Thiochrome Reaction Research Articles

New chromogenic spray reagent for detection and identification of carbosulfan

A new chromogenic spray reagent has been used for detection and identification of carbosulfan in HPTLC. Alkaline hydrolysis of carbosulfan yields the sodium salt of 2,3-dihydro-2,2-dimethylbenzofuran-7-ol, which forms a purple complex with potassium ferricyanide in the thiochrome reaction. Other carbamate, organophosphorus, organochlorine, and pyrethroid insecticides and constituents of viscera (amino acids, proteins, peptides, etc.) do not react with this reagent. The detection limit of carbosulfan is ca 0.5 μg.

7] High-performance liquid chromatography determination of total thiamin in biological and food products

Publisher Summary This chapter reviews high-performance liquid chromatography (HPLC) methods for the determination of thiamin in food and blood. The HPLC methods used for thiamin are generally based on a reversed-phase column using either ultraviolet (UV) or fluorimetric detection. An external standard is prepared from a thiamin diphosphate working solution. Chromatographic peak areas are calculated and quantitation is carried out by plotting reported values against a standard calibration curve. In techniques involving fluorimetric detection, thiamin and its phosphate esters are quantitatively converted to thiochrome and thiochrome phosphate esters. This reaction can be applied to a HPLC system by using two different approaches. In the first approach, the appropriate reagent is added to the sample containing compounds and the thiochrome derivatives are chromatographed (precolumn derivatization). In the second approach, the reaction is performed after the chromatography. The thiochrome reaction is rapid as performed in highly alkaline solution, where thiochrome and its phosphate esters are stable. Reagents usually used for the alkaline oxidation are potassium hexacyanoferrate, mercuric chloride, and cyanogen bromide.

Transketolase from human erythrocytes. Purification and properties

AbstractHuman erythrocyte transketolase (sedoheptulose‐7‐phosphate: D‐glyceraldehyde‐3‐phosphate glycolaldehyde‐transferase) was purified 8200‐fold by adsorption onto hydroxylapatite, DEAE‐cellulose treatment, acetone fractionation, and chromatography on Sephadex G‐100. The purified transketolase could not be separated from glyceraldehyde‐3‐phosphate dehydrogenase, whereas the latter enzyme could be isolated in a pure state. Its homogeneity is suggested by sedimentation velocity, sedimentation equilibrium, and acrylamide electrophoresis. A molecular weight of 136000 was found. The physicochemical properties of glyceraldehyde‐3‐phosphate dehydrogenase and transketolase are very similar. A molecular weight of 136000 is suggested for transketolase, although gel filtration with Sephadex G‐100 gave only 104000 ± 10%. This discrepancy is a reflection of an interaction of transketolase with the gel filtration medium. The isoelectric point for transketolase as well as for glyceraldehyde‐3‐phosphate dehydrogenase, as determined by isoelectric focussing, was found to be around 8.5. The activity of the enzyme is close to the maximum for pH 7.5 to pH 8.6. Additions of thiamine pyrophosphate or other cofactors do not influence the activity. Several divalent cations were tested. Sulfate and phosphate inhibit transketolase approximately to 50% between 50 and 100 mM concentration. Thiamine was present in transketolase, as shown by a microbiological assay and by the thiochrome reaction. The activation energy for the formation of sedoheptulose‐7‐phosphate from xylulose‐5‐phosphate was estimated from rate measurements to be 11.2 kcal/mole in the temperature range from 5° to 55°.

Syntheses of O,S-dicholanoyl-thiamine derivatives.

Various O, S-disubstituted cholic acid esters of thiol type thiamine (I-V) were synthesized. These compounds were negative for thiochrome reaction but readily regenerate thiamine by treatment with alkali. Biological tests for these compounds are now being undertaken.

Physicochemical properties of thiaminic acids.

