Dilinoleoylphosphatidylcholine (DLPC), dimyristoylphosphatidylcholine (DMPC), 1-palmitoyl-2-arachidonoylphosphochatidyline (PAPC), 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), and 1-palmitoyl-2-(16-doxylstearoyl)phosphatidylcholine (16-PC) were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL). Zea was purchased from CaroteNature (Lupsingen, Switzerland), malondialdehyde (MDA) was obtained from Cayman Chemical (Ann Arbor, MI). CTPO (3-Carbamoyl-2,2,5,5-tetramethyl-3-pyrrolin-1-oxyl), TEMPO (2,2,6,6-Tetramethylpiperidine 1-oxyl), TEMPONE (2,2,6,6-tetramethylpiperidone-N-oxyl), and other chemicals, of at least reagent grade, were purchased from Sigma-Aldrich Co. (St. Louis, MO).
Malondialdehyde mda
Manufactured by Cayman Chemical
Sourced in United States
Malondialdehyde (MDA) is a chemical compound used in analytical and research applications. It serves as a marker for oxidative stress and lipid peroxidation. MDA can be quantified using various analytical techniques, including spectrophotometry and chromatography.
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Lab products found in correlation
Tempo, by Merck Group (1 mentions)
Tempone, by Merck Group (1 mentions)
Dimyristoylphosphatidylcholine, by Avanti Polar Lipids (1 mentions)
1 palmitoyl 2 oleoylphosphatidylcholine, by Avanti Polar Lipids (1 mentions)
1 palmitoyl 2 16 doxyl stearoyl phosphatidylcholine 16 pc, by Avanti Polar Lipids (1 mentions)
Microtube homogenizer, by Benchmark Scientific (1 mentions)
8 iso prostaglandin f2α, by Cayman Chemical (1 mentions)
L ascorbic acid, by Merck Group (1 mentions)
Protein carbonyl colorimetric assay, by Cayman Chemical (1 mentions)
Serum bovine albumin, by Merck Group (1 mentions)
Z36hk, by Hermle (1 mentions)
Bradford colorimetric assay, by Merck Group (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Superoxide dismutase, by Cayman Chemical (1 mentions)
Bca assay kit, by Thermo Fisher Scientific (1 mentions)
8 protocols using malondialdehyde mda
1
Lipid Synthesis and Antioxidant Analysis
2
Assessing Wound Antioxidant and Lipid Peroxidation
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Wound tissue samples were excised at the size of 1.0 cm × 1.0 cm on day 2 and day 8 post-wounding and biochemically analyzed to estimate endogenous antioxidant enzyme and lipid peroxidation activities. Each tissue sample was dried and weighed prior to use for analysis. Tissues were transferred to a bead tube with Tris-buffer and homogenized using a microtube homogenizer (Benchmark Scientific, USA). The homogenized tissues were then centrifuged at 6000 rpm for 20 min and we collected the supernatant for the biochemical estimations. Activity levels of endogenous antioxidant enzymes such as superoxide dismutase (SOD) (Item No 706002; Cayman) and glutathione peroxidase (GPx) (Item No 703102; Cayman) and lipid peroxidation, measured as malondialdehyde (MDA) content (Item No 10009055; Cayman), were measured following the vendor’s protocol.
3
Evaluation of Plasma Antioxidant-Oxidant Status
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Plasma antioxidant-oxidant (redox) status was evaluated by measuring total antioxidant capacity (TAC), lipid peroxidation (TBARS), extracellular SOD (EC-SOD) activity, nitrites and nitrates (NOX) levels, glutathione (GSH) and oxidized glutathione (GSSG) as previously described [19 (link),20 (link)]. To avoid technical variations samples from non-exposed and exposed workers were always included for each determination. Specifically, TAC was measured by the ABTS ((2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) method with a commercial kit (Cayman Chemical, Ann Arbor, MI, USA). Lipid peroxidation was evaluated by measuring the formation of thiobarbituric acid reactive substances (TBARS, including malondialdehyde (MDA) which are formed as a byproduct of lipid peroxidation (Cayman Chemical, Ann Arbor, MI, USA). Determination of EC-SOD activity was based on the dismutation of superoxide oxygen and hydrogen peroxide with a commercial kit (Cayman Chemical, Ann Arbor, MI, USA) Nitrites (stable end-product of NO) and nitrate (NOX) levels were measured by nitrate reductase and Griess reaction [21 (link)]. GSH and GSSG were measured by the liquid chromatography tandem mass spectrometry (LC-MS/MS) system [20 (link)].
