Biogenic amines using HPLC

Used for derivatization of biogenic amines for hplc anal, with electrochem. detn. Cryst. (EtOAc/hexane). Mp 32-34°. 

The use of HPLC to analyze biogenic amines and their acid metabolites is well documented. HPLC assays for classical biogenic amines such as norepinephrine (NE), epinephrine (E), dopamine (DA), and 5-hydroxytryptamine (5-HT, serotonin) and their acid metabolites are based on several physicochemical properties that include a catechol moiety (aryl 1,2-dihydroxy), basicity, easily oxidized nature, and/or native fluorescence characteristics (Anderson, 1985). Based on these characteristics, various types of detector systems can be employed to assay low concentrations of these analytes in various matrices such as plasma, urine, cerebrospinal fluid (CSE), tissue, and dialysate. 

Several analytical techniques including capillary electrophoresis, thin layer chromatography (TLC), GC, lEC, and HPLC, have been proposed for the determination of biogenic amines in various foods. Among these, RP-HPLC is considered the most suitable one. HPLC methods used for amine determination usually involve two steps amine extraction from the matrix and analytical determination. Depending on the complexity of food matrix and the selectivity of the final analytical determination, a further purification step may be necessary prior to the analytical determination. To ensure adequate sensitivity, a derivatization step is generally required before injection [282]. 

Table 4.4 HPLC elution gradient program used for analysis of biogenic amine-dabsyl derivatives. Solvent A) 40 mM sodium acetate, 10% dimethylformamide and 0.23% (v/v/v) triethylamine adjusted to pH 5.0 with acetic acid solvent B) 87.5% acetonitrile, 10% tert-butyl methyl ether and 2.5% water (v/v/v) flow rate l.OmL/min.

Table 4.5 HPLC eluent program used for determination of biogenic amine-diethyl ethoxymethylenemalonate (DEEMM) derivatives. Eluent A) 25 mM acetate buffer (pH 5.8) with 0.02% sodium azide eluent B) 80% acetonitrile and 20% methanol (v/v) flow rate 0.9mL/min.

Table 4.6 HPLC elution gradient program used for analysis of biogenic amine benzoyl chloride derivatives. Eluent A) 0.40 M sodium dodecyl sulfate aqueous solution buffered to pH 3.0 with 0.02M phosphate buffer, eluent B) acetonitrile.

Table 4.7 HPLC elution gradient program used for analysis of biogenic amines with the method of post-column OPA derivatization. Eluent A) 0.165 M sodium acetate (pH 5.25) and lOmM octane sulfonate eluent B) 0.2 M sodium acetate (pH 4.5) and lOmM octane sulfonate in acetonitrile/H20 (66 34) eluent C) 0.01 M sodium acetate (pH 5.25) and lOmM octane sulfonate flow rate 1 mL/min.

HPLC is widely used for the analysis of drugs, biogenic amines, steroids, and other low-molecular-weight substances in biological fluids. It has also been used with considerable success in the separation of peptides, both naturally occurring and synthetic, and some examples are collected in Table 3. Particular advantages of HPLC analyses are (a) ver-... 

Shimzu et al. investigated biogenic amines in the corpus cardiacum of Periplaneta americana using HPLC with a Neurochem neurochemical analyzer. Dopamine, tyramine, vanilic aid, and octopamine (OA) were detected in the corpus cardiacum. As amino acids, high levels of tyrosine (Tyr-4) and tryptophan were also detected at high levels. Octopamine levels in the corpus cardiacum were increased on injection of an acetone solution. 

Alberto, M. R., Arena, M. E., De Nadra, M. C. (2004). Differences between biogenic amine detection by HPLC methods using OPA and dansyl derivates. Methods in Molecular Biology, 268,481-487. 

Loullis, C. C., Hingtgen, J. N., Shea, P. A., and Aprison, M. H., 1980,7n vivo determination of endogenous biogenic amines in rat brain using HPLC and push-pull cannula, Pharmacol. Biochem. Behav. 12 959-963. 

Whilst the application of electrochemical detection in HPLC is becoming established for certain classes of compounds, such as biogenic amines [2] and plant phenolic materials [3], its use for compounds that are less easily oxidized has not been widely examined. 

A hybrid FIA/HPLC system incorporating monolithic column chromatography was developed for the determination of six opiate alkaloids (morphine, pseudomorphine, codeine, oripavine, ethylmorphine, and thebaine) and four biogenic amines (vanilman-delic acid, serotonin, 5-hydroxyindole-3-acetic acid, and homovanillic acid) in human urine, using tris(2,2 -bipyridyl)ruthenium(III) and acidic potassium permanganate CL detection (Adcock et al., 2007). This hybrid system approaches the automation and separation efficiency of HPLC while maintaining the positive attributes of FIA, such as manifold versatility, speed of analysis, and portability. 

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