Price 5 kop.

STATE STANDARD

UNION SSR

TECHNICAL CONDITIONS

GOST 13805-76

Official edition

USSR STATE COMMITTEE ON STANDARDS

UDC 547.984.3: 006.354 Group H18

STATE STANDARD OF THE UNION OF THE SSR

PEPTONE DRY ENZYMATIVE FOR BACTERIOLOGICAL PURPOSES

Specifications

Dry fermentation pepton for bacteriological objects. technical conditions

GOST 13805-68

Decree State Committee and the standards of the Council of Ministers of the USSR dated April 29, 1976 No. 1002, the validity period is set

from 01.01-1977 to 01.01. 1982

Non-compliance with the standard is punishable by law

This standard applies to enzymatic peptone for bacteriological purposes, obtained from the rumen and lettuce of large cattle, sheep and goats, as well as from the stomachs of pigs using the mucous membrane of the stomachs and pancreas.

1. TECHNICAL REQUIREMENTS

1.1. Enzymatic peptone for bacteriological purposes must be manufactured in accordance with the requirements of this standard according to the technological rules approved in the prescribed manner.

1.2. Enzymatic peptone for bacteriological purposes in terms of physicochemical and biological indicators must comply with the requirements and standards specified in Table. one.

Table 1

Official publication Reprint prohibited

Reissue April 1980

© Standards Publishing, 1981

Continued tab. one

2. ACCEPTANCE RULES

2.1. Enzymatic peptone for bacteriological purposes is taken in series. A series should be understood as the amount of peptone made from the same type of raw material in one technological cycle, which has received its number and is issued with one quality document.

2.2. To control the quality of enzymatic peptone, a sample is taken from each batch of the drug in the amount of:

from a series to 100 packing units - 3%;

from a series of more than 100 packing units - 1%.

2.3. The sample is taken from packaging units taken from different places in the series.

2.4. If the test results are unsatisfactory for at least one indicator, repeated tests are carried out on it on a double number of samples taken from the same series of peptone.

The test results apply to the entire series.

3. TEST METHODS

3.1. Sampling method

3.1. L The contents of the selected packaging units are thoroughly mixed and a total sample is taken weighing no more than 500 g.

From the total sample of enzymatic peptone, four samples are isolated for analysis, each weighing at least 100 g. The selected samples are placed in clean, dry jars with ground stoppers. Corks are filled with paraffin or resin.

3.1.2. On jars with samples of enzymatic peptone stick labels indicating;

name of the drug;

manufacturing dates;

series numbers;

date of sampling;

designation of this standard.

Two samples are used for analysis, and two are left in the archives of the State Comptroller for 4 years.

3.2. Appearance, color and smell of peptone are determined organoleptically.

3.3. Determination of the concentration of hydrogen ions (pH)

3.3.1. Equipment and reagents

pH meter;

laboratory scales;

glass flask according to GOST 1770-74;

dilindr glass measured with a capacity of 100 ml in accordance with GOST 1770-74;

paper filters;

3.3.2. Conducting a test

A weighed portion of peptone weighing 1 g, weighed with an error of not more than 0.02 g, is placed in a flask and dissolved in 99 ml of boiled and cooled distilled water. A slight precipitate is allowed in the solution, which is filtered through a paper filter. In a transparent filtrate of a 1% peptone solution, the concentration of hydrogen ions (pH) is determined in accordance with the rules attached to the pH meter.

3.4. Determination of the content of insoluble impurities

3.4.1. Equipment, materials and reagents The following are used for testing: analytical balances; drying cabinet at 105°С;

cups for weighing (bottle bags) according to GOST 7148-70; desiccator according to GOST 6371-73;

glass measuring cylinder with a capacity of 100 ml according to GOST 1770-74;

cotton-gauze corks;

paper filters;

sodium oxide hydrate according to GOST 4328-77, 0.1 n. solution;

distilled water according to GOST 6709-72.

3.4.2. Conducting a test

A weighed portion of peptone weighing 5 g, weighed with an error of not more than 0.0002 g, is placed in a flask, 95 ml of distilled water are added and thoroughly mixed, achieving maximum dissolution of peptone. The prepared 5% peptone solution is filtered through a paper filter, previously dried in a bottle to constant weight and weighed with an error of not more than 0.0002 g. After filtering the peptone solution, the filter is washed with 50 ml of distilled water, placed in a bottle and dried to constant weight. at 105°C. The first weighing is carried out after drying the filter for 3 hours, subsequent weighings - after 30 minutes of drying. Drying is considered complete if the decrease in weight during two subsequent weighings does not exceed 0.0002 g.

The filtered solution of 5% peptone is diluted 5 times with distilled water. The resulting 1% peptone solution is adjusted to 0.1 N. sodium hydroxide solution to pH 7.7 and boil for 5 minutes. The solution should remain clear, without the formation of a precipitate.

After boiling, a 1% peptone solution is poured into test tubes of 7-8 ml, closed with cotton plugs and kept in an autoclave for 30 minutes at 0.15 MPa. The solution should remain clear, without the formation of a precipitate.

