Cadaverine, determination

The Derivative, 5-(biotinamido)pentylamine, contains a 5-carbon cadaverine spacer group attached to the valeric acid side chain of biotin (Thermo Fisher). The compound can be used in a carbodi-imide reaction process to label carboxylate groups in proteins and other molecules, forming amide bond linkages (Chapter 3, Section 1). However, the main use of this biotinylation reagent is in the determination of factor XHIa or transglutaminase enzymes in plasma, cell, or tissue extracts. 

Interlaboratory studies were carried out on the precision characteristics of the analytical methods used for determination of certain biogenic amines in fish and fish products, as required by German law. These included putrescine (4a), cadaverine (4b), tyramine (5) and histamine (6)78. 

Diamine oxidase (DAO) activity could be determined by first reacting cadaverine with an enzyme to produce A1-piperidine, which then was reacted with 2-aminobenzaldehyde and finally chromium(VI) oxide to form 6,7,8,9-tetrahydro-ll//-pyrido[2,l-b]quinazolin-ll-one (7). The latter was determined on an HPLC apparatus (Scheme 5) (84MI4). 

The A piperideine trimer hypothesis is supported initially by the equal incorporation of lysine and cadaverine into all three alkaloid fragments13,14 but more significantly by the incorporation of three molecules of A piperideine (1) into lupanine (10) and by the manner of this incorporation label from C-6 appeared at C-2, C-15, and, by inference, C-10, whereas C-2 label appeared at C-17, C-ll, and, by inference, C-6, consistent with the hypothesis (see Scheme 3).19 Further, approximately a third of the label was located at each of the determined sites. 

The rate of deamination is determined by the chain length and with polymethylene diamines, NH2-(CH2)n-NH2, the maximum rate occurs when n = 4 (putrescine) and n = 5 (cadaverine) and decreases to zero when n = 9 or more. At this chain length, MAO becomes active and can oxidatively deaminate the diamines. It should be noted that although MAO can deaminate both substituted and primary amines, DAO can deaminate only primary amines. 

Whether or not lysine is incorporated directly (unsymmetrically) or via cadaverine (symmetrically) was determined by feeding lysine-2-i4C. 

Another labelling agent suitable for determination by HPLC of histamine, tyramine, phenylethylamine, tryptamine, cadaverine, putrescine, spermidine and spermine in wines is 4-(4-dimethyl-aminophenylazo)benzenesulfonyl (dabsyl) chloride (see Figure 4.14) (Bockhardt et al., 1996 Romero et al., 2000). 

The most commonly applied methods for the analysis of polyamines in erythrocytes make use of amino acid analyzers and HPLC techniques. A capillary gas chromatographic method with nitrogen-phosphorous detection was applied to the simultaneous determination of 1,3-diaminopropane, putrescine, cadaverine (Cad), spermidine (Sd), and spermine (Sp) in human erythrocytes. Blood samples, collected by venipuncture into EDTA containing Venoject tubes, were subjected to the removal of plasma by centrifugation and erythrocytes were washed three times with two volumes of 0.9% NaCl. The stability of polyamines in erythrocyte suspensions was also investigated. Quantification of polyamines was done by comparing the peak-area ratio of each analyte and its internal standard with that of the standard. The polyamine samples were eluted with 0.1 M hydrochloric acid solutions. The eluate was evaporated to dryness at 120°C under a stream of air and 200 each of acetonitrile and heptafluorobutyric anhydride were added. The isolation of derivatives... 

Kim and Bjeldanes (1979) determined concentrations of cadaverine, putrescine, histamine, spermidine, and spermine in canned wholesome tuna and in canned tuna that had been implicated in an outbreak of scombroid poisoning in humans (Table II). Fresh tuna recently caught contains negligible quantities of histamine, usually less than 1 ppm (Frank et al, 1981). Fernandez-Salguero and Mackie (1987b) determined the levels of higher amines in canned tuna, mackerel, and sardine. They reported very low levels of histamine (below 0.5 mg/100 g) in all the samples analyzed, putrescine cadaverine ranged between 0 and 1.5 mg/100 g and spermidine and spermine between 0 and 2.5 mg/100 g. 

Low levels of spermidine, spermine, putrescine, cadaverine, and histamine can be determined with the TLC method developed by Spinelli et al. (1974). The method uses the fluorescent derivatives of the amines formed with dansyl chloride (5-(dimethylamino)-l-naphtalene sulfony chloride)... 

Rosier and Van Peteghem (1988) described a rapid method for the extraction, derivatization, and determination by HPLC of the 5-dimethylaminonaphthalene-l-sulphonyl (dansyl chloride) derivatives of putrescine, cadaverine, histamine, spermidine, and spermine from fish. Comparison of this procedure to earlier methods reflected considerable reduction in the time needed for sample preparation (from approx 8 to 0.5 hr) and cost (use of water and methanol instead of acetonitrile). 

Yamamoto et al. (1982) developed a quantitative method for the determination of putrescine, cadaverine, spermidine, and spermine in foods. Separation of the amines from foods was achieved by eluting through a cation-exchange resin column and then converted to their (ethyloxy) carbonyl derivatives by the reaction with ethyl chloroformate in aqueous medium before application to the gas chromatograph with a flame ionization detector. They used 1,8-diaminooctane as an internal standard and performed the separation and determination of the resulting derivatives on a 1.5% SE-30/0.3% SP-1000 on Uniport HP column (0.5 m) under the temperature-programmed condition. Staruszkiewicz and Bond (1981) developed a GLC procedure for the quantitative determination of the diamines putrescine... 

Redmond, J. W., and Tseng, A. (1979). High-pressure liquid chromatographic determination of putrescine, cadaverine, spermidine and spermine. J. Chromatog. 170,479-481. 

Rosier, J., and Van Peteghem, C. (1988). A screening method for the simultaneous determination of putrescine, cadaverine, histamine, spermidine and spermine in fish by means of high pressure liquid chromatography of their 5-dimethylaminonaphthalene-l-sulphonyl derivatives. Zeitsch. Leb. Enters. Forsch. 186, 25-28. 

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