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Redactorresponsabil: Prof. In order to evaluate the method of separation, purification and derivatization of testosterone in blood plasma was oxiuros causa diarrea internal standard procedure.

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Plasma testosterone varies depending on animal age. In the samples from rams aged 2 and 3 warts mouth labrador, there were no significant differences regarding warts mouth labrador concentration of testosterone, the average for rams aged 2 years was 1. Instead, in samples from rams aged 5 years have seen a sharp increase of testosterone, that reaching an averageconcentrationof9.

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Biosynthesis of this hormone occurs from cholesterol, in two ways: one is principally located in the Leydig cells of the testicles and a secondary route that takes place in adrenal. Testosterone is the hormone responsible for the appearance of primary and secondary warts mouth labrador characters and male sexual function.

They can indirectly stimulate sperm maturation. Testosterone is critical for the expression of masculine sexual behaviors in mammals.

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Minimal concentrationsoftestosteronearerequiredforexpressionofheterosexualmatingbehaviorin male sheep [Roselli et al. Testosterone secretion is correlated to external stimulants such as behavior of ewes, odor and estrous manifestation of ewes. Both sexual and pheromone production are dependent on the action of androgens [Kishk, ; Walkden Brown et al.

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A large variety of methods have been developed for determining concentrations of steroid hormones in biological warts mouth labrador. Biological tests and thinlayer chromatography were developed some time ago. More recently, in vitro tests with cell lines sensitive to hormones and immunochemistry methods were used to determine concentrations of steroid hormones. Gas chromatography and gas chromatography coupled with mass spectrometry methods were also used to detect low levels of hormones [Andrei et Lucrritiinificevol53seriaMedicinVeterinar al.

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In this study we followed, on the one hand, testing a gas chromatographic method for determining the concentration of testosterone in plasma collected from rams, and determining the relationship that exists between this concentration andageoframs. Samples of blood were collected from the jugular vein of rams, without anticoagulant, in order to separate the blood plasma. Blood plasmawasthenanalyzedaccordingtothefollowingprotocol.

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Hormonesextraction: Extraction of hormones was performed in methanol, in several stages. Thus, 10ml blood plasma was added to 10 ml methanol and internal standards.

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Obtained suspension was maintained in an ice warts mouth labrador for 10 minutes and was added 10 ml distilled water. Was homogenized again, after which the samples were passed in a hot water bath at 60C for 15 minutes. After cooling to room temperature, the mixture was centrifuge for 10 minutes at rpm. Separate liquid phase was treated, twice with hexane to remove lipids. Aqueous methanol phase was then passed into a 50 ml vial and methanol removed by evaporation 45C using liquid concentrator Eppendorf Concentrator PLUS.

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Aqueous extract thusobtainedwasfurtherseparatedandpurified. Purificationandderivatization: Column purification Varian C8SPE was conditioned previously with 2 ml methanol and 3 ml distilled water.

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Aqueous extract was passed through the column and hormones were elute with 2 ml of methanol, were collected in another tube, after which solvents warts mouth labrador removed by evaporation at 45C. The derivatization reactions was performed directly into tubes which were collected extracts purified, by adding l mixture BSTFA bistrimethylsilyl trifluoroacetamide TMCS trimethylclorosilan in relative volumes of Samples were kept for 2 hours at 60C.

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The solutions thus obtained were directly injected to gas chromatograph injectionvolumewas2 l. Prior injection, under the same conditions of analysis, of a mixture of standards has allowed us to identify the separated components by comparing the retention time.

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In Figure 1, is shown chromatogram obtained from separation of hormones in theplasmasamplecollectedfromrams. Retentiontimeoftestosteroneinplasmasampleswas For progesterone the retention time was Retention times are papillomaviruses and cancer same as those obtained when separating mixtures of standard solutions, progesterone and testosterone.

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