3.4.3. Results processing

X \u003d (t 2-nil) * 41010

where pg is the mass of peptone taken to prepare 100 g of a 5% peptone solution, g;

mp x - weight of the weighing bottle with filter before filtering, g; t 2 - weight of bottle and filter with sediment after drying, g.

3.5. Determination of moisture content

3.5.1. Equipment

For testing use;

analytical balances;

laboratory drying cabinet at 105°С;

cups for weighing (bottle bags) according to GOST 7148-70;

desiccator according to GOST 6371-73.

3.5.2. Conducting a test

A weighed portion of peptone weighing 1.5-2.0 g, weighed with an error of not more than 0.0002 g, is placed in a weighing bottle, previously dried to a constant weight. The weighed bottle is placed in an oven and dried to constant weight at 105°C.

The first weighing is carried out after 1 hour, the next - after 30 minutes of drying. Drying is considered complete if the decrease in weight during two subsequent weighings does not exceed 0.0002 g.

3.5.3. Results processing

y (w-L1\) * 1YuO

where m is the weight of the peptone sample before drying, g; rn is the weight of the peptone sample after drying, g.

3.6. Determination of the content of sulfonated ash

3.6. L Apparatus and reagents

To carry out the test, the following are used: analytical balances; heating element (tile, burner); muffle furnace;

sulfuric acid according to GOST 4204-77, concentrated.

3.6.2. Conducting a test

A portion of peptone weighing 1.0-1.5 g, weighed with an error of not more than 0.0002 g, is placed in a crucible, previously calcined to constant weight. 1 ml of concentrated sulfuric acid is added to the crucible with a peptone weight and the mixture is carefully heated on a stove until excess sulfuric acid is removed, after which the crucible is transferred to a muffle furnace and ashing is continued at a temperature of 500-600°C to a constant mass of ash.

3.6.3. Results processing

y_m\ */110(0

where m is the mass of peptone before calcination, g; m x - mass of ash after calcination, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

3.7. Determination of the content of true peptone

3.7L. Equipment and reagents

For testing use:

pipettes with a capacity of 1 and 5 ml;

glass funnels;

paper filters:

sodium oxide hydrate according to GOST 4328-77, 10% solution; copper sulfate according to GOST 4165-78, 2% solution; distilled water according to GOST 6709-72.

3.7.2. Preparing for the test Building a calibration curve

The concentration of true peptone in the solution is plotted along the abscissa axis, the value of the optical density (E) of this solution is plotted along the ordinate axis according to the data given in table. 2. Given in table. 2 values ​​of optical density (R) were obtained by measuring on a photoelectric colorimeter at a wavelength of 540 nm solutions of a specially prepared series of peptone containing a minimum of free protein and amino acids and a maximum of peptides.

Concentration of true peptone in solution, %

table 2

Optical density (I) of solution

oh dao

3.7.3. Conducting a test

True peptone is determined in a 0.5% solution of the tested peptone. To do this, a 5% peptone solution prepared in accordance with paragraph 3.4.2 is diluted 10 times with water (0.5 ml of a 5% peptone solution and 4.5 ml of distilled water).

In a test tube containing 5 ml of a 0.5% solution of the test peptone, add 0.5 ml of a 10% solution of sodium hydroxide and 0.5 ml of a 2% solution of copper sulfate. At the same time, a control is placed, for which 5 ml of water is poured into the test tube,

0.5 ml of 10% sodium hydroxide solution and 0.5 ml of 2% copper sulfate solution.

The mixture in test tubes is well mixed and after 2 min is filtered through paper filters. The color intensity is measured on a photoelectric colorimeter at a wavelength of 540-560 nm or on a spectrophotometer at a wavelength of 540 nm in cuvettes with a working length of 10 mm. The concentration of true peptone in the sample is determined by the calibration curve.

3.7.4. Results processing

V _ L M1KZO Az 05

0.5 mass of peptone (preparation) contained in 100 g of a 0.5% solution, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 5%.

3.8. Determination of total nitrogen content The total nitrogen content is determined by the Kjeldahl method or by the Kjeldahl method in combination with isothermal distillation in Conway dishes.

3.8.1. Determination of total nitrogen by the Kjeldahl method

3.8.1.1. Equipment and reagents

To carry out the test, the following are used: analytical balances;

Kjeldal flasks with a capacity of 50-100 ml according to GOST

apparatus for stripping ammonia; pipettes with a capacity of 1, 2, 5 ml;

burettes with a capacity of 25 or 50 ml with a division value of 0.05 ml; distilled water according to GOST 6709-72; sulfuric acid according to GOST 4204-77, concentrated and 0.1 n. solution;

sodium oxide hydrate according to GOST 4328-77, 30-33% and 0.1 n. solutions prepared with pre-boiled distilled water;

hydrogen peroxide according to GOST 10929-76; methyl red according to GOST 5853-51; methyl blue;

rectified ethyl alcohol according to GOST 5962-67;

methyl orange according to GOST 10816-64, 0.1% solution.

