Urine polyamines

Molins-Legua C, Campinc-Falco P, Sevillano-Cabeza A, Ped-r6n-Pons M. 1999. Urine polyamines determination using dansyl chloride derivatization in solid-phase extraction cartridges and HPLC. Analyst 124 477-482. 

Adler, H., Margoshes, M., Snyder, F. R., and Spitzer, C., Rapid chromatographic method to determine polyamines in urine and whole blood, J. Chromatogr., 143, 125, 1977. 

Aboul-Enein and Al-Duraibi (1998) employed dansyl chloride in a fluorescence assay for PUT, SP, SPD, and their acetylated derivatives by ion-pair reverse-phase chromatography. This assay could be applied to the separation of free and acetylated polyamines in biological samples. Dansyl chloride has also been used as the fluorescence reagent in the determination of polyamines in urine by Molins-Legua and colleagues (1999). Derivatization was carried out within the C18 cartridges that were used during the SPE extraction procedure. Recoveries were 80-95% for all four polyamines analyzed and the hmit of detection was 10 ng/ml. 

Ethambutol is a synthetic agent and not related to any of the other tuberculostatics. Its mechanism of action is not well understood but in actively dividing mycobacteria it appears to be an inhibitor of mycobacterial RNA synthesis. It also has effects on bacterial phosphate metabolism and on polyamine synthesis. It is an bacteriostatic agent and its main function in combination therapy is to delay the occurrence of resistance, mainly against isoniazid and rifampicin. It is well absorbed after oral administration. It is widely distributed, except to the CNS. Protein binding is about 20-30%. It is mainly excreted unchanged in the bile and urine with an elimination half-life of 3 h. Ethambutol is concentrated in erythrocytes and thus provides a depot for continuous release. 

Of the common alkyl polyamines, ethylenediamine is the most notable because of its widespread use and toxicity. Although it has a toxicity rating of only three, it can be very damaging to the eyes and is a strong skin sensitizer. The dihydrochloride and dihydroiodide salts have some uses as human and veterinary pharmaceuticals. The former is administered to acidify urine, and the latter as an iodine source. Putrescine is a notoriously odorous naturally occurring substance produced by bacteria in decaying flesh. 

Schiff bases resulting from condensation of pyridoxal with amino acids (248) and polyamines (249) occur in mammalian tissues and fluids. The Schiff base pyridoxylidenephenethylamine (132) obtained by synthesis equilibrates in aqueous solution with the carbinolamine 133 and the Schiff base 134 obtained from pyridoamine and phenylacetaldehyde (Fig. 36) (250). The material isolated from the urine of parkinsonian patients, on the basis of UV data and chromatographic comparison, is a mixture of 132-134 (251,252). 

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... 

Figure 2 demonstrates the effects of adding octane sulfonic acid to the injection solvent for a reverse-phase separation of pyridinium and deoxypyri-dinium, components of collagen, in rat urine. These polyamine containing compounds are protonated and poorly retained under the usual acidic or neutral mobile-phase conditions. The sample preparation method is simple dilution and does not afford the removal of salts from the sample. Therefore if the analytes were inadequately retained the sensitivity, as well as the method accuracy, would suffer. As the concentration of octane sulfonic acid is increased to 50 mM (Fig. 2a), both the retention time and the peak response for the analytes improve significantly. Litde improvement is obtained at higher concentrations of octane sulfonic acid and retention is not strongly dependent on the injection volume (Fig. 2b). To protect the ion source from the fouling effects of octane sulfonic acid in the injection solvent, a timed divert valve was inserted before the ion source to shunt the excess ion pair reagents to waste during the first few minutes of each injection.

Swendseid, M. E., Panagua, M., and Kopple, J. D., Polyamine concentrations in red cells and urine of patients with chronic renal failure. Life Sci. 26, 533-539 (1980). 

Thus, Amoore suggests that, if we have a specific olfactory receptor for a given odorant then that odorant might be naturally given off by the body. The sweaty odor of isovaleric acid is probably part of the foot odor and is produced by the action of skin bacteria on apocrine secretion (see below). Pyrolline, the spermous odor, has been shown to be produced by enzymatic breakdown of the polyamines in semen (40). Androst-16-en-3-one, the urinous primary odor, has axillary-like odor the related androstenol, which is found in urine, is perceived as a musky odor to some individuals (41). Both steroids are found in axillary sweat and may be formed as metabolites of apocrine secretion. Chemicals which fit the malty anosmia have not as yet been reported from human odor sources. The natural musks, such as cycloheptadecenone (civet), were first obtained from animal scent glands. 

Putrescine tetramethylenediamine, a ubiquitous Polyamine (see), formed by decarboxylation of ornithine and, in some organisms, by decarboxylation of arginine to agmatine, followed by cleavage to P. and urea. It is the precursor of spermine and spermidine in ordinary metabolism, and is essential for cell division. It accumulates during bacterial degradation of arginine. Increased protein decomposition (e. g. in cholera) leads to the appearance of P. in urine and feces. 

Rapid Chromatographic Method to Determine Polyamines in Urine and Whole Blood... 

A Gas Chromatographic Method for the Determination of Di- and Polyamines in Human Urine... 

