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The Determination of Sub-Microgram Quantities, of Amino Acids with H{sup 3}- and C{sup 1}4-Labelled 1-Fluoro-2.4 Dinitrobenzene; Determination d'acides amines en quantites inferieures au microgramme a l'aide de 1-fluoro-2,4-dinitrobenzen e marque au {sup 3}H et au {sup 14}C; Podschet submikrogrammnykh kolichestv aminokislot pri pomoshchi 2 : 4 dinitroftor'enzola, mechennogo H{sup 3} i C{sup 14}; Determinacion de cantidades de aminoacidos inferiores al microgramo por medio de 1-fluoro-2,4-dinitrobenceno marcado con {sup 3}H y {sup 14}C

Conference:

Abstract

A method of isotope derivation analysis is described for the estimation of amino acids at levels as low as 10{sup -5} {mu}M in protein hydrolysates. 1-fluoro-2.4-dinitrobenzene-H{sup 3} (G) was synthesized from bromobenzene-H{sup 3} (G), and 1-fluoro- 2.4-dinitrobenzene-C{sup 14} (G) was prepared from chlorobenzene-C{sup 14} (G). The method of analysis involves the treatment of the mixture of amino acids with the tritiated reagent (specific activity 12,8{mu}c/{mu}-equivalent) in alkaline solution followed by the addition of known activities (6000 dpm) of C{sup 14}-labelled 2.4-dinitto phenyl derivatives. After solvent extraction the reaction mixture is separated into its constituent dinitro phenyl derivatives by two-way paper chromatography, and the H{sup 3} and C{sup 14} contente of each spot are determined by combustion and gas counting. The H{sup 3} count gives a measure of the amount of DNP derivative of amino acid isolated from the original mixture, and the C{sup 14} count gives a measure pf losses ihcurred during the analysis. The results obtained from the analysis of ({mu}g of hydrolyzed insulin are in good agreement with values obtained by derivative analysis with H{sup 3} and C{sup 14}-labelled acetic anhydride and by ion exchange resin chromatography. [French] Les auteurs decrivent une methode d'analyse par derivation isotopique permettant  More>>
Authors:
Beale, D; Whitehead, J K [1] 
  1. Middlesex Hospital Medical School, London (United Kingdom)
Publication Date:
Jan 15, 1962
Product Type:
Conference
Resource Relation:
Conference: Symposium on Tritium in the Physical and Biological Sciences, Vienna (Austria), 3-10 May 1961; Other Information: 5 tabs., 12 refs.; Related Information: In: Tritium in the Physical and Biological Sciences. V. 1. Proceedings of a Symposium| 380 p.
Subject:
37 INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; AMINO ACIDS; ANHYDRIDES; CARBON 14; CHROMATOGRAPHY; DINITROPHENOL; REAGENTS; SOLUTIONS; SOLVENT EXTRACTION
OSTI ID:
22028517
Research Organizations:
International Atomic Energy Agency, Vienna (Austria)
Country of Origin:
IAEA
Language:
English
Other Identifying Numbers:
Other: ISSN 0074-1884; TRN: XA12R0269116783
Submitting Site:
INIS
Size:
page(s) 179-190
Announcement Date:
Jan 18, 2013

Conference:

