By Alan A. Boulton, Glen B. Baker
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Additional info for Amines & Their Metabolites (NEUROMETHODS) (Neuromethods 2 Series I Neurochemistry)
7 Fg/L by Thompson and Walton (1964). The authors also state that some interference by SPD could be possible m tissues or fluids in which this polyamine is present. In 1969, Anton and Sayre proposed another modification of the original assay of Shore et al. (1959) to overcome interference from SPD and histidine and decrease variability introduced by the pH sensitivity of the OPT-HA fluorophore. The method mcludes a series of solvent extraction steps prior to the OPT condensation step. Harvey (1973) has mvestigated this method of Anton and Sayre (1969) and concluded that the overall recovery as determined by use of radiolabeled HA is 25%, not 70-75% as reported by Anton and Sayre.
1959; Sourkes and Murphy, 1961; McGeer and McGeer, 1962; Chang, 1964, Laverty and Taylor, 1968; Westermk and Korf, 1977; Hamall and Sekl, 1979). , 1976) The second prmcrpal method for formatron of fluorescent derivatives of catecholammes and metabolites IS oxidation followed by condensation with ethylenediamine (Fig. 3). Following the report of Natelson et al. (1940) that A m ammonia solution will condense with ethylenedlamine, butylamine, aniline, or o-phenylenediamme to produce fluorescent derlvatrves that can be extracted mto butyl or amyl alcohol, Well-Malherbe and Bone (1952) suggested that this reaction could be used to measure A and NA in blood after adsorptron onto alumina.
ESTIMATION OF HVA. 6 mL each) of the Tris extract are used for analysis. 2 mL) is added Potassium ferricyanide m ammonium hydroxide (1 mL) is added to each of the three tubes. 2 mL) is added to the two tubes not containing cysteine. Fluorescence IS measured at X,,, 315 nm; A,,,,, 430 nm. 3. ESTIMATION 1,2-diaminoethane Two samples (1 mL each) of the extract are employed for analysis. 1 kg) IS added to one of the samples to act as an mternal standard. After heating both tubes in the dark in a water bath (60°C for 20 mm), they are cooled m ice.
Amines & Their Metabolites (NEUROMETHODS) (Neuromethods 2 Series I Neurochemistry) by Alan A. Boulton, Glen B. Baker