3.8.1.2. Conducting a test

1 ml of a 5% peptone solution is placed in a Kjeldal flask and 2 ml of concentrated sulfuric acid is added. A glass pear-shaped canister or a small funnel with a sealed tip is inserted into the neck of the flask and the sample is mineralized first on low heat, then the degree of heating is increased, preventing ejection of the boiling mixture into the narrow part of the flask.Mineralization is carried out in the presence of a hydrogen peroxide catalyst.The catalyst is added after the start of boiling, 0.5 ml every 15-20 minutes until the solution is completely discolored.

After cooling, the contents of the flask are transferred with 10-12 ml of distilled water to a distillation flask. The completeness of the transfer is checked by the methyl orange indicator. The last portion of flushing water should remain yellow.

20 ml of a 0.1 N solution of sulfuric acid and 2 drops of the Tashiro indicator are poured into the receiving flask of the K "jeldal apparatus. The Tashiro indicator is prepared by dissolving 0.4 g of methyl red and 0.2 g of methyl blue in 200 ml of 96% ethyl alcohol.

The stripping flask is connected to a condenser and a vaporizer. The contents of the stripping flask are neutralized by methyl orange indicator with 30-33% sodium hydroxide solution. The distillation is carried out until about 70 ml of solution is collected in the receiving flask.

The contents of the receiving flask are titrated with 0.1 N. sodium hydroxide solution until the color of the solution changes from purple to green. At the same time, a control experiment is put on the reagents, for which, instead of a peptone solution, 1 ml of distilled water is taken into the flask for mineralization.

3.8.1.3. Results processing

V _ (VK-ViKi) ■ 0.0014* 100 4 ~ 0.05

where V is the amount of 0.1 n. sulfuric acid solution poured into a receiving flask, ml;

K-correction factor to the titer 0.1 n. sulfuric acid;

Vi - the number of OD n. sodium hydroxide solution used for titration of the test or control sample, ml;

K\ - correction factor to the title of OD n. sodium hydroxide solution;

*0.05 is the mass of peptone contained in 1 ml of a 5% solution taken for analysis, g;

0.0014 - the amount of nitrogen equivalent to 1 ml of 0.1 n. sulfuric acid solution,

The final test result is the difference between the arithmetic mean of the results of two parallel determinations in experimental and control samples. Permissible discrepancies between the results of two parallel determinations should not exceed 0.2%.

3.8.2. Determination of total nitrogen content by the Kjeldahl method in combination with isothermal distillation in Conway dishes

3.8.2.1. Equipment and reagents

The following are used for the test: microburettes;

pipettes with a capacity of 1, 2, 5, 10 ml; Conway cups;

Kjeldahl flasks with a capacity of 50-100 ml according to GOST

volumetric flasks with a capacity of 100 ml according to GOST 1770-74;

vaseline according to GOST 5774-76;

hydrogen peroxide according to GOST 10929-76;

methyl red according to GOST 5853-51;

methylene blue;

rectified ethyl alcohol according to GOST 5962-67; acid sulfuric GOST 4204-77, concentrated and 0.025 N. solution;

sodium oxide hydrate (caustic soda) according to GOST 4328-77, 40% and 0.025 n. solutions;

distilled water according to GOST 6709-72.

3.8.2.2. Preparation for the test Preparation of the Tashiro indicator according to clause 3.8.1.2.

3.8.2.3. Conducting a test

5 ml of a 5% peptone solution are mineralized with heating in a Kjeldahl flask with 10 ml of sulfuric acid (concentrated) until a colorless solution is obtained. 0.5 ml of perhydrol is used as a catalyst. After mineralization, the sample was quantitatively transferred into a 100 ml volumetric flask and made up to the mark with distilled water. Pour exactly 2 ml of 0.025 N into the inside of the Conway dish. sulfuric acid solution. To prevent the liquid used from spreading, the cup is set obliquely. Then close the lid, smeared with petroleum jelly, leaving a small gap, through which 1 ml of the test solution after mineralization is pipetted into the outer part of the dish. The cup is moved so that there is a gap 5-6 mm wide on the opposite side, and 4 ml of a 40% alkali solution is quickly poured through it. Close the cup tightly with a lid, gently mix the contents in a circular motion and leave for 15-18 hours. Excess acid not bound to ammonia

titrated with 0.025 N. sodium hydroxide solution according to the Tashiro indicator. In parallel, a control sample is placed, for which distilled water is placed in the outer part of the Conway cup instead of the test solution.

3.8.2.4. Results processing

v 0,G0035(V-V\)K* 100 * 100* Kyu ^5--- ’

where V is the amount of 0.025 n. sodium hydroxide solution used for titration of the control sample, ml;

V x - the amount of 0.025 n. sodium hydroxide solution used for titration of the test sample, ml;

K - correction factor to the titer 0.025 n. sodium hydroxide solution;

0.00035 is the amount of nitrogen equivalent to 1 ml of 0.025 N. sodium hydroxide solution, g;

5-mass of peptone taken to obtain a 5% solution, g;

5 - the amount of the drug taken for mineralization, ml; 1 - the amount of the test solution, ml.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

3.9. Determination of nitrogen content of amino groups of amino acids and lower peptides

3.9. L Apparatus and reagents

laboratory glasses according to GOST 10394-72; pipettes with a capacity of 1, 2 and 20 ml;

burettes with a capacity of 5 and 10 ml with a division value of 0.05 ml; formalin technical according to GOST 1625-75; sodium oxide hydrate according to GOST 4328-77, 0.1 n. solution; sulfuric acid according to GOST 4204-77, 0.1 N. solution; phenolphthalein according to GOST 5850-72, 1% alcohol solution; distilled water according to GOST 6709-72.