The physicochemical properties of thiaminic acid (TnA) and O-benzoylthiaminic acid (BTnA) were investigated and the following results were obtained.1. Each monohydrate crystal is relatively stable in drying in vacuo but by refluxing with ethanol, it is converted into anhydride.2. Ultraviolet absorption spectra are similar to those of thiamine disulfide and O-benzoylthiamine disulfide, respectively. The spectra vary according to pH, owing to the resonance in the pyrimidine ring. Each compound has isosbestic point at about 273mμ.3. Each compound is hardly soluble in organic solvents. Thiaminic acid is readily soluble in water, while BTnA is a little soluble at pH above the isoelectric point. However, it is hardly soluble in lower pH levels.4. Both of them are almost negative to the Dragendorff reagent and also negative for thiochrome reaction even after treatment with cysteine or thiosulfate. On paper chromatogram they can be detected by tert-butyl hypochlorite method as a violet spot. The Rf values of TnA and BTnA with n-butanol-acetic acid-water are 0.25 and 0.54, respectively. These values are a little higher than the corresponding compounds, thiamine (0.17) or O-benzoylthiamine (0.5).5. By paper electrophoresis, thiamine and O-benzoylthiamine migrate toward the cathode at pH 1.0-8.0, as a cationic form, while TnA and BTnA migrate toward the cathode at pH 1, do not move at pH 3.0-5.0, and toward the anode at pH sending its amphoteric character.6. From the pontentiometric titration curve, the apparent dissociation index was determined at 25±1°. pK1 (-SO3H) was 2.2 (TnA) and 2.3 (BTnA), and pK2 (-NH3+) was 6.2 (TnA) and 6.1 (BTnA). The isoelectric points are both at pH 4.2.7. By HCl-hydrolysis of TnA, formic acid and 2-methyl-4-amino-5-aminomethyl-pyrimidine were formed, whereas in the case of BTnA, benzoic acid was additionally formed.

ビタミンB<sub>1</sub>関連化合物の微生物学的研究 (第1報)

Paper chromatographic examination of thiamine crystal revealed the presence of three spots, (A), (B), and (C), besides that of thiamine. The absorption spectrum, diazo reaction, and thiochrome reaction of (B) were closely similar to those of thiamine and its hydrolysis with hydrochloric acid gave a compound showing activity against Lactobacillus fermenti-36. It is therefore assumed that (B) is acetylthiamine. The absorption spectrum of (C) in aqueous solution of pH 3.0 was similar to that of thiamine but that in 2% sodium hydroxide solution was similar to those of chloroethylthiamine and thiamine anhydride in the same solvent. Consequently, (C) is probably chloroethylthiamine or bromoethylthiamine.

Determination of Thiamine by Thiochrome Reaction

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTDetermination of Thiamine by Thiochrome ReactionMotonori Fujiwara and Kiyoo MatsuiCite this: Anal. Chem. 1953, 25, 5, 810–812Publication Date (Print):May 1, 1953Publication History Published online1 May 2002Published inissue 1 May 1953https://doi.org/10.1021/ac60077a040RIGHTS & PERMISSIONSArticle Views701Altmetric-Citations85LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (373 KB) Get e-Alerts Get e-Alerts

Absorptiometric Determinotion of Thiamine by Thiochrome Reaction (II) : Determination of Thiamine in Pharmaceutical Preparations

Absorptiometric Determinotion of Thiamine by Thiochrome Reaction (II) : Determination of Thiamine in Pharmaceutical Preparations

Determination of Thiamine by the Thiochrome Reaction, Applying Cyanogen Bromide and Alkali Reaction (III) : The Determination of Thiamine in Urine

Determination of Thiamine by the Thiochrome Reaction, Applying Cyanogen Bromide and Alkali Reaction (III) : The Determination of Thiamine in Urine

Determination of Thiamine by the Thiochrome Reaction, Applying Cyangen Bromide and Alkali Reaction (II)

Determination of Thiamine by the Thiochrome Reaction, Applying Cyangen Bromide and Alkali Reaction (II)

Absorptiometric Determination of Thiamine by Thiochrome Reaction (I) : Determination of Thiamine by Cyanogen Bromide and Alkali

Absorptiometric Determination of Thiamine by Thiochrome Reaction (I) : Determination of Thiamine by Cyanogen Bromide and Alkali

Determination of Thiamine by the Thiochrome Reaction Applying Cyanogen Bromide and Alkali Reaction (I)

Determination of Thiamine by the Thiochrome Reaction Applying Cyanogen Bromide and Alkali Reaction (I)

Vitamin B<sub>1</sub>及び諸関係化合物の研究 (第45報)

