Sodium pyruvate, preparation

Diethyl oxalate (29.2 g, 0.20mol) and 4-bromo-2-nitrotoluene (21.6 g, O.lOmol) were added to a cooled solution of sodium cthoxide prepared from sodium (4.6 g, 0.20 mol) and ethanol (90 ml). The mixture was stirred overnight and then refluxed for 10 min. Water (30 ml) was added and the solution refluxed for 2h to effect hydrolysis of the pyruvate ester. The solution was cooled and concentrated in vacuo. The precipitate was washed with ether and dried. The salt was dissolved in water (300 ml) and acidified with cone. HCl. The precipitate was collected, washed with water, dried and recrystallizcd from hexane-EtOAc to give 15.2 g of product. 

A) Ethyl Butyryl-Pyruvate 146 grams of ethyl oxalate are condensed with 86 grams of methyl-(n)-propyl-ketOne in the presence of sodium ethylate prepared from 25 grams of sodium. 135 grams of product, having a boiling point of 1l3°C/6 mm, are obtained. 

Media basal RPMI 1640 medium requires supplementation with sodium pyruvate, L-glutamine, and penicillin/streptomycin before use. The supplements are supplied as concentrates of 50X or 100X, and the appropriate amounts should be added. Standard tissue culture media for hybridoma production contain 5%, 10%, or 15% fetal bovine serum (FBS see Note 6). Sufficient quantities should be prepared in advance and a sterility check should be performed on them prior to use. 

Reading (1982) adapted this to obtain concentrates of NSF for in vitro immunization. For this purpose, thymocytes are prepared (Section 5.4.2.4) from both BALB/c and C57BL/6 mice and equal numbers are mixed to a final concentration of 5x 10 cells/ml and cultured in plastic culture flasks in DMEM supplemented with 4.5 g dextrose/1, MEM non-essential amino acids, sodium pyruvate, 2-ME, HAT, HEPES, and 2% rabbit serum. After 48 h, the cells and other debris are pelleted and the supernatant (thymocyte-conditioned medium, TCM) is filtered (200 nm) and stored at — 70°C in aliquots of 10 ml. 

O-Prolected sugar thioformamides have also been prepared in 44-87% yields by tri-n-butyltin hydride reduction of isothiocyanate precursors in ether.156 For the preparation of 5-A-thioacylneuraminic acids (84), a chemoenzymatic route based on the M-acetylneuraminate pyruvate lyase-mediated condensation of the corresponding A -thioacyl-n-mannosamine derivatives (83) and sodium pyruvate has been reported.189 The enzyme-catalyzed aldol reaction was performed at pH 6.8 and afforded the desired compound in 55% yields (Scheme 24). 

Pure cultured astrocytes were prepared by a modification of the method of Booher and Sensenbrenner (32). Cerebral hemispheres from newborn Wistar rats were mechanically dissociated in Waymouth s medium supplemented with sodium pyruvate (110 mg/ml), antibiotics and 10 % heat-inactivated fetal calf serum. Cells from one brain were seeded in 6 Petri dishes (100 mm diameter) and the cultures were maintained in the serum containing medium for 21 days and then switched for 3 days either to a medium containing thyroid hormone depleted serum (33) or to a serum-free chemically defined medium. This consisted of Waymouth s medium supplemented with 5 pg/ml insulin and 0.5 Mg/ml fatty acid-free bovine serum albumin and antibiotics. Pure cultured astrocytes were also prepared from 14-day-old embryonic chick brain as described above (32). 

To achieve kinetic resolution of mexiletine (di-o-methyl-phenoxyisopropylamine), lyoph-ilized E.coli cells containing Chromobacterium violaceum (S)-selective o>TA (100 mg, prepared as described in Ref. [75]) are resuspended in phosphate buffer (20 mL, 100 mM, 0.9 mM sodium pyruvate, 1 mM PLP). Crotalus adamanteus i-amino acid oxidase (30 mg, 9 U, Sigma) is rehydrated in the buffer. (R/S)-mexiletine (100 mg, 23 mM)... 

