Urine fractions

Amount excreted in urine Fraction excreted unchanged in urine Volume of distribution... 

Fig. 20.5 Using bioassay-guided fractionation to identify the molecular size of female sex pheromone. The dependent measure is the number of male blue crabs out of 72 tested that performed courtship stationary paddling. Stimuli were male or female urine fractionated into the indicated molecular sizes (S small <500 Da, M medium 500-1,000 Da, L large >1,000 Da, a mixture of S+M+L Mix) and positive control (unprocessed pubertal female urine = Urine) and a negative control (.SW sea water). Friedman ANOVA shows an overall difference in the responsiveness to these stimuli (P < 0.0001, n = 10). An asterisk marks stimuli that elicit significantly more males to respond compared to the sea water control (Wilcoxon post hoc tests, P < 0.05)...

The measurement of stress-related hormonal changes has been used in various ergonomic studies (see chap 16, this volume). Catecholamines (epinephrine and norepinephrine) and cortisol are the most fiequentiy used stress hormones. Because blood samples are difScult to obtain during work, analyses of urine fractions are preferred. However, these cannot indicate elTeets of short-term strain. Epinephrine indieates long-lasting mental, and sometimes emotional, strain, whereas norepinephrine acts as an indicator of physical strain. Cortisol can now be recorded, with sufficient reliability, from saliva probes. 

Appreciable quantities of alkanes are ingested by animals as constituents of plant material and also by man as artificial additives (e.g. mineral oils) of foodstuffs or as components of natural or artificially produced oils and fats. Very little is known about the details of the metabolic turnover of such compounds. As in micro-organisms, it is likely that alkanes are detoxified in animals and plants by a mixed-function oxidase which forms the alcohols that may be further metabolized or excreted in the urine as conjugates, such as glucoronides or sulphates. Most of the compounds reported were characterized (GC-MS) after extraction from urine fractions that had been incubated with the appropriate enzyme to cleave the conjugate. Some biotransformations of alkanes by animal systems (mainly in vivo) are summarized in Figure 7. 

Crump, D., Swigar, A. A., West, J. R., Silverstein, R. M., Muller-Schwarze, D., and Altieri, R., 1984, Urine fractions that release Flehmen in black-tailed deer, Odocbileus hemionus columbianus, J. Chem. Ecol., 10 203. 

Fig. 2. Biological response data, mean flehmen responses/30 minutes of three bull elephants at Washington Park Zoo, Portland. Three types of whole urine (anestrous, pre-ovulatory, and estrous) are depicted and the active fractions of estrous urine fractionated by several chromatographic methods.

Crump, D., A.A. Swigar, J.R. West, R.M. Silverstein, D. Muller-Schwartze, and R. Altieri Urine Fractions that Release Flehmen in Black-Tailed Deer, Odocoi-leus hemionus columbianus. J. Chem. Ecol. 10, 203-215 (1984). 

A subsequent technique involved pyrolysis of the metabolites after gas chromatography. The C02 C02 ratio was determined by mass fragmentog-raphy. Metabolites in a 24 h urine fraction were detected from administration of doses of C-aspirin of less than 100 ng [155,156]. 

Preparation of urine samples for chemical assay was as follows. Kaolin and acetone treated powder (see below) was dissolved in 0.2M sodium phosphate buffer, pH 5.8, and subjected to successive chromatographic separations on Amberlite CG-50 and DEAE-cellulose columns employing conditions identical with those for pituitary samples. The urine fractions isolated following CG-50 and DEAE-cellulose chromatography were designated as fractions UH and UL respectively. 

Component Identification. In preparation for gas chromatography the acidic urine fraction was reacted with diazomethane produced from DIAZALD, N-methyl-N-nitroso-p-toluenesulfonamide, Aldrich Chemical Company ( ). As the diazomethane was generated by the addition of potassium hydroxide, the ethereal diazomethane solution was distilled into an ice bath-cooled reaction flask which contained the urine acids fraction. The production of diazomethane was continued until a bright yellow color persisted In the reaction flask. The reaction mixture was allowed to sit in a hood overnight during which time the ice melted allowing the reaction mixture to come to room temperature (23°C). 

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