R. W., and Waalkes, T. P. Determination of Polyamines in Urine by Gas-Liquid Chromatography Polyamines Norm. Neoplastic Growth, Proc. Symp., 1972 (Pub. 1973) 343-353 CA 79 134052h... 

Makita, M., Yamamoto, S., and Kono, M. Rapid Determination of Di- and Polyamines in Human Urine by Electron-Capture Gas Chromatography... 

Mass Fragmentographic Determination of Polyamines in the Urine of Premature Babies... 

Loading tests are not necessary for the diagnosis of amino acid transport disorders. However, pathophysiologically they may help in differentiating between different types of cystinuria by oral application of cystine and dibasic amino acids followed by analysis of plasma amino acids. The polyamines putrescine and cadaverine are produced intestinally, absorbed and excreted into the urine. Loading with dibasic amino acids will result in increased production of these polyamines [9, 10]. 

Yoshida, H. Harada, H. Nakano, Y. Nohta, H. Ishida, J. Yamaguchi, M. Liquid chromatographic determination of polyamines in human urine based on intramolecular excimer-forming fluorescence derivatization using 4-(l-pyrene)butanoyl chloride. Biomed. Chromatogr. 2004,18,687-693. 

Although the amount of DiAcSpm in urine from healthy individuals is 30 to 100 times smaller than that of major urinary polyamine components, such as Af- and A -monoacetylspennidine and monoacetylputrescine, its coefficient of variation (CV) value among 50 persons was comparable to the CV values of major urinary polyamine components. This is remarkable, considering the very low DiAcSpm content, and suggests that DiAcSpm levels in urine vary little from one individual to another. In other words, DiAcSpm is secreted in a highly controlled manner. 

We do not yet fully understand why elevated levels of urinary polyamines occur in patients with cancer, and why DiAcSpm behaves differently than other polyamine species, making it a useful tumor marker. It is unclear where DiAcSpm is produced and how it is excreted in the urine. Increased production of DiAcSpm in cancer patients may occur in two ways (1) cancer cells may generate DiAcSpm from intracellular polyamines that are produced at increased levels by actively growing cells, or (2) healthy cells or tissues may in fact be producing increased amounts of DiAcSpm using the increased levels of polyamines in the bloodstream of cancer patients as substrates. 

Monoacetylpolyamines that are reabsorbed by the renal tubular system are converted to free polyamines by acetylpolyamine oxidase (APAO) and subsequently reutilized in the body. Importantly, DiAcSpm is not reabsorbed by this route. It is likely that the total amount of DiAcSpm excreted from cells is recovered in the urine without significant loss, whereas the amount of urinary monoacetylpolyamines decreases by an unknown amount compared to that originally excreted from the cells as a result of renal reabsorption and subsequent reutilization. 

Fig. 23.1 A, A -Diacetylspermine (DiAcSpm) production and excretion in the urine. DiAcSpm levels begin to increase in cancer cells in the early stages of cancer development. In the kidney, free polyamines and monoacetylpolyamines in the primary urine are reabsorbed in the rentil proximal tubules. DiAcSpm is not reabsorbed and is eventually excreted in the urine...

Hiramatsu K, Kamei S, Sugimoto M, Kinoshita K, Iwasaki K, Kawakita M (1994) An improved method of determining free and acetylated polyamines by HPLC involving an enzyme reactor and an electrochemical detector. J Biochem (Tokyo) 115 584—589 Hiramatsu K, Sugimoto M, Kamei S, Hoshino M, Kinoshita K, Iwasaki K, Kawakita M (1995) Determination of amounts of polyamines excreted in urine demonstration of APJV -diacetylspermidine and A, Ah -diacetylspermine as components commonly occurring in normal human urine. J Biochem (Tokyo) 117 107-112 Hiramatsu K, Miura H, Kamei S, Iwasaki K, Kawakita M (1998) Development of a sensitive and accurate enzyme-Unked immunosorbent assay (ELISA) system that can replace HPLC analysis for the determination of Al, Al -diacetylspermine in human urine. J Biochem (Tokyo) 124 231-236... 

Hiramatsu K, Sakaguchi K, Fujie N, Saitoh F, Takahama E, Moriya S, Iwasaki K, Sakaguchi M, Takahashi K, Kawakita M (2013) Excretion of Al, AP -diacetylspermine in the urine of healthy individuals. Ann Clin Biochem. doi 10.1177/0004563213496978 Kabra P, Lee H, Lubich WP, Marton LJ (1986) Solid-phase extraction and determination of dan-syl derivatives of unconjugated and acetylated polyamines by reversed-phase liquid chromatography improved separation systems for polyamines in cerebrospinal fluid, urine and tissue. J Chromatogr 380 19-32... 

Kuwata G, Hiramatsu K, Samejima K, Iwasald K, Takahashi K, Koizumi K, Horiguchi S, Moriya S, Kobayashi M, Kawakita M (2013) Increase of iVAP -diacetylspennine in tissues from colorectal cancer and its liver metastasis. J Cancer Res Clin Oncol 139 925-932 Lee S, Suh J, Chung BC, Kim SO (1998) Polyamine profiles in the urine of patients with leukemia. Cancer Lett 122 1-8... 

RusseU D (1971) Increased polyamine concentrations in the urine of human cancer patients. Nat New Biol 233 144-145... 

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