 min. The supernatants were passed through coarse filter paper, then twice through a 0.45 mm membrane filter (Millipore Millex-HV Hydrophilic PVDF, Millipore, USA), and last in a SEP-PAK C18 cartridge. The concentration of organic acids was determined by HPLC using an Aminex column (HPX-87H, 300 mm × 7.8 mm, Bio-Rad) fitted on an Agilent 1100 series HPLC G 1322 A, Germany) [18 (link)]. Organic acids were detected at 214 and 280 nm wavelengths. The mobile phase, 0.009 N H2SO4 was passed through a 0.45 μm filter membrane.
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