3.9.2. Preparing for the test

Immediately before work, the formol mixture is prepared. To do this, add 1 ml of a 1% solution of phenolphthalein and 0.1 N of formalin to 50 ml of formalin. with a solution of sodium hydroxide, bring the color of the mixture to slightly pink.

3.9.3. Conducting a test

To 2 ml of a clear 5% peptone solution prepared in accordance with paragraph 3.4.2, add 18 ml of distilled water and adjust the pH of the mixture to 7.0 potentiometrically 0.1 N. a solution of sodium hydroxide or sulfuric acid. Then, 2 ml of the formol mixture is added and, with constant stirring, titrated potentiometrically with 0.1 N. sodium hydroxide solution to pH 9.2.

At the same time put the control on the reagents, for which instead of 2 ml of 5% peptone take 2 ml of distilled water.

3.9.4. Results processing

y _ (V-VQK-0WM-100

where V is the amount of 0.1 n. sodium hydroxide solution used for titration of the test sample, ml;

V\ - the amount of 0.1 n. sodium hydroxide solution used for titration of the control sample, ml;

K - correction factor to the titer of 0.1 n. sodium hydroxide solution;

0.0014 - the amount of nitrogen equivalent to 1 ml of 0.1 n. sodium hydroxide solution, g;

0.1 mass of peptone contained in 2 ml of 5% peptone, g

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

3.10. Determination of free protein content

3.10.1. Equipment and reagents

For testing use:

photoelectric colorimeter;

pipettes with a capacity of 5 and 10 ml;

glass test tubes according to GOST 10515-75;

trichloroacetic acid 20% solution.

3.10.2. Conducting a test

To 5 ml of the filtrate of a 5% peptone solution prepared in accordance with paragraph 3.4.2, add 5 ml of a 20% solution of trichloroacetic acid and mix well. After 5 minutes, the optical density of the mixture is determined on a photoelectric colorimeter at a wavelength of 630 nm (red light filter) in cuvettes with a working length of 10 mm against the same peptone solution diluted 1: 1 with distilled water.

3.10.3. Results processing

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.05%.

3.11. Determination of chloride content

3.11.1. Equipment and reagents

For testing use:

technical scales with a weighing error of not more than 0.01 g; heating element (burner, stove); pipettes with a capacity of 1, 2, 10, 20 and 25 ml; burettes with a capacity of 25 ml with a division value of 0.05 ml; laboratory glasses according to GOST 10394-72; distilled water according to GOST 6709-72; silver nitrate according to GOST 1277-75, 0.1 n. solution; ferroammonium alum according to GOST 4205-77, saturated solution;

potassium thiocyanate according to GOST 4239-77 or ammonium thiocyanate according to ST SEV 222-75, 0.1 n. solution;

nitric acid according to GOST 4461-77, concentrated.

3.11.2. Conducting a test

To 20 ml of a 1% peptone solution prepared in accordance with paragraph 3.4.2, add 30 ml of distilled water, 1 ml of concentrated nitric acid and 10 ml of 0.1 i. silver nitrate solution. The solution is heated to boiling and quickly cooled by immersing the beaker of the solution in a vessel of water at room temperature. After cooling the mixture, add 2 ml of a saturated solution of iron ammonium alum (indicator) and titrate with 0.1 N. a solution of potassium thiocyanate (or ammonium thiocyanate) until a brick-red color appears.

3.11.3. Results processing

Y (VK-ViKi) 0.003545-100 7 02

where V is the amount of 0.1 n. a solution of silver nitrate, taken into a beaker with a sample, ml;

K - correction factor to the titer of 0.1 n. silver nitrate solution;

V x - number 0.1 and. solution of potassium thiocyanate or ammonium thiocyanate used for sample titration, ml;

K\ - correction factor to the titer of 0.1 n. a solution of potassium thiocyanate or ammonium thiocyanate;

0.003545 - the amount of chlorides (chlorine ion), equivalent to 1 ml of OD n. silver nitrate solution, g;

0.2 is the mass of peptone contained in 20 ml of a 1% solution, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.1%.

3.12. Salt determination heavy metals

3.12.1. Equipment and reagents

For testing use:

technical scales with a weighing error of not more than 0.01 g; heating element (tile, burner); muffle furnace;

porcelain crucibles according to GOST 9147-73;

measuring cylinders with a capacity of 10 ml according to GOST 1770-74; sulfuric acid according to GOST 4204-77, concentrated; ammonium acetate according to GOST 3117-78, saturated solution;

distilled water according to GOST 6709-72; acetic acid according to GOST 61-75, 10% solution; sodium sulfide according to GOST 2053-77, saturated solution; sodium oxide hydrate according to GOST 4328-77, 10% solution; litmus or universal indicator.

