This assay was performed as described before52 (link),53 (link). Briefly, a 0.2 M NaOH with 40% pyridine solution was made fresh using a 1 M NaOH stock and 100% pyridine solution (Sigma–Aldrich, St. Louis, MO). 5 µL (i.e., 1/200) of 0.1 M potassium ferricyanide stock solution was added to 495 µL of the aforementioned NaOH + pyridine mix to generate the pyridine hemochrome assay solution. 50 µL of the assay solution was mixed with 50 µL of TBS buffer (50 mM Tris-HCl, 150 mM NaCl, pH = 7.4) and used as a blank. Next, 50 µL of the assay solution was mixed with 50 µL of MmcA in TBS buffer, and UV-vis scans were immediately performed using a Shimadzu 1900i (Shimadzu, Torrance, CA, USA) to record the oxidized spectra. A 10 mM stock solution of sodium dithionate was added to the protein assay mixture and UV-vis scans were performed using a Shimadzu 1900i (Shimadzu, Torrance, CA, USA) to record the fully reduced pyridine hemochrome spectra.
Pyridine solution
Manufactured by Merck Group
Sourced in Macao
Pyridine solution is a laboratory reagent used for various chemical applications. It is a clear, colorless liquid that serves as a solvent, reagent, and intermediate in organic synthesis. Pyridine solution is widely used in the pharmaceutical, agrochemical, and chemical industries for tasks such as extraction, purification, and derivatization of organic compounds.
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Lab products found in correlation
N methyl n trimethylsilyl trifluoroacetamide mstfa, by Merck Group (1 mentions)
Tetrahydrofuran solution, by Merck Group (1 mentions)
N heptane hplc grade, by Merck Group (1 mentions)
α tocopherol, by Merck Group (1 mentions)
Sodium methoxide solution, by Merck Group (1 mentions)
N methyl n trimethylsilyltrifluoroacetamide mstfa, by Thermo Fisher Scientific (1 mentions)
Hplc grade, by Merck Group (1 mentions)
N hexane, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Sodium acetate, by Avantor (1 mentions)
6 protocols using pyridine solution
1
Pyridine Hemochrome Assay for MmcA
2
Pyridine Hemochrome Assay for Protein
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This assay was performed as described before (56 (link), 57 ). Briefly, a 0.2 M NaOH with 40% pyridine solution was made fresh using a 1 M NaOH stock and 100% pyridine solution (Sigma Aldrich, St. Louis, MO). 5 µL (i.e., 1/200) of 0.1 M potassium ferricyanide stock solution was added 495 µL of the aforementioned NaOH + pyridine mix to generate the pyridine hemochrome assay solution. 50 µL of the assay solution was mixed with 50 µL of TBS buffer (50 mM Tris-HCl, 150 mM NaCl, pH = 7.4) and used as a blank. Next, 50 µL of the assay solution was mixed with 50 µL of MmcA in TBS buffer, and UV-vis scans were immediately performed using a Shimadzu 1900i (Shimadzu, Torrance, CA, USA) to record the oxidized spectra. A 10 mM stock solution of sodium dithionate was added to the protein assay mixture and UV-vis scans were performed using a Shimadzu 1900i (Shimadzu, Torrance, CA, USA) to record the fully reduced pyridine hemochrome spectra.
3
Metabonomics Analysis of Mesencephalic Tissue and Serum
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Derivatization was performed by adding 25 μL and 100 μL, respectively, for mesencephalic tissue and serum sample, of methoxyamine hydrochloride in pyridine solution (10 mg/mL) (Sigma-Aldrich, St. Louis, MO, USA) to dried brain samples at 70 °C. After 1 h, 50 and 100 μL, respectively, for mesencephalic tissue and serum sample of N-Methyl-N-(trimethylsilyl)-trifluoroacetamide (MSTFA, Sigma-Aldrich, St. Louis, MO, USA) were added and the samples were left at room temperature for 1 h. Samples were then diluted in hexane (50 μL for the mesencephalic tissue and 600 μL for the serum sample) with an internal standard (undecane at 25 ppm). For the serum samples, diluted samples were then filtered (PTFE 0.45 μm) and transferred into glass vials. As before, sample blanks were made to avoid noise caused by the laboratory instruments or by the chemicals used for the preparation, by following the same procedure.
4
Characterization of Malaysian Palm Oil
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Fresh commercial crude PMFO was obtained from a local palm oil mill in Malaysia. Sodium hydroxide (NaOH) , n-hexane, and n-heptane (HPLC grade) were purchased from Merck KGaA, Germany while 85％ phosphoric acid, ammonium monovanadate and ammonium heptamolybdate tetrahydrate were bought from Friendemann Schmidt Chemical, Germany. Natural bleaching earth (NBE) was purchased from Taiko Bleaching Earth Sdn. Bhd., Malaysia, Magnesium oxide, potassium dihydrogen phosphate, and 6N of nitric acid were acquired from Chemiz (M) Sdn. Bhd., Malaysia, Potassium hydrogen phthalate was bought from Classic Chemicals Sdn. Bhd., Malaysia. Isopropanol was purchased from Qrec (Asia) Sdn. Bhd., Malaysia, Sodium methoxide solution, tetrahydrofuran solution, and pyridine solution (HPLC grade) were purchased from Sigma-Aldrich Corporation, Germany. n-heptane (GC grade) and N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA) were obtained from Acros Organics, Belgium. 1-glyceryl monononadecanoin, 1,3-glyceryl dinonadecanoin and glyceryl trinonadecanoin powders were bought from Nu-Chek Prep, Inc., USA. Analytical grade α-tocopherol (α-T, 95％) used was from Sigma-Aldrich Corporation, Germany and α-tocotrienol (α-T3, 99.1％) , β-tocotrienol (β-T3, 99.9％) , γ-tocotrienol (γ-T3, 98.3％) , and δ-tocotrienol (δ-T3, 95.5％) were from Chromadex, USA.
5
Heme Biosynthesis Assay Protocol
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Pyridine solution (Sigma), hydroxyethylpiperazine-N-[2-ethanesulphonic acid] (HEPES) solution 20 mM, 0.1M NaOH, 1M HCl, chloroquine phosphate, bovine heme [Fe(III)PPX], glacial acetic acid, sodium acetate (BDH Chemicals, England), methanol, saturated acetate solution (pH5.0).
6
Characterizing Silica Shell Integrity
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The presence of any pinholes in the SHINs' silica shells was investigated by running electrochemical and Raman tests. Cyclic voltammetry in 0.1 M H 2 SO 4 was performed on a glassy carbon electrode with SHINs drop casted on the surface, sweeping the potentials between À0.2 V and 1.5 V (vs. Ag/AgCl) using a Biologic potentiostat. 10 For the Raman pinhole test, the nanoparticle solution was deposited on a silicon wafer (Si (100), Agar Scientic) using 10 mM pyridine solution (Sigma Aldrich) as a probe molecule. 17, 18 The spectra were collected with a Renishaw inVia microscope using a laser with a 632.8 nm wavelength.
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