Extraction thiamin

Stomach extract Streptococcus lactis extract Thiamine HCI Thiamine nitrate Thymus extract... 

L-Alanine Carotene Cholecalciferol Choline bitartrate Chondroitin sulfate Copper iodide (ous) DL-Leucine Linolenic acid Olive (Olea europaea) oil Pancreatin D-Phenylalanine DL-Phenylalanine Propionic anhydride Silicon Sodium chloride Sugar cane (Saccharum officinarum) extract Thiamine HCI Vipers bugloss (Echium plantagineum) oil... 

To a solution of 20 parts of thiamine hydrochloride in 30 parts of water is added an aqueous solution of sodium hydroxide (7.2 parts of NaOH in 30 parts of water), and the mixture is cooled with water. The mixture is allowed to stand for 30 minutes, 60 parts of chloroform is added, followed by a solution of 30 parts of crude sodium tetrahydrofurfurylthiosulfate in 30 parts of water, and the whole is stirred for 30 minutes. The chloroform layer is separated and the aqueous layer is extracted twice with 20 parts of chloroform. All the chloroform solutions are combined and shaken with 50 parts of 5% hydrochloric acid. The acid solution is decolorized and neutralized with alkali carbonate, whereupon thiamine tetrahydrofurfuryl disulfide separates out in the resinous state but soon solidifies [MP 129 (decomp.)]. The yield is 16 parts. Recrystallization from ethyl acetate gives colorless prisms melting at 132°C (decomp.). 

An extract from the soluble stromal proteins of purified and intact spinach-leaf chloroplasts was prepared by lysis of the cells in buffer, centrifugation of the suspension of broken cells, and concentration of the supernatant with removal of insoluble material. This extract contained all of the enzymes involved in the condensation of the cyclic moieties of thiamine, thiazole, and pyramine. Thus, the synthesis of thiamine in this extract following the addition of pyramine and putative precursors was a proof that the system had the possibility of building the thiazole. It was found that L-tyrosine was the donor of the C-2 carbon atom of thiazole, as in E. coli. Also, as in E. coli cells, addition of 1 -deoxy-D-f/irco-pen-tulose permitted synthesis of the thiamine structure. The relevant enzymes were localized by gel filtration in a fraction covering the 50- to 350-kDa molecular-mass range. This fraction was able to catalyze the formation of the thiazole moiety of thiamine from 0.1 -mM 1-deoxy-D-t/ireo-pentulose at the rate of 220 pmol per mg of protein per hour, in the presence of ATP and Mg2+. 

The significance of these metabolites in the biosynthesis of the thiamine thiazole in considered next. Although, from their constitution, and from the tracer experiments, the metabolites are undoubtedly the products of transformation of 1-deoxy-D-t/ireo-pentulose, their significance in the biosynthesis of the thiazole of thiamine is not clear. The thiazole glycol is not a product arising from a transformation of the thiazole (5) of thiamine. Reduction to this thiazole (5) occurs in dialyzed extracts of disrupted cells, in the presence of ATP, NADH, and NADPH, but only at 0.2% the rate of synthesis of the thiamine thiazole (5) by intact cells. The behavior of the thiazole glycol on plates is merely a consequence of the extreme sensitivity of the tetrazolium reagent. 

Thiamine was biosynthesized by resting cells of S. typhimurium strain thilO/T-ath-383, which can synthesize thiamine from exogenous glucose, AIRs, and thiazole.54 Derepression was achieved by conventional means. The organism was cultivated in the presence of a suboptimal amount of thiamine (20 nM), the washed cells were resuspended in a minimal medium containing glucose (10 mM), thiazole (1-2 mM), and labeled AIRs (10 p,M). During the incubation (1.5 hours 37°C), the level of thiamine diphosphate in the cells had risen from about 0.04 to 0.5 nmol/mg. In work with molecules labeled with stable isotopes, thiamine was extracted and cleaved by ethanethiol to 4-amino-5-(ethyl-... 

Vitamins such as thiamin, biotin, and vitamin Bj2 are often added. Once again, the requirements of anaerobes are somewhat greater, and a more extensive range of vitamins that includes pantothenate, folate, and nicotinate is generally employed. In some cases, additions of low concentrations of peptones, yeast extract, casamino acids or rumen fluid may be used, though in higher concentrations, metabolic ambiguities may be introduced since these compounds may serve as additional carbon sources. 