3.12.2. Conducting a test

A weighed portion of peptone weighing 1 g, weighed with an error of not more than 0.02 g, is placed in a crucible, moistened with 2 ml of concentrated sulfuric acid and carefully ashed first on a stove or burner, and then in a muffle furnace at 500-600 ° C until a homogeneous ash is obtained light grey.

After cooling the crucible, the ash is poured into 1 ml of a saturated solution of ammonium acetate and diluted with 5 ml of distilled water. The contents of the crucible are filtered into the cylinder through a paper filter. The crucible and filter are washed with distilled water, bringing the volume of the solution in the cylinder to 10 ml.

The reaction of the filtrate in the cylinder is brought to neutral or slightly acidic by litmus by adding dropwise 10% acetic acid or 10% sodium hydroxide solution. Then add 1 ml of 10% acetic acid, 5 drops of a saturated solution of sodium sulfide and mix. After 5 minutes, the solution should not have a precipitate and a yellow color of the solution.

3.13. Determination of the presence of free indole

3.13.1. Equipment and reagents

For the test, the following are used: glass test tubes according to GOST 10515-75; pipettes with a capacity of 1, 2 and 10 ml; rubber cone plugs according to GOST 7852-76;

amyl alcohol;

potassium nitrite according to GOST 4144-79, 0.1% solution; sulfuric acid according to GOST 4202-75, concentrated.

3.13.2. Conducting a test

Add 2 ml of amyl alcohol to a test tube with 10 ml of 1% peptone solution, close the tube with a cork, shake well and add 1 ml of 0.1% potassium nitrite solution and 1 ml of concentrated sulfuric acid. The mixture is gently stirred and allowed to settle.

No pink coloration should appear in the alcohol layer. The appearance of a pink coloration of the alcohol layer indicates the presence of free indole in the peptone.

3.14. Determination of the ability to support microbial growth

3.14.1. Equipment, materials and reagents

technical scales with a weighing error of not more than 0.01 g; pH meter;

thermostat at 37°C; autoclave;

photoelectric colorimeter; test tubes according to GOST 10515-75; cotton-gauze corks; micropipettes with a capacity of 0.2 ml; pipettes with a capacity of 10 ml; burettes with a capacity of 25 or 50 ml; paper filters;

nutrient media (meat water according to GOST 20729-75); microbiological agar according to GOST 17206-71 or other microbiological agar;

sodium chloride according to GOST 4233-77.

3.14.2. Preparing for the test

Meat-peptone broth (MPB) is prepared in accordance with the requirements of GOST 20730-75 and meat-peptone agar (MPA). To prepare the MPB, 1% of the tested peptone, 0.5% sodium chloride are added to the meat water and the pH of the medium is set to 7.4.

To prepare MPA, 1% of the test peptone, 0.5% sodium chloride, 2% microbiological agar are added to meat water and the pH of the medium is set to 7.4.

MPB and MPA are filtered, poured into test tubes in 8 ml, closed with cotton gauze stoppers and parchment caps and sterilized in an autoclave for 30 minutes at 0.15 MPa.

3.14.3. Conducting a test

Test tubes with sterile BCH and oblique MPA are inoculated with daily control cultures. Inoculation of each culture is carried out with a micropipette of 0.2 ml in three test tubes with MPB and MPA.

Incubation is carried out for 24 hours at 37-38°C.

The typical growth of daily cultures is determined on the MPA visually and under a microscope. Growth should be typical for each microorganism stamp. The growth intensity is evaluated in the BCH on a photoelectrocolorimeter at a wavelength of 620-640 nm in a cuvette with a working length of 5 mm.

4. PACKAGING, PACKAGING, MARKING, TRANSPORTATION

AND STORAGE

4.1. Peptone is packaged with a net weight of 250 to 1000 g in cans in accordance with GOST 5981-71 or, by agreement with the consumer, with a net weight of 30 kg in plywood barrels in accordance with GOST 5958-79 with a polyethylene film lining in accordance with GOST 10354-73. The cans are hermetically sealed.

4.2. A label is attached to the jar indicating: the name of the enterprise * manufacturer; name of the drug;

series numbers; net weight; storage conditions; expiration date;

4.3. Banks with peptone are packed in plank non-separable boxes for canned food* in accordance with GOST 13358-72 or in corrugated cardboard boxes for canned food and food liquids in accordance with GOST 13516-72.

4.4. Each box and barrel is marked in accordance with GOST 14192-77 with additional data:

name of the manufacturer;

name of the drug;

series numbers;

manufacturing dates;

gross and net weights;

storage conditions;

expiration date;

designations of this standard.

A warning sign "Afraid of dampness" is applied to each box or barrel.

4.5. Peptone is stored in a dry room at a temperature not exceeding 30°C. The shelf life of peptone in hermetically sealed jars is 3 years, in plywood barrels - 1 year from the date of manufacture.

4.6. Peptone is transported by all modes of transport.

4.7. In case of violation of the sealing of the container, peptone is stored in a tightly closed vessel or plastic bag in a dry room at a temperature of 0 to 8 ° C for no more than 1 month.

