Chloroacetaldehyde: fluorescence

Readily prepd. from Adenosine and chloroacetaldehyde. Fluorescent biol. probe. 

The first scientific articles from the IKhPS were submitted for publication in the early 1960s, among them being Nikolay s reports on his work in the new field. His major project in nucleotide chemistry was specific chemical modifications of heterocyclic bases. Reactions of hydroxylamine with cytidine and uridine were studied in detail and a new reagent, O-methylhydroxylamine, was proposed for modification of cytidine. These investigations aimed at the development of efficient methods for sequencing and analysis of the secondary structure of polynucleotides. Later, a reaction of chloroacetaldehyde with adenosine and cytidine was discovered and used for preparation of fluorescent polynucleotide derivatives. 

Adenosine triphosphate Derivitivised with 0.3m chloroacetaldehyde atpH4-5. Reverse phase column Spectro fluorescence [154]... 

Bromo- and chloroacetaldehyde react readily with adenine and cytosine, either free or as constituents of nucleosides and nucleotides (Figure 5)63. The etheno adducts so formed are highly fluorescent, a discovery that has made this reaction a very valuable research... 

The lack of suitable chromophores for UV-Vis detection can be circumvented by derivatization, and the same strategy can be nsed to obtain the required fluorescence of an analyte if it does not naturally fluoresce. Many flnorophores were investigated and nsed. Determination of leukocyte DNA 6-thioguanine nucleotide levels relied on derivatization with chloroacetaldehyde [25]. A post-column derivatization with )8-naphthoqninone-4-sulfonate allowed the detection of streptomycin and dihydrostreptomycin in foods [26], A post-colnmn derivatization with 0-phthalaldehyde was used for the detection of biogenic amines and polyamines in vegetable products... 

Reaction of chloroacetaldehyde with adenine, adenosine as well as its nucleotides (see Section 7.1.1.3.7.) results in the formation of the so-called etheno compounds eAde, eA, eAMP, etc. These fused purine derivatives exhibit intensive fluorescence ethenoadenosine, for example, shows an emission maximum at 415 nm (excitation 310 nm) in aqueous solution (pH 7) with a quantum yield of 0.56 (life time 20 nsec).All adenine derivatives have similar fluorescence properties the nucleotide analogs show considerable substrate activities with different kinases. 

Treatment of single-stranded nucleic acids with chloroacetaldehyde results in the formation of hydrated etheno derivatives of cytosine and adenine bases (112) and (113), which lose water slowly under physiological conditions to give the fluorescent bases 3,A -ethenocytosine (eC) and l,A -ethenoadenine (sA), respectively. DNA has been rendered fluorescent by this treatment, and the... 

Chloroacetaldehyde undergoes a highly specific reaction with adenine and related compounds to produce l,iV -ethenoadenine, a highly fluorescent derivative with an emission maximum at 410 nm (Leonard and Tolman, 1975). This method allows the specific detection of 1 pmol of adenine and related nucleosides and nucleotides (Preston,... 

In a second procedure, poly(ADP-ribose) was first separated from the bulk of the nucleic acids and proteins by dihydroxyboryl-Sepharose affinity chromatography 147,190). The isolated polymer was treated with snake venom phosphodiesterase and bacterial alkaline phosphatase to yield the nucleoside 2, l"-ribosyladenosine from internal residues. This product was then treated with chloroacetaldehyde to produce the fluorescent derivative, l,iSr -ethenoribosyladenosine, which was then separated from other derivatized residues by reversed-phase high performance liquid chromatography picomole amounts were quantified by fluorescence detection. This procedure facilitates the accurate determination of minute quantities of endogenous poly(ADP-ribose) (102, 190). Niedergang et al. (147) have also utilized a fluorimetric assay for determination of the enzymatic digestion products of the polymer, ADP-ribose, or iso-ADP-ribose. 

Next, total rat hver proteins were examined for the presence of linkages characteristic of ADP-ribosyl-cysteine. Trichloroacetic acid insoluble extracts of hver tissue were treated to remove non-covalently bound ADP-ribose and subsequently treated with mercuric ion and analyzed for ADP-ribose. Fig. 2 shows such an analysis along with control experiments. Panel A shows that analysis of an extract treated with mercuric ion exhibited a fluorescent peak that migrated at the expected elution position of etheno-ADP-ribose, the fluorescent derivative used for quantitative determination of ADP-ribose. Panel B shows the analysis in which mercuric ion was omitted from a parahel sample of hver extract. Panel C shows the result obtained when chloroacetaldehyde, which is required for the formation of the fluorescent derivative of ADP-ribose, was omitted. This control rules out the possibility of endogenously fluorescent compounds present in the ceh extract that were released by mercuric ion. Panel D shows that a small amoimt of authentic etheno-ADP-ribose added to extracts prepared as in... 

Pre-treatment of rats with phenobarbitone, which is known to increase the P450 content of microsomal enzymes, increased the mutagenic response to vinyl chloride monomer in vitro (Bartsch etal. 1975). In an aqueous solution at pH 7.4 and 37 °C the epoxy compound had a half-Ufe of 1.6 min, and its rate of hydrolysis followed a first order kinetic. Chloroethylene oxide, but not 2-chloroacetaldehyde, showed a strong alkylating activity as determined by its reaction with 4-(p-nitrobenzyl)pyridine (Malaveille etal. 1975). 2-Chloroacetaldehyde is a chemically reactive and toxic compound (Lawrence et al. 1972), and is covalently bound to cellular nucleophiles. It reacts at pH 3.5-4.5 and 37 °C with adenosine or cytidine to give fluorescent products, which have been characterised as 3- 3-D-ribofuranosyl-imidazo-(2,l-i)pur-ine or 5,5-dihydro-5-dihydro-5-oxo-5- 3-D-ribofu-... 

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