The SP procedure of water-soluble vitamins from multivitamin tablets is particularly challenging due to the diverse analytes of varied hydrophobicities and pfC. Water-soluble vitamins (WSVs) include ascorbic acid (vitamin C), niacin, niacinamide, pyridoxine (vitamin B ), thiamine (vitamin Bj), folic acid, riboflavin (vitamin B2) and others. While most WSVs are highly water soluble, riboflavin is quite hydrophobic and insoluble in water. Folic acid is acidic while pyridoxine and thiamine are basic. In addition, ascorbic acid is light sensitive and easily oxidized. The extraction strategy employed was a two-step approach using mixed solvents of different polarity and acidity as follows ... 

Typical FI A manifolds are shown in Fig. 13.10 with two general alternatives depending on whether injection takes place before or after the continuous extractor device. The most common situation is when prior injection of the sample take place. Figure 13.10(a) depicts a manifold for the determination of vitamin Bi in pharmaceuticals [178], based on the oxidation of thiamine to thiochrome in a carrier of potassium ferricyanide in a basic medium (NaOH). The thiochrome is continuously extracted into a chloroform stream and the fluorescence of the organic phase is measured continuously. 

Extraction with HCl and sonication addition of K4Fe(CN)g with NaOH and MeOH to oxidase thiamine to thiochrome filtration... 

Extraction is commonly carried out by hydrolysis in boiling acid such as chloridric acid or sulfuric acid. To release thiamine bonded to phosphate enzyme, hydrolysis with phosphatase, alone or together with claradiastase or takadiastase, is carried out. After the enzymatic digestion, an acid treatment is applied in order to precipitate the protein and denaturate the enzymes. Ndaw et al. [603] proved that for extraction of vitamins Bj, B, and Bg, acid hydrolysis is always superfluous if the activity of the enzymes chosen is sufficiently high. SPE or column chromatography may be used in further purification, mainly to remove excess of derivatization reagents used to convert thiamine to a highly flnorescent thiochrome derivatives. lEC may be used in purification step, as well. 

More recently [635], a unique extraction step in supplemented foods, by using hot water and a precipitation solution, following by HPLC-ELD/UV analysis has been performed for the simultaneous determination of pyridoxine, thiamine, riboflavin, niacin, pantothenic acid, folic acid, cyanoco-balamin, and ascorbic acid. The mobile phase consisting of phosphate buffer and methanol has been modified in order to perform ion-liquid chromatography by adding l-octanesulfonic acid sodium salt. Furthermore, triethylamine has been also added to improve peak symmetry. 

Reddy et al. (1983) concluded that NO inactivation of iron-sulfur proteins was the probable mechanism of botulinal inhibition in nitrite-tteated foods. In support of this conclusion, Carpenter et al. (1987) observed decreased activity of clostridial pyruvate-ferredoxin oxidoteductase and lower cytochrome c reducing ability by ferredoxin in extracts of cells treated with nitrite. NO tteatment also inhibits yeast pyruvate decarboxylase (a non-iron-sulfur protein) and py-ruvate-ferredoxin oxidoteductase from C. perfringens (McMindes and Siedler, 1988). They suggested that thiamine-dependent decarboxylation of pyruvate may be an additional site for antimicrobial effects of NO. 

The isolation of 5-formyl-2-methylpyrimidine (62) and 5-amino-2-methylpyrimidine (63) from thiamin requiring mutants of Neurospora has led to a suggestion that these may be late intermediates in the biosynthesis of (45), a contention supported in part by the demonstration of conversion of the amino compound (63) to the hydroxymethylpyrimidine (45) by yeast cell-free extracts. 

Total thiamine Milk Enzymatic hydrolysis of protein with trypsin and thiamine phosphates to thiamine with claradiastase oxidation of thiamine to thiochrome using ferricyanide (derivatization stopped with sodium sulphite) thiochrome extracted with 1-butanol Analytical Nucleosil Phenyl (150 mm, 5 fi Macherey-Nagel). Isocratic methanol + acetonitrile + isobutanol + water (80 +10+10+5 v/v/v/v). 0.7 ml/min. Fluorescence 375/430 nm (ex/em). External standardization. 76 Recoveries 95% thiamine as thiochrome from milk. 

Total thiamine Rice Extraction by refluxing Analytical Zorbax Isocratic phosphate- Online postcolumn derivati- External standardization. 77... 

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