Application of acylation agent, such as benzoyl chloride, acetic anhydride, and p-nitrobenzoyl chloride, to the thiol-type vitamin B1 respectively yielded crystals of m. p. 173-174° (decomp.) (VII), m. p. 122-123° (decomp.) (VIII), and m. p. 135° (decomp.) (IX). These compounds gave negative thiochrome reaction but the reaction became positive after treatment with acid or alkali. From such facts and from their elemental analytical values, they were respectively assumed as 2-methyl-4-amino-5-[N-(3′-benzoylmer-capto-5′- benzoy-Δ2′-pentenyl-2′)]-formaminomethylpyrimidine (VII), (O, S-dibenzoyl B1) 2-methyl-4-amino-5-[N-(3′-acetylmercapto-5′-acetoxy-Δ2′-pentenyl-2′)]-formaminomethyl-pyrimidine (VIII) (O, S-diacetyl B1), and 2-methyl-4-amino-5-[N-(3′-p-nitrobenzoylmercapto-5′-hydroxy-Δ2′-pentenyl-2′)]-formaminomethylpyrimidine (IX) (S-p-nitrobenzoyl B1). These compounds were tested for the prevention of B1-deficiency in Uroloncha domestica and were found to have almost equal efficacy as vitamin B1. It was also found that the absorption of (VII) and (VIII) through the alimentary canal was better compared to vitamin B1. Since (VIII) is easily formed from vitamin B1 and vice versa, it may be assumed that, as a part of biochemical actions of vitamin B1, the pyrophosphate of S-acetyl B1 undergoes transacetylation and acetylates coenzyme A to form acetylcoenzyme A.

Vitamin B<sub>1</sub>の作用機構の研究 (第1報)

a) Iodine oxidation of Vitamin B1 hydrochloride with application of 3mol. NaOH gave “aneurin disulfide”, which containes 2mol. of crystal water and has no certain melting point. However, treatment absolute EtOH or drying for a long period of time, gives an anhydride of decompn. 179°. Acetylation with acetic anhydride gives diacetate, decompn. 155°. An attempt to synthesize this diacetate by other method to determine its structure was unsuccessful.b) Air oxidation of Vitamin B1 with application of 3mol. NaOH gave, instead of “Aneurin disulfide”, Vitamin B1 half-chloride, m.p. 125-6°, 2-methyl-4-amino-5-aminomethyl-pyrimidine and crystals of unknown structure with m.p. 224°. The same process with the application of 2mol. NaOH on Vitamin B1 hydrochloride gave crystals of m.p. 239° instead of “Aneurin disulfide”. The former crystals give positive thiochrome reaction when cysteine is applied.

Catatorulin Effect of Aneurin Disulphide

ANEURIN disulphide1, formed by opening of the thiazole ring and oxidation to the—S—S—form, does not give the thiochrome reaction, unless suitably reduced by cysteine; it was reported to have some 60 per cent of the biological activity of aneurin, when given orally to animals. I have found in catatorulin tests, by methods previously described2, with the deficient pigeon brain that it is at least as active as aneurin (Table 1).

Thiamine in Beef Muscle. A Comparison of Values of the Thiochrome Reaction Applied with and without Adsorption

Thiamine in Beef Muscle. A Comparison of Values of the Thiochrome Reaction Applied with and without Adsorption

Determination of Thiamin by Thiochrome Reaction

Determination of Thiamin by Thiochrome Reaction

CLINICAL APPLICATION OF THE THIOCHROME REACTION IN THE STUDY OF THIAMIN (VITAMIN B^1) DEFICIENCY

Article1 July 1940CLINICAL APPLICATION OF THE THIOCHROME REACTION IN THE STUDY OF THIAMIN (VITAMIN B1) DEFICIENCYHARRY J. BORSOX, M.D.HARRY J. BORSOX, M.D.Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-14-1-1 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptA Chemical method for the estimation of thiamin (vitamin B1), should have the advantages, as compared to biological assay, of speed, accuracy, cheapness, and ease of performance. The great advantage of biological assay is definite specificity. However, under well controlled conditions the uncertainties of chemical assay may become negligible.Of the several methods proposed for the chemical estimation of thiamin, the thiochrome method of Jansen1has received the most attention. Recently, the method of Prebluda and McCollum2as modified by Melnick and Field3has been shown to possess a high degree of specificity and accuracy, but the technic as described...

Quantitative Measurement of Vitamin B1 by the Thiochrome Reaction

IT has been known to us for more than a year that there is a large discrepancy between the results of biological vitamin B1 assay and of determination of aneurin by the original method of Jansen1, when both methods are applied to animal tissues. We did not like to report on these observations before having succeeded in determining vitamin B1 quantitatively by the thiochrome method. Immediately after the publication of a paper by M. A. Pyke2, however, it was clear to us that he had estimated only a very small part of the total amount of vitamin B1 actually present in the animal tissues investigated. Dr. Pyke's letter in NATURE of June 25, p. 1141, prompts us to make the following remarks.

The chemical measurement of vitamin B1 in foodstuffs and biological material by means of the thiochrome reaction

The chemical measurement of vitamin B1 in foodstuffs and biological material by means of the thiochrome reaction