Theoretical Aspects of Thin-Layer Chromatography

Brenner, M.; Niederwieser, A.; Pataki, György; Weber, Rudolf

doi:10.1007/978-3-662-01031-0_8

Cited by 10 publications

(5 citation statements)

References 51 publications

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“…(2) Determination of arginine and histidine. A volume of the hydrolysate solution corresponding to 20-40 /*g of the dry hydrolysate was spotted on to a 20 x 20 cm thin-layer chromatographic plate coated with silica gel G. The plate was developed in two dimensions using phenol:water (75:25, w/w) and w-butanol:glacial acetic acid-.water (60:20:20, v/v) (Brenner et al, 1965). After formation of the ninhydrin complexes, the spots produced by arginine and histidine were removed from the plate, eluted with 60% aqueous ethanol and diluted to 2 ml.…”

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confidence: 99%

The Component Combined Amino Acids of Some Marine Phytoplankton Species

1967

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Text-figs. 1 and 2)Gas-liquid and thin-layer chromatography have been used for the determination of 25 amino acids in the hydrolysates of 25 species of marine phytoplankton which had been grown in Erd-Schreiber medium. The general pattern of their distribution agrees with that found by earlier workers; the principal amino acids being glutamic acid, alanine, leucine and aspartic acids. Small amounts of 2-amino-M-butyric and 2-amino-wo-butyric acid together with compounds which have been tentatively identified as 4-amino-«-butyric acid and the 2-amino-and 2,5-diamino derivations of adipic acid were detected in many of the organisms. No differences were found which could be related to the taxonomy of the organisms.

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confidence: 99%

The Component Combined Amino Acids of Some Marine Phytoplankton Species

1967

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“…Protein hydrolysate, 5 µl, was loaded onto a plastic-backed silica TLC plate, which was developed with chloroform:methanol:17% ammonium hydroxide (2:2:1, v/v/v) in the first direction and with phenol:water (3:1, w/w) in the second direction (Brenner et al 1969). Amino acids were detected by ninhydrin reaction followed by fluorography (Bonner and Stedman 1978).…”

Section: Bacteria and Growth Conditionsmentioning

confidence: 99%

Biochemistry and genetics of autotrophy in Methanococcus

Whitman¹

2009

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An acetate-requiring mutant of Methanococcus maripaludis allowed efficient labeling of riboses following growth in minimal medium supplemented with [2-13 C]acetate. Nuclear magnetic resonance and mass spectroscopic analysis of purified cytidine and uridine demonstrated that the C-1 of the ribose was about 67% enriched for 13 C. This value was inconsistent with the formation of erythrose 4-phosphate (E4P) exclusively by the carboxylation of a triose. Instead, these results suggest that either (i) E4P is formed by both the nonoxidative pentose phosphate and triose carboxylation pathways or (ii) E4P is formed exclusively by the nonoxidative pentose phosphate pathway and is not a precursor of aromatic amino acids.The pentose phosphate pathway provides pentoses for nucleosides and erythrose 4-phosphate (E4P) for aromatic amino acids (AroAAs) (8, 21). In most methanogens, pentoses are formed by the oxidative pentose phosphate pathway by oxidative decarboxylation of hexoses (2,(4)(5)(6). Because extracts of Methanococcus maripaludis contained high activities for transketolase and transaldolase and lacked detectable glucose-6-phosphate and 6-phosphogluconate dehydrogenase activities, Yu et al. proposed that this organism formed pentoses by a nonoxidative pentose phosphate (NOPP) pathway (Fig. 1a) (22). Subsequent labeling studies with Methanococcus voltae and Methanococcus jannaschii confirmed some aspects of this proposed pathway (2). However, in the methanococci as well as in Methanobacterium thermoautotrophicum, Methanosphaera stadtmaniae, Methanobrevibacter smithii, and Methanospirillum hungatei, phenylalanine and tyrosine were preferentially enriched at the C-7 atom (and not at both the C-7 and C-8 atoms) following labeling with [1-13 C]pyruvate (2, 5). This pattern was not consistent with the formation of these amino acids by E4P formed by the NOPP. Thus, it was proposed that E4P was formed by carboxylation of a 3-carbon sugar and not by the NOPP pathway (Fig. 1b) (2, 5). However, there is an alternative explanation for these results. E4P may not be a precursor of AroAA biosynthesis in methanogens. This alternative can be examined in methanococci, where the enrichment of ribose is expected to be affected by the removal of E4P for AroAA biosynthesis. Thus, the labeling patterns of ribose were determined in cells of M. maripaludis grown on [2-13 C]acetate. MATERIALS AND METHODSStrain and growth conditions. For the labeling experiment, all glassware was washed overnight in 10% H 2 SO 4 and rinsed in deionized and distilled water. M. maripaludis JJ12, an acetate-requiring mutant of strain JJ1 (12), was grown in mineral medium supplemented with 1.36 g of CH 3 COONa ⅐ 3H 2 O per liter. Cultures (5 ml) were inoculated with fewer than 2.5 ϫ 10 7 cells to avoid selection for revertants and were grown under 276-kPa H 2 :CO 2 (80:20, vol/vol) at 37°C as described previously (19), except that the vitamin solution was omitted. Following overnight growth, these cultures were inoculated into 100 ml of medium containing 0.73 g o...

