Spermidine 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. 

Kamei et al. [45] separated spermine, spermidine, putrescine, and cadav-erine in an ion-pair reversed-phase LC system and detected the hydrogen peroxide formed in the reaction catalyzed by the enzymes putrescine oxidase and polyamine oxidase with POCL. The same analytes were determined in a later study [46], together with the acetyl derivatives. The sensitive determination of uric acid, selectively converted to hydrogen peroxide by uricase, has been investigated by several authors [37, 47],... 

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. 

Dimethylamine (109), putrescine (111), and spermidine (110), isolated from various insects (Table VIII), were obtained as p,p -nitrophenylazobenzoyl, p-phenylazobenzenesulfonyl, and I-dimethylaminonaphthalene-5-sulfonyl (dan-syl) derivatives and picrates or were detected by high-performance liquid chromatography (HPLC) using the ion-exchange resin (106,343). V,V-Dimethyl-3-phenylethylamine (131) from spiders of the genus Sclerobunus (Table VIII) has been identified by mass spectral comparison with a synthetic sample (117). 

The key alkaloid of the dihydroperiphylline group is periphylline (138), on which intensive chemical and spectroscopic analysis was performed. Essential to the structure determination was the detection of spermidine in an alkali melt of tetrahydroperiphylline (139), prepared by catalytic hydrogenation of periphylline (138). Alkali hydrolysis of 138 yielded ( -cin-... 

The enzyme spermidine synthetase catalyzes the conversion of putrescine to spermidine. The separation of the substrate from the product was accomplished on a reversed phase column (Ci8) using a mobile phase of 60% methanol (v/v). The compounds in the eluent were detected by scintillation counting or by UV at 254 nm as shown in Figure 9.101 (labeled Blank). 

No mass peak at m/z 767, corresponding to the purely N3P3C15 (spermidine minus 2 H) C15P3N3 BINO structure, could ever be detected, whatever the sensitivity of the instrument. 

Hair may also be an important tool for the diagnosis and monitoring of various disease states. For example, the concentrations of polyamines (e.g., putrescine, spermidine, and spermine) in the hair may be helpful in diagnosing and assessing disease activity in women with cervical or ovarian cancer. Assessing the level of polyamines in the hair shaft is preferred to measuring them in plasma and urine because the polyamine levels can vary during the day in plasma and urine. Increased levels of porphyrins in hair have been detected in patients with porphyria... 

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... 

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