4
Antioxidant and Lipid Peroxidation Assays
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Drugs and reagents were obtained from the following sources: 8-iso prostaglandin-d4 F2α-d4 (8-isoPGF2α–d4), 8-iso prostaglandin F2α (8-isoPGF2α - F2-isoprostanes), 4-hyroxynonenal (4-HNE), 4-hydroxyhexenal (4-HHE), 4-oxo-nonenal (4-ONE), and malondialdehyde (MDA) from Cayman Chemical Company (Ann Arbor, MI, USA), cis-4,7,10,13,16,19-docosahexaenoic acid (DHA), glutathione peroxidase (GSH-Px), l -glutathione reduced (GSH), (±) α-tocopherol (vitamin E), all trans retinol (vitamin A), and L-ascorbic acid (vitamin C) from Merck KGaA (Sigma–Aldrich, Darmstadt, Germany).
5
Oxidative Stress Biomarkers Assessment
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Oxidants can react with endogenous proteins, lipids, and other materials to form oxidation products. Protein carbonylation was detected using a protein carbonyl colorimetric assay (Cayman Chemical, Ann Arbor, MI) and measured at 370 nm. The detection of lipid peroxides in the form of malondialdehyde (MDA; Cayman Chemical, Ann Arbor, MI) measured at 535 nm, and 4-Hydroxyl-2-noneal (4-HNE) and neuron specific enolase (NSE) (MyBioSource, Inc., San Diego, CA) measured at 540 nm and 450 nm, respectively. Protein nitrotyrosine concentration (StressMarq Biosciences Inc., Victoria, BC, Canada) was measured with 1:2 plasma dilution and read at 450 nm. The assays were conducted according to manufacturer’s instructions without other modifications not mentioned.
6
Biochemical Assays for Muscle Tissue
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For biochemical assays, muscle tissue (100 mg/mL) was homogenized in RIPA buffer (Sigma-Aldrich, Saint Louis, MO, USA) and centrifuged at 1600× g for 10 min at 4 °C (Z 36 HK, Hermle-Labortechnik, Wehingen, Germany). Supernate was collected for determining the thiobarbituric-acid-reactive substances (TBARS), total peroxidase, and glutathione peroxidase activities.
TBARS levels were calculated based on the standard curve of malondialdehyde (MDA) (Cayman Chemical, Ann Arbor, MI, USA) (0 to 50 µM) and followed the protocol described previously [11 (link)]. TBARS levels (μM) in muscle tissue were normalized by protein concentration (mg/mL) and expressed as μM/mg protein.
Total peroxidase activity was measured by the disappearance of hydrogen peroxide (H2O2) at 240 nm [12 (link)], whilst the total glutathione peroxidase (GPx) activity was determined according to previously standardized methods [13 (link)]. Total peroxidase and GPx activities were normalized by protein concentration (mg/mL) and expressed in mU/mg protein.
Total protein concentration of the muscle homogenates was determined by Bradford colorimetric assay at 595 nm (Sigma-Aldrich, Saint Louis, MO, USA) employing serum bovine albumin (Sigma-Aldrich, Saint Louis, MO, USA) (0 to 1.4 mg/mL) as standard [14 (link)].
TBARS levels were calculated based on the standard curve of malondialdehyde (MDA) (Cayman Chemical, Ann Arbor, MI, USA) (0 to 50 µM) and followed the protocol described previously [11 (link)]. TBARS levels (μM) in muscle tissue were normalized by protein concentration (mg/mL) and expressed as μM/mg protein.
Total peroxidase activity was measured by the disappearance of hydrogen peroxide (H2O2) at 240 nm [12 (link)], whilst the total glutathione peroxidase (GPx) activity was determined according to previously standardized methods [13 (link)]. Total peroxidase and GPx activities were normalized by protein concentration (mg/mL) and expressed in mU/mg protein.
Total protein concentration of the muscle homogenates was determined by Bradford colorimetric assay at 595 nm (Sigma-Aldrich, Saint Louis, MO, USA) employing serum bovine albumin (Sigma-Aldrich, Saint Louis, MO, USA) (0 to 1.4 mg/mL) as standard [14 (link)].
7
Liver Biomarkers Assessment
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The following biomarkers in liver tissue were determined using commercial kits according to manufacturer's instructions. Malondialdehyde (MDA; catalog no. 10009055; Cayman Chemical, Ann Arbor, MI) and ROM (catalog no. STA-347; Cell Biolabs, San Diego, CA) were assessed through a fluorometric methods; GSH through a kinetic colorimetric assay (catalog no. NWK-GSH01; Northwest Life Science Specialties, Vancouver, WA); and SOD through a spectrophotometric method (catalog no. 706002; Cayman Chemical).
8
Oxidative Stress Markers in Adipose Tissue
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Superoxide dismutase (catalog no. 706002; Cayman Chemical, Ann Arbor, MI), malondialdehyde (MDA; catalog no. 10009055; Cayman Chemical), GSH (catalog no. NWK-GSH01; Northwest Life Science Specialties, Vancouver, WA), and ROS (catalog no. STA-347, Cell Biolabs, San Diego, CA) were analyzed in subcutaneous adipose tissue using commercial kits. Concentration of total protein of the adipose samples was measured using the BCA assay kit (catalog no. 23227; Thermo Fisher Scientific).
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