2-4-dinitrophenylhydrazine and dansyl-hydrazine

The poly(styrene/acrolein) latexes (P(SA)1 and P(SA)2), differing in poly(acrolein) content, were synthesized by the emulsifier-less emulsion-precipitation polymerization of styrene and acrolein. The fraction of poly(acrolein) in the surface layer was 0.35 and 0.50, for the P(SA)1 and P(SA)2 latex, respectively. Latexes were labelled with 2,4-dinitrophenylhydrazine (DNPH), dansylhdrazine (DAH), and 1-aminopyrene (APY). Surface concentration of labels varied from 4.20.10(-7) mol m-2 (for APY label on P(SA)1 latex) to 1.54.10(-6) mol m-2 (for DNPH label on P(SA)2 latex) reflecting the fraction of polyacrolein in the surface layer and bulkiness of the label. The differences between adsorption and covalent immobilization of human serum albumin and gamma globulins onto the P(SA)2 latex and onto its derivatives labelled with the 2,4-dinitrophenyl (DNP), dansyl (DA), and pyrene (PY) groups were small. The observation conforms to the hypothesis that polyacrolein forms domains on the surface of the P(SA) latexes and that after labelling some aldehyde groups are still available for the covalent immobilization of proteins. Labelled and parent latexes were used in the model slide and turbidimetric aggregation tests for the goat anti-HSA. The fluorescent latexes, labelled with APY and DAH, and latexes labelled and with DNPH were found to be suitable for the model tests, similarly as the nonlabelled ones, however, some differences in the sensitivity, depending on the presence and the nature of labels, were noticed. The standard goat anti-HSA serum (Sigma) was detected at maximum dilution equal to 2000 in the slide test, and in the dilution region from 1.8.10(3) to 4.7.10(6) times in the turbidimetric test.

Topics: Acrolein; Adsorption; Affinity Labels; Agglutination; Binding Sites; Dansyl Compounds; Fluorescent Dyes; gamma-Globulins; Humans; Hydrazines; Latex; Mathematics; Microscopy, Electron, Scanning; Nephelometry and Turbidimetry; Particle Size; Phenylhydrazines; Polymers; Polystyrenes; Pyrenes; Reference Standards; Serum Albumin; Surface Properties

A method is described for the production of bile salt derivatives via the substituent hydroxyl groups on the steroid nucleus that permits derivatization of taurine and glycine conjugates. The method involves the oxidation of the hydroxyl groups at C-3 alpha or C-7 alpha on the nucleus by the action of hydroxysteroid dehydrogenases. The keto bile salts produced are isolated and reacted with 2,4-dinitrophenylhydrazine or Dns-hydrazine to yield ultraviolet absorbing or fluorescent derivatives, respectively. The bile salt hydrazones were separated using a reversed-phase C18 radial compression cartridge with a methanol-phosphate buffer gradient elution. Although the 3-keto derivatives of chenodeoxycholic and deoxycholic acids were not resolved, the 7-keto Dns-hydrazone of chenodeoxycholic acid was separated from 3-keto derivative of deoxycholic acid.

Topics: Animals; Bile Acids and Salts; Chromatography, High Pressure Liquid; Dansyl Compounds; Glycine; Humans; Hydrazines; Phenylhydrazines; Taurine