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“…Abgesehen von den Schwierigkeiten, welche durch reelle und scheinbare Abweichungen von der Theorie [5] z, verursacht werden (vgl. [3] [6]), bereitet die Reproduzierbarkeit der Rf-Werte gewisse Probleme.…”

Section: Neuere Untersuchungen Zurunclassified

Neuere Untersuchungen Zur Reproduzierbarkeit der Rf‐Werte in der Dünnschichtehromatographie

Pataki¹

1964

Helvetica Chimica Acta

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Dem Zusammenhang zwischen Struktur und chromatographischem Verhalten in der Papier-und Dunnschicht-Chromatographie wird in den letzten Jahren besondere Aufmerksamkeit gewidmet l). Abgesehen von den Schwierigkeiten, welche durch reelle und scheinbare Abweichungen von der Theorie [5] z, verursacht werden (vgl. [3] [6]), bereitet die Reproduzierbarkeit der Rf-Werte gewisse Probleme. In dcr Diinnschichtchromatographie wurdcn bisher folgeude Einflussfaktoren auf die Rf-Werte festgestellt : Qualitat dcs Fliessmittels und des Sorptionsmittels [7] [B] [8a], Aktivierung der Schicht [9] [lo] 11. Luftfeuchtigkeit [12]. Schichtdicke [ll] [13], Plattenformat [14], Sattigungszustand [15] [16], Laufstreckc und Entfernung des Startpunktes vom Eintauchspiegel 171 [15], Auftragmengc und Losungsmittel [lo] [14] [17], Temperatur [7] [ll] [lZ] und Technik [7]. Unter Standardbedingungen [7] [lo] [18] 3, ist die Reproduzierbarkeit der Rf-Werte in der Diinnschichtchromatographie durchaus mit jener in der Papierchromatographie vergleichbar; die durch Qualitatsschwankungen des Sorptionsmittels verursachte Storung durfte sich rechnerisch beseitigen lassen [S] 4). Zur Bereitung der Schicht wird das Sorptionsmittel in der Regel mit Wasser aufgeschlammt und mit Hilfe einer geeigneten Einrichtung, z. B. des STAHL'schen Streichgerates [lS] [21], auf Glasplatten gebracht. HORHAMMER et a,?. [22] und spater BHANDARI et al. [23] berichteten uber eine neue Methode zur manuellen Bereitung der Chromatographieschichten, bei der eine Suspension des Sorptionsmittels in Essigester [22] oder einem Gemische von Alkohol/Wasser [23] auf die Platte gegossen wird. Gegenuber den mit einem Streichgerat hergestellten Schichten fanden sie keine Rf-Unterschiede. Papierchromatographie: vgl. [l] [Z] 2, ; Diinnschichtchromatographie: vgl. [3] [4]. 2, Die Ziffern in eckigen Klammern verweisen auf das Literaturverzeichnis, S. 786. 3, GEISS et al. [12] haben kiirzlich gezeigt, dass der Einfluss der Luftfeuchtigkeit, der ubrigens auch in der Papierchromatographie beobachtet wurde [19]. ebenfalls eliminiert werden muss. 4) Aus den mathematischen Betrachtungen von RACHINSKII [ZO] geht klar hervor, dass nicht nur der Rf-Wert, sondern auch der Rb-Wert (Rf bezogen auf cine Standardsubstanz) von der Qualitat des Tragers abhangig sein kann.

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Theoretical Aspects of Thin-Layer Chromatography

Cited by 10 publications

References 51 publications

The Component Combined Amino Acids of Some Marine Phytoplankton Species

The Component Combined Amino Acids of Some Marine Phytoplankton Species

Biochemistry and genetics of autotrophy in Methanococcus

Neuere Untersuchungen Zur Reproduzierbarkeit der Rf‐Werte in der Dünnschichtehromatographie

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