Amendment No. 1 to GOST 13805-76 Dry enzymatic peptone for bacteriological purposes. Specifications

By the Decree of the State Committee of the USSR on Standards dated 31L2 81 Ai 5930, the introduction period was established

Put the code under the name of the standard: OKP 93 8881.

Throughout the text of the standard, replace the unit of measurement: ml per cm *.

Clauses 2.2, 2.3, 3.1.1. Replace the words: “packaging units” with “cans”.

Paragraph 2.2 shall be supplemented with the words: "from the series in barrels - each barrel".

Clause 2.3. Replace the words: “select” with “make up”.

Clause 2.4. Change the words: “from the same serine peptone” to “from the same sample”.

Paragraph 3.1.1 shall be supplemented with the words: "When sampling from barrels, peptone weighing at least 500 g is taken. Samples for analysis are isolated as described above."

Clauses 3.4.1, 3.7.1, 3.8.1.1, 3.8.2.1, 3.9.1, 3.12.1. Replace reference: GOST 4328-66 with GOST 4328-77.

Clauses 3.6.1, 3.8.1.1, 3.8.2.1, 3.9.1, 3.12.1. Replace reference: GOST 4204-66 and GOST 4204-77.

Paragraphs 3.7.1, 3.8.1.1, 3.8.2.1, 3.9.1, 3.10.3, 3.11.1, 3.13.1, 3.14.1 regarding pipettes and burettes shall be supplemented with the reference: "according to GOST 20292-74".

(Continued see page 170J

Paragraph 4.1 after the words "plywood barrels" shall be supplemented with the words: "type II"; replace the reference: GOST 5958-70 with GOST 5958-79.

Clause 4.4. Replace the words: “Each box and barrel is marked in accordance with GOST 14192-71 with additional data” for “Transport marking in accordance with GOST 14192-77 with additional data”; “name of the manufacturer” to “name of the manufacturer and its trademark”; "precautionary" to "manipulative".

Paragraph 4.6 shall be supplemented with the words: “with observance of the rules for the carriage of goods* operating on this type of transport.

Reinforcement of packages in transport packages when transporting goods by rail according to GOST 21929-76 and GOST 15901-70.

(IUS No. 3 1982)

Amendment No. 2 to GOST 13805-76 Dry enzymatic peptone for bacteriological purposes. Specifications

By the Decree of the State Committee of the USSR for Standards dated 04.11.86 No. 3382, the introduction period was established

Clause 1.2. Table 1. Column "Name of the indicator". Replace the words: “Free protein content, optical density, no more” with “Presence of free protein, optical density, no more”; "Content of salts of heavy metals" to "Presence of salts of heavy metals"; "Content" to "Mass fraction" (7 times).

Clauses 3.4, 3.4.3, 3.5, 3.5.3, 3.6, 3.6.3, 3.7, 3.7.4, 3.8, 3.8.1, 3.8.1.3, 3.8.2, 3.8.2.4, 3.9, 3.9.4, 3.11 , 3.11.3. Replace the word: “kept woman” with “mass fraction”.

Paragraph 3.7.1 shall be stated in a new wording:

(Continued see p. 290)

"3.7.1, Apparatus and reagents

For testing use:

photoelectric colorimeter or spectrophotometer;

desktop centrifuges type OPn-8-U4,2,

pipettes with a capacity of 1 and 5 cm 3 according to GOST 20292-74;

glass test tubes according to GOST 25336-82;

sodium oxide hydrate according to GOST 4328-77, 10% solution;

copper sulfate according to GOST 4165-78, 2% solution;

distilled water according to TQCT 6709-72.

Clause 3.7 3. The last paragraph should be reworded: “The mixture in test tubes is well mixed and centrifuged for 10 minutes at a rotation frequency of 5000-6000 min-1. The color intensity is measured on a photoelectric colorimeter at a wavelength of 540-560 nm or on a spectrophotometer at a wavelength of ? 540 nm in cuvettes with a working length of 10 mm. The concentration of true peptone in the sample is determined by the calibration curve,

Item EVIL. Replace the words: “Determination of free protein content” x “Determination of free protein” *

Clause 3.11 L. Delete the words: “according to ST SEV 222-75”,

(IUS No. 2 1987)

Editor V. S. Babkina Technical editor F. I. Shraibshtein Proofreader E. V. Mityai

Rented in emb. 10/14/80 Signed. in to lay down. 02/02/81 1.0 p. l. 1.12 uch, -ed. l. Tyr. 4000 Price 5 kop.

Order "Badge of Honor" Publishing House of Standards, Moscow, D-557, Novopresnensky per., 3. Vilnius Printing House of Publishing House of Standards, st. Mindaugo, 12/14. Zach. 5123

BASIC JV UNITS

DERIVATED UNITS WITH AND HAVING OWN NAMES

* These two expressions include, along with the basic units. SI, additional

unit-with haradizi

GOST 13805-76*

Group H18

STATE STANDARD OF THE UNION OF THE SSR

PEPTONE DRY ENZYMATIVE FOR BACTERIOLOGICAL PURPOSES

Specifications

Dry fermentation peptone for bacteriological purposes.
Specifications

Introduction date 1977-01-01

By the Decree of the State Committee of Standards of the Council of Ministers of the USSR dated April 29, 1976 N 1002, the introduction period was set from 01.01.77

The validity period was removed by decision of the Interstate Council for Standardization, Metrology and Certification (IUS 5-92)

INSTEAD OF GOST 13805-68

* REPUBLICATION (August 1995) with Amendments No. 1, 2, approved in December 1981, November 1986 (IUS 3-82, 2-87).