Methylsuccinic acid has been prepared by the pyrolysis of tartaric acid from 1,2-dibromopropane or allyl halides by the action of potassium cyanide followed by hydrolysis by reduction of itaconic, citraconic, and mesaconic acids by hydrolysis of ketovalerolactonecarboxylic acid by decarboxylation of 1,1,2-propane tricarboxylic acid by oxidation of /3-methylcyclo-hexanone by fusion of gamboge with alkali by hydrog. nation and condensation of sodium lactate over nickel oxide from acetoacetic ester by successive alkylation with a methyl halide and a monohaloacetic ester by hydrolysis of oi-methyl-o -oxalosuccinic ester or a-methyl-a -acetosuccinic ester by action of hot, concentrated potassium hydroxide upon methyl-succinaldehyde dioxime from the ammonium salt of a-methyl-butyric acid by oxidation with. hydrogen peroxide from /9-methyllevulinic acid by oxidation with dilute nitric acid or hypobromite from /J-methyladipic acid and from the decomposition products of glyceric acid and pyruvic acid. The method described above is a modification of that of Higginbotham and Lapworth. ... 

Methylindole has been prepared from the a5-methylphenyl-hydrazone of pyruvic acid, by the action of sodium amide or sodium hydride on indole followed by methyl iodide at elevated temperatures,by treatment of indole with methyl p-toluene-sulfonatc and anhydrous sodium carbonate in boiling xylene, and by the action of inelhyl sulfate on indole previously treated... 

The sodium or potassium salt of 6-azauracil in aqueous ethanol, anhydrous ethanol, or ethylene glycol reacted with methyl iodide practically exclusively to give the 3-methyl derivative (63). In toluene the sodium, potassium, and mercuric salts produced no methylated derivatives whereas the silver salt also yielded the 3-methyl derivative, Similarly, the 3-methyl derivative was prepared from the mercuric salt of 6-azathymine, and its structure was established by hydrolysis to pyruvic acid 4-methylthiosemicarbazone. ... 

Nef prepared acetol in several ways, the more important of which depended upon the reaction between bromoacetone and potassium or sodium formate or acetate, and the subsequent hydrolysis of the ester by methyl alcohol.1 2 Acetol is also formed, together with pyruvic acid, by the direct oxidation of acetone by Baeyer and Villiger s acetone-peroxide reagent.3... 

Methylquinoxaline may be prepared in 88-92% yields from o-phenylenediamine and pyruvic aldehyde-sodium bisulfite by this same procedure. 

Semicarbazones are prepared by dissolving semicarbazide hydrochloride (ca Ig) and sodium acetate (ca 1.5g) in water (8-lOml) and adding the aldehyde or ketone (0.5-lg) and stirring. The semicarbazone crystallises out and is recrystallised from ethanol or aqueous ethanol. These are hydrolysed by steam distillation in the presence of oxalic acid or better by exchange with pyruvic acid (Hershberg dOC 13 542 1948). 

Serum Preparation Whole blood was collected after decapitation of the animal and allowed to clot at room temperature for 30 min. The serum was then separated from the clot by centrifugation at 5000 rpm at U C. Aliquots of the serum were immediately taken and refrigerated for the determination of glutamic-pyruvic transaminase and lactic dehydrogenase. An additional 0.5 ml of serum from each animal was treated with a few milligrams of sodium fluoride and refrigerated for later glucose determinations. The rest of the serum was frozen for the remainder of the clinical chemistry determinations. 

A solution of Grignard reactive (prepared from 247 g of o-bromoanisole and 31.6 g of magnesium in 1500 ml of ether) is added dropwise to a solution of 91 g of diphenyl pyruvic acid in ether. The mixture is refluxed for 2 hours. After cooling to the reaction mixture is added 5% hydrochloric acid and the product is extracted with 5% solution of sodium hydroxide. a-(o-Anisyl)-p,15-diphenyl lactic acid is precipitated by addition of concentrated hydrochloric acid yield 62 g, melting point 194-195°C. 

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