Citation Formats

Beale, D, and Whitehead, J K. The Determination of Sub-Microgram Quantities, of Amino Acids with H{sup 3}- and C{sup 1}4-Labelled 1-Fluoro-2.4 Dinitrobenzene; Determination d'acides amines en quantites inferieures au microgramme a l'aide de 1-fluoro-2,4-dinitrobenzen e marque au {sup 3}H et au {sup 14}C; Podschet submikrogrammnykh kolichestv aminokislot pri pomoshchi 2 : 4 dinitroftor'enzola, mechennogo H{sup 3} i C{sup 14}; Determinacion de cantidades de aminoacidos inferiores al microgramo por medio de 1-fluoro-2,4-dinitrobenceno marcado con {sup 3}H y {sup 14}C. IAEA: N. p., 1962. Web.
Beale, D, & Whitehead, J K. The Determination of Sub-Microgram Quantities, of Amino Acids with H{sup 3}- and C{sup 1}4-Labelled 1-Fluoro-2.4 Dinitrobenzene; Determination d'acides amines en quantites inferieures au microgramme a l'aide de 1-fluoro-2,4-dinitrobenzen e marque au {sup 3}H et au {sup 14}C; Podschet submikrogrammnykh kolichestv aminokislot pri pomoshchi 2 : 4 dinitroftor'enzola, mechennogo H{sup 3} i C{sup 14}; Determinacion de cantidades de aminoacidos inferiores al microgramo por medio de 1-fluoro-2,4-dinitrobenceno marcado con {sup 3}H y {sup 14}C. IAEA.
Beale, D, and Whitehead, J K. 1962. "The Determination of Sub-Microgram Quantities, of Amino Acids with H{sup 3}- and C{sup 1}4-Labelled 1-Fluoro-2.4 Dinitrobenzene; Determination d'acides amines en quantites inferieures au microgramme a l'aide de 1-fluoro-2,4-dinitrobenzen e marque au {sup 3}H et au {sup 14}C; Podschet submikrogrammnykh kolichestv aminokislot pri pomoshchi 2 : 4 dinitroftor'enzola, mechennogo H{sup 3} i C{sup 14}; Determinacion de cantidades de aminoacidos inferiores al microgramo por medio de 1-fluoro-2,4-dinitrobenceno marcado con {sup 3}H y {sup 14}C." IAEA.
@misc{etde_22028517,
title = {The Determination of Sub-Microgram Quantities, of Amino Acids with H{sup 3}- and C{sup 1}4-Labelled 1-Fluoro-2.4 Dinitrobenzene; Determination d'acides amines en quantites inferieures au microgramme a l'aide de 1-fluoro-2,4-dinitrobenzen e marque au {sup 3}H et au {sup 14}C; Podschet submikrogrammnykh kolichestv aminokislot pri pomoshchi 2 : 4 dinitroftor'enzola, mechennogo H{sup 3} i C{sup 14}; Determinacion de cantidades de aminoacidos inferiores al microgramo por medio de 1-fluoro-2,4-dinitrobenceno marcado con {sup 3}H y {sup 14}C}
author = {Beale, D, and Whitehead, J K}
abstractNote = {A method of isotope derivation analysis is described for the estimation of amino acids at levels as low as 10{sup -5} {mu}M in protein hydrolysates. 1-fluoro-2.4-dinitrobenzene-H{sup 3} (G) was synthesized from bromobenzene-H{sup 3} (G), and 1-fluoro- 2.4-dinitrobenzene-C{sup 14} (G) was prepared from chlorobenzene-C{sup 14} (G). The method of analysis involves the treatment of the mixture of amino acids with the tritiated reagent (specific activity 12,8{mu}c/{mu}-equivalent) in alkaline solution followed by the addition of known activities (6000 dpm) of C{sup 14}-labelled 2.4-dinitto phenyl derivatives. After solvent extraction the reaction mixture is separated into its constituent dinitro phenyl derivatives by two-way paper chromatography, and the H{sup 3} and C{sup 14} contente of each spot are determined by combustion and gas counting. The H{sup 3} count gives a measure of the amount of DNP derivative of amino acid isolated from the original mixture, and the C{sup 14} count gives a measure pf losses ihcurred during the analysis. The results obtained from the analysis of ({mu}g of hydrolyzed insulin are in good agreement with values obtained by derivative analysis with H{sup 3} and C{sup 14}-labelled acetic anhydride and by ion exchange resin chromatography. [French] Les auteurs decrivent une methode d'analyse par derivation isotopique permettant d'evaluer des quantites d'acides amines aussi faibles que 10{sup -5} {mu}M dans des hydrolysats de proteine. Le 1-fluoro-2,4-dinitrobenzene-{sup 3}H (G) a ete prepare par synthese a partir du bromobenzene-{sup 3}H (U), et le 