DE108516C - - Google Patents

Description

IMPERIAL

PATENT OFFICE.

PATENT LETTERING

CLASS 30: Health Care.

or tubercle bacilli cultures.

Patented in the German Empire on September 29, 1898.

Investigations made on the chemistry of the tubercle bacilli have shown that these bacilli have an acidic substance both in the free and in the bound state and that the salt-like compounds of this acid, far less of the acidic ones Substance itself, on the one hand extremely toxic, but on the other hand also immunizing properties get.

The acidic substance, which has the general properties of a nucleic acid, is said to be tuberculinic acid, while its salt-like compounds are said to be tuberculinic acid tuberculosamine respectively tuberculinic acid albumin.

The following is used to illustrate these substances procedure:

The bacilli are separated from the culture liquid by filtration, with sterilized liquid Water carefully, washing until all mucous or mucinous Bodies are removed. The washed bacilli are then in a desiccator over sulfuric acid sharply dried, well chopped and leached with large quantities of water. After Extraction, the bacilli are separated from the liquid by centnfugiren. From the completely clear liquid, acetic acid and mineral acids beat that as tuberculinic acid Tuberculosainin designated body, while that designated as tuberculinic acid free acid cannot be precipitated by acetic acid.

This acid is obtained from the filtrates of acetic acid precipitation with alcohol containing hydrochloric acid dejected.

Since the tuberculinic acid is bound in the substance precipitated by acetic acid Form is included, since the poisonous value of acetic acid precipitation also has the effect value by far exceeds the free acid, it could be assumed from the outset that that one arrives at high-quality preparations if one leaves the salt-like, means Acetic acid precipitated compound by removing the acid the basic pair isolated.

This result can be reached in different ways: once, by the neutral body intimately mixes with dilute mineral acids, e.g. sulfuric acid, and Shakes very thoroughly for a long time, the alkaline body being the salt of the concerned Mineral acid, ζ. B. as sulfate, goes into solution, or by the fact that one by careful Warming with dilute alkalis causes cleavage.

In the first case, the solutions turn into tuberculosamine sulphate as a completely colorless, loose substance Powder down. The best way to purify the sulfate is to add it to the appropriate one Picrat over, from which one can restore the sulphate by treatment with sulfuric acid and can remove the picric acid by shaking with ether.

Claims (3)

The free base can be set free from the sulfate by treating it with alkaline liquids, preferably with barite water. Example. 10 1 tubercle bacilli broth cultures are filtered after four to five weeks of growth, the bacilli are washed with 2 to 3 liters of sterilized water and dried with dilution in air. About 50 g of dry bacilli are obtained, which are comminuted. It must be emphasized here that bacilli dried with alcohol and defatted with ether can also be used to prepare the substances described here. After comminution, the bacilli are processed into fine emulsions with 5 liters of water in a shaker and then centrifuged for several hours. Acetic acid is added to the clear liquid poured off from the sediments until the precipitate that has formed no longer increases. The precipitate is collected on a filter and washed with water containing acetic acid. 1. Filtrate; The filtrate is under persistent Stir to double the amount of space absolute. Alcohol, of which 1 1 15 cc added strong hydrochloric acid. The precipitation is carried out on a suction filter collected, washed with alcohol and ether and dried in a desiccator (tuberculinic acid). 2. Acetic acid precipitation: The precipitate produced by acetic acid is removed from the filter taken, dissolved in as little dilute ammonia as possible, again with dilute Acetic acid precipitated, collected on a filter, with a little water and then with alcohol washed and finally dried in a desiccator. Each 10 g of the dried compound one now adds 500 cc of dilute sulfuric acid (1 per cent.) and shakes the mixture 3 to 4 hours in the shaker. It is then filtered, the shaking out with less Repeated amounts of acid once or twice, and finally the collected filtrates, possibly after previous evaporation with air dilution, with absol. Like alcohol. The precipitates are on a filter collected and dried over sulfuric acid with air dilution (tuberculosamine sulfate). If the substance obtained in this way is to be purified further, it is dissolved in warm water and if the neutral solution with sodium picrate precipitates, the well-washed one breaks down Precipitate with dilute sulfuric acid and remove the picric acid by shaking with Ether. The sulphate of tuberculosamine can pass through from the solution now obtained Alcohol can be precipitated as a completely pure phosphorus-free substance. A corresponding compound can also be obtained from the nutrient broth that has been freed from bacilli obtained by acid precipitation and shaking out the precipitate with dilute sulfuric acid will. The best acid used for this is the previously described tuberculinic acid, which are most closely related to the tuberculosamine present in the broth seems to have. Compounds of tuberculinic acid are also in the bacilli bodies after they have been leached with large quantities of liquid still present. These are connections of the relevant Acid with protein bodies. These compounds can be added to the leached residual bacilli by very dilute alkalis, e.g. B. Soda solution (0.25 proc), withdraw and from the so obtained Precipitate solutions with acetic acid or alcohol containing acetic acid (tuberculinic acid Albumin). Pa τ ε ν τ - A ν s ρ R ü c η ε: 1. Process for the preparation of a highly toxic and immunizing substance from tubercle bacilli, consisting in the fact that aqueous bacilli extracts are precipitated by acetic acid, and that of the salt of a specific one Acid, tuberculinic acid, containing precipitate by shaking with dilute Mineral acids are converted into a mineral acid salt, from which the immunizing substance passes through as a free base Treating with alkalis, preferably barythydrate, is obtained. 2. The application of the method protected by claim 1 to culture liquids. 3. In the processing of culture fluids, the replacement of acetic acid by tuberculinic acid for the purpose of isolating the immunizing substance.