Diazomethane was prepared by treating N-methyl-N-nitroso-p-toluenesulfonamide (Diazald®, Sigma-Aldrich, St Louis, MO, USA) with sodium hydroxide (Vetec, Xerém, Brazil) in ethyl ether (Merck, Darmstadt, GE). Dichloromethane was purchased from Tedia (Fairfield, OH, USA). Ethanol p.a. absolute was obtained from Merck. UA (>90% purity, article U6753), OA (>97% purity, article O5504) and BA (90% purity, article 91466) were purchased from Sigma-Aldrich Co to be used as standards.
Diazomethane
Manufactured by Merck Group
Sourced in United States, Germany
Diazomethane is a colorless, highly reactive gas that is commonly used as a methylating agent in organic synthesis. It is a versatile laboratory reagent that can be used to introduce a methyl group (-CH3) into a wide range of organic compounds. Diazomethane is a powerful and potentially hazardous substance, and its use requires appropriate safety precautions and handling procedures.
Automatically generated - may contain errors
2 protocols using diazomethane
1
Diazomethane Synthesis Protocol
2
Quantification of Free Fatty Acids in Milk
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The presence of free fatty acids (FFAs) in milk samples was determined following plasma application. Lipid extraction was carried out according to the method described by Lopez-Lopez, Castellote-Bargallo, and Lopez-Sabater (2001) with some modifications and standards used as previously described by (Lanciotti et al., 2006) . Dichloromethane-methanol (2:1) (Sigma, Munich, Germany) was added to milk and the mixture was mechanically agitated in a shaker (Hotech, Taipei, Taiwan) and then centrifuged at 3000 × g. After washing with sterile distilled water, the organic phase was filtered and the solvent was removed in a rotatory evaporator (Heidolph, Schwabach, Germany).
Lipids were extracted by methylation using n-hexane (Merck, Darmstadt, Germany) and 2 M potassium hydroxide in methanol (Merck, Darmstadt, Germany). After evaporation under N 2 flux, diazomethane (Sigma, Munich, Germany) was added. Fatty acid methyl ester analysis was carried and Wiley version 6 Mass spectral data base. The quantification of FFA level of milk samples was performed using C11:0 as an internal standard at concentration of 100 µL per 2.5 mg of fatty acid esters.
Lipids were extracted by methylation using n-hexane (Merck, Darmstadt, Germany) and 2 M potassium hydroxide in methanol (Merck, Darmstadt, Germany). After evaporation under N 2 flux, diazomethane (Sigma, Munich, Germany) was added. Fatty acid methyl ester analysis was carried and Wiley version 6 Mass spectral data base. The quantification of FFA level of milk samples was performed using C11:0 as an internal standard at concentration of 100 µL per 2.5 mg of fatty acid esters.
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