This standard applies to enzymatic peptone for bacteriological purposes, obtained from the rumen and lettuce of cattle, sheep and goats, as well as from the stomachs of pigs using the mucous membrane of the stomach and pancreas.

1. TECHNICAL REQUIREMENTS

1. TECHNICAL REQUIREMENTS

1.1. Enzymatic peptone for bacteriological purposes must be manufactured in accordance with the requirements of this standard according to the technological rules approved in the prescribed manner.

1.2. Enzymatic peptone for bacteriological purposes in terms of physicochemical and biological indicators must comply with the requirements and standards specified in Table 1.

Table 1

2. ACCEPTANCE RULES

2.1. Enzymatic peptone for bacteriological purposes is taken in series. A series should be understood as the amount of peptone made from the same type of raw material in one technological cycle, which has received its number and is issued with one quality document.

2.2. To control the quality of enzymatic peptone, a sample is taken from each batch of the drug in the amount of:

from a series to 100 cans - 3%;

from a series of more than 100 cans - 1%;

from a series in barrels - each barrel.

2.3. The sample is made up of jars taken from different places in the series.

2.4. In case of unsatisfactory test results for at least one indicator, repeated tests are carried out on it on a double number of samples taken from the same peptone sample.

The test results apply to the entire series.

2.2-2.4.

3. TEST METHODS

3.1. Sampling method

3.1.1. The contents of the selected jars are thoroughly mixed and a total sample is taken weighing no more than 500 g.

From the total sample of enzymatic peptone, four samples are isolated for analysis, each weighing at least 100 g. The selected samples are placed in clean, dry jars with ground stoppers. Corks are filled with paraffin or resin.

When sampling from barrels, peptone weighing 500 g is taken. Samples for analysis are isolated as described above.

(Changed edition, Rev. N 1).

3.1.2. Enzymatic peptone sample jars are labeled with:



name of the drug;

manufacturing dates;

series numbers;

date of sampling;



Two samples are used for analysis, and two are left in the archives of the State Comptroller for 4 years.

3.2. Appearance, color and smell of peptone are determined organoleptically.

3.3. Determination of the concentration of hydrogen ions (pH)

3.3.1. Equipment and reagents



pH meter;

laboratory scales;

glass flask according to GOST 1770-74;

GOST 1770-74;

paper filters;

distilled water according to GOST 6709-72.

3.3.2. Conducting a test

A weighed portion of peptone weighing 1 g, weighed with an error of not more than 0.02 g, is placed in a flask and dissolved in 99 cm 3 of boiled and cooled distilled water. A slight precipitate is allowed in the solution, which is filtered through a paper filter. In a transparent filtrate of a 1% peptone solution, the concentration of hydrogen ions (pH) is determined in accordance with the rules attached to the pH meter.

3.4. Determination of the mass fraction of insoluble impurities

(Changed edition, Rev. N 2).

3.4.1. Equipment, materials and reagents

For testing use:

analytical balances;

drying cabinet at 105 °C;

GOST 25336-82;

desiccator according to GOST 25336-82;

glass measuring cylinder with a capacity of 100 ml in accordance with GOST 1770-74;

cotton-gauze corks;

paper filters;

sodium oxide hydrate according to GOST 4328-77, 0.1 n. solution;

distilled water according to GOST 6709-72.

3.4.2. Conducting a test

A weighed portion of peptone weighing 5 g, weighed with an error of not more than 0.0002 g, is placed in a flask, 95 cm3 of distilled water are added and thoroughly mixed, achieving maximum dissolution of the peptone. The prepared 5% peptone solution is filtered through a paper filter, previously dried in a bottle to constant weight and weighed with an error of not more than 0.0002 g. After filtering the peptone solution, the filter is washed with 50 cm3 of distilled water, placed in a bottle and dried to constant weight. at 105 °C. The first weighing is carried out after drying the filter for 3 hours, subsequent weighings - after 30 minutes of drying. Drying is considered complete if the decrease in weight during two subsequent weighings does not exceed 0.0002 g.

The filtered solution of 5% peptone is diluted 5 times with distilled water. The resulting 1% peptone solution is adjusted to 0.1 N. sodium hydroxide solution to pH 7.7 and boil for 5 minutes. The solution should remain clear, without the formation of a precipitate.

After boiling, a 1% peptone solution is poured into test tubes of 7-8 cm, closed with cotton plugs and kept in an autoclave for 30 minutes at 0.15 MPa. The solution should remain clear, without the formation of a precipitate.

3.4.3. Results processing

The mass fraction of insoluble impurities () as a percentage is calculated by the formula

where is the mass of peptone taken to prepare 100 g of a 5% peptone solution, g;

- mass of weighing bottle with filter before filtration, g;

- weight of bottle and filter with sediment after drying, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.1%.

