Cadaverine detection

The polyamines putrescine, cadaverine, spermidine, and spermine, which are seen at elevated levels in some victims of cancer, were separated on a Technicon (The Technicon Company Chauncey, NY) TSM Amino Acid Analyzer packed with an 8% divinylbenzene-co-polystyrene sulfonated resin with post-column ninhydrin detection.111 Amines such as ethanolamine, noradrenaline, hexamethylene diamine, methoxytryptamine, spermine, and spermidine were separated from amino acids on a DC-4A cation exchange resin.112 A similar approach, using a Beckman Model 121M amino acid analyzer equipped with an AA-20 column, was also successful.113 A Polyamin-pak strong cation exchange column (JASCO) was eluted with a citrate buffer for the detection of putrescene, spermine, cadaverine, and 1,5-diaminohex-ane from rat thymus.114 A post-column o-phthaldehyde detection system was used. 

IEC was applied to determine biogenic polyamines such as putrescine (4a), cadaverine (4b), tyramine (5), histamine (6), spermidine (38), agmatine (39) and tryptamine (40), contained in aqueous trichloroacetic extracts of leafy vegetables, such as cabbage and lettuce. A cation exchange column loaded with potassium ions and a special buffer were used. Spermidine (38) was the major amine detected in this group (7-15 Xg/g fresh weight)144. 

Okuzumi et al. (1990) investigated the relationship between microflora on horse mackerel (Trachurus japonicus) and the dominant spoilage bacteria. The results of their study showed that Pseudomonas I/II, Pseudomonas III/IV-NH, Vibrio, and Photobacterium were dominant when high levels of putrescine, cadaverine, and histamine were detected. 

Another type of electronic nose was developed at Lund University in Sweden specifically to detect the odors associated with spoiled fish. In this case the sensor uses two enzymes linked to a polymer to detect the presence of amines such as histamine, putrescine, and cadaverine. Such amines are produced by microbes as fish spoil. 

Fig. 3-164. Separation of various polyamines. — Separator column IonPac CS1 eluent see Table 3-25 flow rate 0.6 mL/min detection fluorescence after reaction with o-phthaldialdehyde injection volume 20 pL solute concentrations 4.4 ppm putrescine, 5.1 ppm cadaverine, 7.3 ppm spermidine, and 10.1 ppm spermine.

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... 

Abdel-Monem and Ohno, 1975). Shalaby (1995) described the use of TLC with a multiple development technique to resolve the dansyl derivatives of histamine, cadaverine, putrescine, phenylethylamine, tyramine, tryptamine, spermine, and spermidine from fish, cheese, and meat samples. The procedure allowed for the detection in 14 samples of as little as 5 or 10 ng of the dansyl derivatives of the amines within 2 hr. 

The dansyl derivatives, which have a napthtalene structure, are excellent derivatives for primary amines (Mietz and Karmas, 1978). They are easily formed and detected by a uv (HPLC) detector in amounts as little as 10 ng, therefore the method does not require high sensitivity or fluorometric detectors. Gradient elution improves the separation, allowing for a broad range of derivatives to be separated in a relatively short time (40 min). Use of reverse-phase microparticular (5-10 /u,m) columns can further improve the HPLC separation (Gouygou et al, 1992). Desiderio et al (1987) described the use of a reversed-phase HPLC method for the quantification of putrescine, cadaverine, spermidine, and spermine from brain extracts as the dansyl derivatives. 

The biochemistry and molecular biology of quinolizidine alkaloid biosynthesis have not been fully characterized. Quinolizidine alkaloids are formed from lysine via lysine decarboxylase (LDC), where cadaverine is the first detectable intermediate (Scheme 6). Biosynthesis of the quinolizidine ring is thought to arise from the cyclization of cadaverine units via an enzyme-bound intermediate 176). LDC and the quinolizidine skeleton-forming enzyme have been detected in chlorop-lasts of L. polyphyllus 177). Once the quinolizidine skeleton has been formed, it is modified by dehydrogenation, hydroxylation, or esterification to generate the diverse array of alkaloid products. 

We hypothesized that compounds that attract brown tree snakes could be those that are considered major components of the odor of carrion. These include cadaverine, di-methylamine, dimethyl disulfide, dimethyl sulfide, ethanethiol, trimethylamine, and putre-scine, (Eskin, Henderson Townsen, 1971 Stager, 1964). For testing, we chose odor concentrations that were easily detectable, but not overpowering, to the human nose. 

Figure 2.8 Indirect detection of N-labeled isotope by H and NMR spectroscopy (a) Incorporation of [1- C, l- N]putrescine into retronecine and (b) incorporation of [1- C, l- N]cadaverine into lupinine.

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