BIOCHEMISTRY - Main Regulators and Biological Fluids of the Human Body - 2016

2. HORMONES

2.12. Practical Section. Study of Qualitative Reactions for Hormones

2.12.4. Qualitative Reactions for Steroid Hormones

Sex Hormones play a vital role in METABOLISM and contribute to The Development of secondary sexual characteristics. They are produced by the Gonads — the Testes and Ovaries.

The most important Female Sex Hormones include estradiol, estrone (folliculin), and estriol.

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Male Sex Hormones include androsterone and testosterone.

Experiment 7. Reaction for the phenolic group of estrone

The interaction of estrone with sulfuric acid yields a straw-yellow compound that turns orange upon heating.

To prepare an alcoholic solution of folliculin (estrone), the contents of 5 ampoules of a 0.1% oily solution of folliculin for injection are poured into a separatory funnel containing 50 ml of ethanol. The funnel is shaken, and after phase Separation, the lower (oily) layer is discarded, while the upper layer (alcoholic solution) is used for the assay.

Place 5 drops of the alcoholic solution of folliculin into a dry test tube and place it in a boiling Water bath for 5 minutes (to evaporate the alcohol). Add 1 ml of concentrated sulfuric acid (Н2SO₄) to the test tube and return it to the boiling water bath for 3 - 5 minutes. Observe the color Changes in the test tube contents.

Record your observations in the report. Draw a Conclusion regarding the ability of folliculin to react with sulfuric acid.

Experiment 8. Reaction for the 17-keto group of estrone and 17-ketosteroids in urine

17-Ketosteroids are the End products of the metabolism of Adrenocortical Hormones: 17-α-hydroxy-11-deoxycorticosterone, cortisone, hydrocortisone, and partially sex hormones. In healthy women, urinary excretion of 17-ketosteroids ranges from 6 to 15 mg per 24 hours, whereas in men it is 15 - 25 mg. The excretion of 17-ketosteroids is significantly reduced in certain pituitary disorders, Addison's disease, and testicular hypofunction in men, and increases in testicular tumors (up to 1500 mg/day), adrenal tumors (up to 2000 mg/day), and acromegaly. Under stress, the Blood concentration of corticosteroids rises, and their urinary excretion increases.

The excretion of hormones and their metabolites generally increases during physical exertion. For equivalent workloads, this process is less pronounced in well-trained individuals than in untrained ones. However, near-maximal exertion results in higher hormone secretion the higher the fitness level is. A decrease in hormone excretion is an unfavorable sign indicating overexertion and the impending depletion of the body's functional reserves.

The METHOD FOR DETERMINING 17-ketosteroids is based on their ability to form cherry-red Condensation products with m-dinitrobenzene in an alkaline medium. The color intensity is proportional to The amount of 17-ketosteroids present in the urine.

Add 5 drops of the alcoholic folliculin solution to one test tube and 5 drops of urine to another. To each, add 5 drops of a 30% sodium hydroxide solution (NаОН) and 5 drops of a 2% solution of m-dinitrobenzene - 1.3(NO₃)₂C₆Н₃ in ethanol. Mix the contents by shaking and observe the Development of the characteristic coloration in each test tube.

Note the color change of the solutions and its intensity in the report. Conclude about the interaction of the 17-Ketosteroid folliculin with m-dinitrobenzene in the presence of sodium hydroxide. Were 17-ketosteroids detected in the urine, and what does this indicate?