Manfred Wiechmann

Quantitative Bestimmung von Aminen und von Aminosuren als 1-Dimethylamino-naphthalin-5-sulfonsureamide auf Dnnschichtchromatogrammen

SummaryThe preparation of several 1-dimethyl-amino-naphthalene-5-sulphonylamides and -phenolic esters is described. The absorption spectra and the fluorescence of these amides and phenolic esters in solvents of different acidity and polarity is discussed. These DANS derivates are used for the quantitative estimation off amines and amino-acids in the range of 10−8–10−12 M. After separation on thin-layer chromatograms their determination can be carried out either by scraping of the spots followed by extracting the silica gel and measuring the intensity of fluorescence in solution or by spraying the chromatograms with triethanolamine + isopropanol (20 + 80) and direct estimation according to a method described earlier. The minimum amount of DANS derivates which can be still measured with the extraction method was found to be 1 · 10−10 mole/spot, using 5 ml cuvettes, and 5 · 10−12 mole/spot using the direct estimation method. At concentrations of about 1 · 10−9 mole/spot and 1 · 10−11 mole/spot respectively, the reproducibility for both methods is about ±3.5%.ZusammenfassungWir stellten eine Reihe von Amiden und Phenolester der 1-Dimethyl-amino-naphthalin-5-sulfonsÄure (DANS-OH) prÄparativ dar. Die Absorptionsspektren und das Fluorescenzverhalten dieser DANS-Derivate in Lösungsmitteln verschiedener AciditÄt und PolaritÄt wird diskutiert. Diese Derivate dienen zur quantitativen Bestimmung von Aminen und von AminosÄuren im 10−8 bis 10−12 Mol-Bereich. Nach ihrer Trennung auf Dünnschichtchromatogrammen können entweder die fluorescierenden Flecke ausgekratzt, das Kieselgel extrahiert und die FluorescenzintensitÄt der DANS-Derivate in Lösung bestimmt werden, oder man wertet die Chromatogramme nach dem Besprühen mit TriÄthanolamin-Isopropanol (20∶80) nach einem früher mitgeteilten Verfahren direkt aus. Die Erfassungsgrenze für die DANS-Derivate durch die Extraktionsmethode betrÄgt im Falle der Verwendung von 5 ml-Küvetten etwa 1 · 10−10 Mol/Fleck, im Falle der direkten Auswertung der Dünnschichtchromatogramme 5 · 10−12 Mol/Fleck. Die Reproduzierbarkeit ist für beide Verfahren im Konzentrationsbereich um 1 · 10−9 Mol/Fleck bzw. um 1 · 10−11 Mol/Fleck mit ±3,0 bis 3,5% gleich.

The incorporation of putrescine carbon into γ-aminobutyric acid in rat liver and brain in vivo

Summary 14C from intraperitoneally injected [1,4-14C]putrescine was incorporated into γ-aminobutyric acid of liver and brain, along with its incorporation into the polyamines spermidine and spermine and into glutamic acid. The results obtained from liver can be interpreted as follows. Formation of peripheral γ-aminobutyric acid occurs to a considerable extent on a pathway not including glutamic acid decarboxylation. γ-Aminobutyric acid supposedly plays a role as intermediary substance in the oxidation of putrescine to CO2 and of substances which are transformed into putrescine; especially in organs in which this amino acid is not connected with the functionality of inhibitory neurones. Brain γ-aminobutyric acid exhibited a lower specific activity than glutamic acid. Therefore, the present data do not prove the existence of a pathway of γ-aminobutyric acid formation parallel with the glutamic acid decarboxylation in brain.