1-fluoro-2,4-dinitrobenzene-{sup 14}C (G) a partir du chlorobenzene-{sup 14}C (U). Cette methode d'analyse implique le traitement du melange d'acides amines par le reactif tritie (activite specifique: 12,8 p.c par microequivalent) en solution alcaline, suivi.de l'addition de derives du dinitro-2,4-phenyl e marque au {sup 14}C d'activite connue (6000 dpm). Apres extraction par solvant, les derives du dinitro-phenyle contenus dans le melange resultant de la reaction sont separes par chromatographic bidimensionnelle sur papier; la teneur en {sup 3}H et {sup 14}C de chaque tache est determinee combustion et comptage en phase gazeuse. Le comptage de {sup 3}H permet d'evaluei: la quantite de derives DNP (dinitro-phenyle) de l'acide amine qui a ete isolee du melange initial, et celui du {sup 14}C indique les pertes survenues durant l'analyse. Les resultats obtenus par l'analyse d'un microgramme d'insuline hydrolysee concordent avec les valeurs que donne l'analyse par derivation a l'aide d'anhydride acetique marque au {sup 3}H et {sup 14}C, et avec celles que fournit la chromatographic par echange d'ions. (author) [Spanish] La memoria describe un metodo de analisis por derivacion isotopica que sirve para determinar cuantitativamente aminoacidos en canti dades del orden de 10{sup -5} micromoles en hidrolizados de proteinas. Los autores sintetizaron el 1-fluoro-2,4-dinitrobenceno-{sup 3}H (G) a partir de bromobenceno-{sup 3}H (G) y el 1-fluoro-2,4-dinitrobenceno-{sup 14}C (G) a partir de clorobenceno-{sup 14}C (G). Segun este metodo de analisis, la mezcla de aminoacidos se trata con el reactivo tritiado (actividad especifica 12,8 {mu}c/microequivalente) en solucion alcalina, anadiendo seguidamente actividades conocidas (6000 dpm) de derivados 2,4-dinitrofenilicos marcados con {sup 14}C. Despues de la extraccion con disolventes, los derivados dinitrofenilicos presentes en la mezcla se separan por cromatografia bidimensional sobre papel y los contenidos en {sup 3}H y en {sup 14}C de cada mancha se determinan por com- bustion y recuento con ayuda de un contador de gas. El recuento del {sup 3}H indica la cantidad de derivado dinitrofenilico del aminoacido aislado de la mezcla inicial, y el recuento del {sup 14}C constituye una medida de las perdidas que se producen durante el analisis. Los resultados obtenidos al analizar 1 {mu}g de insulina hidrolizada concuerdan satisfactoriament con los valores que suministra el analisis por derivacion empleando anhidrido acetico marcado con {sup 3}H y con {sup 14}C, y con los que se obtienen mediante cromatografia de intercambio ionico. (author) [Russian] Opisyvaetsya metod izotopnogo derivatsionnogo analiza dlya opredeleniya aminokislot pri nizkom urovne v 10{sup -5} mikro-mol' v gidrolizatakh proteina. 2:4 djnitroftorbenzola-H{sup 3} (G) byl sintezirovan iz ' brombenzola-H{sup 3}(G) i 2 : 4 dinitroftorbenzola-C{sup 14}(G), kotoryj byl poluchen iz khlorbenzola-C{sup 14}(G). Metod analiza vklyuchaet obrabotku smesi amino-kislot tritirovannym reaktivom (udel'naya aktivnost' 12.8 mikro kyuri/mikro ehkvijalent) v shchelochnoj rastvore, k kotoromu prisoedinyaetsya primes' s izvestnoj aktivnost' (6000 raspadov v minutu) proizvodnykh 2 : 4 dinitrofenila , mechennogo S14. Posle rastvoryayushchej ehkstraktsii smeshannaya reaktsiya vydelyaet iz svoego sostava proizvodnye dinitrofinila dvumya putyami khromatografii na bumage i soderzhanie H''3 i C''1''4 kazhdoj kapli opredelyaetsya szhiganiem i gazovym podschetom. Podschet H''3 daet kolichestvo, ravnoe proizvodnoj DNK iz aminokisloty, otdelennoj ot osnovnoj smesi, a schet C''1''4 ukazyvaet na velichinu poter', poluchivshikhsya vo vremya analiza. Itogi, poluchennye v rezul'tate analiza 1 mikrogramma gidrolizovannogo insulina polnost'yu soglasuyutsya s velichinami, poluchennymi v rezul'tate derivatsionnogo analiza s ispol'zovaniem uksusnogo angidrida, mechennogo H{sup 3} i C{sup 14} i s pomoshch'yu khromotografii ionnogo obmena smol. (author)}
place = {IAEA}
year = {1962}
month = {Jan}
}