Bestimmung von ?1 und ?1(6)-tetrahydrocannabinol in Blut, Urin und Speichel von Haschisch-Rauchern

Das aus d e m K u r v e n v e r l a u f n a c h [5] zu e rmi t t e lnde t s betr~gt e twa 7,5 h, tG, e twa 3 h, t M e twa 2 h. N a c h d e m ersten M a x i m u m verl/~uft die K u r v e Iiir ein l~ngeres In te rva l l nahe der Abszisse, bevor sie erl leut ans te ig t . Aus der zei t l ichen Differenz der be iden M a x i m a ergibt sich ein I c yon e twa 33 h ulld d a m i t ein tG~ rol l e twa 20 h. Dieser Ver lauf der t s be legt Mndeut ig die F~higke i t der H e p a t o c y t e n , nach Teilh e p a t e k t o m i e n i ch t n u r e inmalig, sonde rn wiederholt in den Zellzyklus einzutreten, wobei tG~ des zwei ten Zyklus gegeni iber IG: des e rs ten Zellzyklus, d e m In te rva l I zwischell Te i lhepa t ek tomie u n d Beg inn der D N S S y n t h e s e , n i ch t v e r k f r z t ist. Obwohl die Hepa tocy te l l berei ts einell Zel lzyklus nach der Leber resek t ion du rch l au fen haben , s ind fiir den Beg inn der zweitell S -Phase of fenbar eine e rneu te I n d u k t i o n a n d ~hlllich kompl iz ier te p r epa ra t i ve S y n t h e s e n erforderl ich wie in G 1 des ersten Zyklus .

Metabolism and autoradiographic distribution of Δ8- and Δ9 tetrahydrocannabinol in some organs of the monkey callithrix jacchus

SummaryMetabolism and autoradiographic distribution of the two isomeric tetrahydrocannabinols, (2,4-14C)-Δ8-THC and (2,4-14C)-Δ9-THC), were studied in the marmoset Callithrix jacchus. Of the two cannabinoids, Δ8-THC had a slower initial rate of biotransformation to the psychopharmacologically more potent 11-hydroxylated metabolite. This may explain the minor psychopharmacological activity of the Δ8-isomer. In glandular tissues an accumulation of unchanged Δ9-THC was observed.Autoradiography revealed characteristic label distributions in some organs 30 min after the administration of the drugs. This labelling pattern was found to be changed after a 6-hr incorporation period. The autoradiographic distribution of Δ8- and Δ9-THC appeared to be identical.

Comparative autoradiographic and metabolic study of delta8- and delta9-tetrahydrocannabinol in the brain of the marmoset Callithrix jacchus.

The label distribution in the brain of the marmoset Callithrix jacchus following intravenous application of radioactively labeled Δ8-tetrahydrocannabinol (Δ8-THC) and Δ9-THC is investigated by autoradiographic technique. Accumulations of label are observed in nuclei concerned with motor function, in the optic and acoustic pathways, and a few other structures. Of the two hydroxylated isomeres which were shown to be equally psychoactive the brain concentration of 11-OH-Δ9-THC was found to be about 3 times higher as compared with 11-OH-Δ8-THC, which may explain whyΔ9-THC is more potent than Δ8-THC. More than 90% of the radioactivity found in the brain can be attributed to the THCs and their 11-hydroxylated isomeres. Polar metabolites are almost completely absent from the brain.

[Scope and limitations of the analytical use of dansyl chloride, I: The reaction of aromatic sulfonyl chlorides with aliphatic tertiary amines: the microanalytical aspects of the Hinsberg test (author's transl)].

Contrary to the present understanding of the Hinsberg test, aliphatic tertiary amines (N,N-dimethylethylamine, bufotenin, hordenine, tropan alkaloids) are shown generally to react with aromatic sulfonyl chlorides yielding the N-substituted sulfonamide derivatives of the monodealkylated amines as the result of oxidative dealkylation of the aliphatic tertiary amines. Possible errors resulting from the indiscriminate use of either dansyl chloride, bansyl chloride or other aromatic sulfonyl chlorides for the direct analytical determination of secondary amines, secondary amino acids, noralkaloids etc., are pointed out.