(Changed edition, Rev. N 2).

3.5. Determination of the mass fraction of moisture

(Changed edition, Rev. N 2).

3.5.1. Equipment

For testing use:

analytical balances;

laboratory drying cabinet at 105 °C;

cups for weighing (bottle bags) in accordance with GOST 25336-82;

desiccator according to GOST 25336-82.

3.5.2. Conducting a test

A weighed portion of peptone weighing 1.5-2.0 g, weighed with an error of not more than 0.0002 g, is placed in a weighing bottle, previously dried to a constant weight. The weighed bottle is placed in an oven and dried to constant weight at 105°C.

The first weighing is carried out after 1 hour, the next - after 30 minutes of drying. Drying is considered complete if the decrease in weight during two subsequent weighings does not exceed 0.0002 g.

3.5.3. Results processing

The mass fraction of moisture () as a percentage is calculated by the formula

where is the mass of the peptone sample before drying, g;

is the weight of the peptone sample after drying, g.



(Changed edition, Rev. N 2).

3.6. Determination of the mass fraction of sulfonated ash

(Changed edition, Rev. N 2).

3.6.1. Equipment and reagents

For testing use:

analytical balances;

heating element (tile, burner);

muffle furnace;

porcelain crucibles according to GOST 9147-80;

sulfuric acid according to GOST 4204-77, concentrated.

3.6.2. Conducting a test

A portion of peptone weighing 1.0-1.5 g, weighed with an error of not more than 0.0002 g, is placed in a crucible, previously calcined to constant weight. 1 cm3 of concentrated sulfuric acid is added to the crucible with a peptone weight and the mixture is carefully heated on a hot plate until excess sulfuric acid is removed, after which the crucible is transferred to a muffle furnace and ashing is continued at a temperature of 500-600 ° C to a constant mass of ash.

3.6.3. Results processing

The mass fraction of sulfonated ash () as a percentage is calculated by the formula

where is the mass of peptone before calcination, g;

- mass of ash after calcination, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

(Changed edition, Rev. N 2).

3.7. Determination of the mass fraction of true peptone

3.7.1. Equipment and reagents

For testing use:

photoelectric colorimeter or spectrophotometer;

desktop centrifuges type OPn-8-U4,2;

pipettes with a capacity of 1 and 5 cm according to GOST 29169-91;

glass test tubes according to GOST 25336-82;

sodium oxide hydrate according to GOST 4328-77, 10% solution;

copper sulfate according to GOST 4165-78, 2% solution;

distilled water according to GOST 6709-72.

(Changed edition, Rev. N 2).

3.7.2. Preparing for the test

Construction of a calibration graph

The abscissa shows the concentration of true peptone in the solution, the ordinate shows the value of the optical density () of this solution according to the data given in Table 2. The values ​​of optical density () given in Table 2 were obtained by measuring on a photoelectric colorimeter at a wavelength of 540 nm solutions of a specially prepared series of peptone containing a minimum of free protein and amino acids and a maximum of peptides.

table 2

3.7.3. Conducting a test

True peptone is determined in a 0.5% solution of the tested peptone. To do this, a 5% peptone solution prepared in accordance with clause 3.4.2 is diluted 10 times with water (0.5 ml of a 5% peptone solution and 4.5 cm3 of distilled water).

In a test tube containing 5 cm3 of a 0.5% solution of the test peptone, add 0.5 cm3 of a 10% solution of sodium hydroxide and 0.5 cm3 of a 2% solution of copper sulphate. At the same time, a control is placed, for which 5 cm3 of water, 0.5 cm3 of a 10% solution of sodium hydroxide and 0.5 cm3 of a 2% solution of copper sulphate are poured into a test tube.

The mixture in test tubes is well mixed and centrifuged for 10 minutes at a rotation frequency of 5000-6000 minutes. The color intensity is measured on a photoelectrocolorimeter at a wavelength of 540-560 nm or on a spectrophotometer at a wavelength of 540 nm in cuvettes with a working length of 10 mm. The concentration of true peptone in the sample is determined by the calibration curve.

(Changed edition, Rev. N 2

3.7.4. Results processing

The mass fraction of true peptone () as a percentage is calculated by the formula

where is the content of true peptone in the test solution, determined according to the calibration curve,%;

- mass of peptone (preparation) contained in 100 g of a 0.5% solution, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 5%.

(Changed edition, Rev. N 2).

3.8. Determination of the mass fraction of total nitrogen

(Changed edition, Rev. N 2).

The content of total nitrogen is determined by the Kjeldahl method or by the Kjeldahl method in combination with isothermal distillation in Conway cups.

3.8.1. Determination of the mass fraction of total nitrogen by the Kjeldahl method

(Changed edition, Rev. N 2).

3.8.1.1. Equipment and reagents

For testing use:

analytical balances;

GOST 25336-82;

apparatus for stripping ammonia;

pipettes with a capacity of 1, 2, 5 cm in accordance with GOST 29169-91;

burettes with a capacity of 25 or 50 cm3 with a division value of 0.05 cm3 in accordance with GOST 29252-91;

distilled water according to GOST 6709-72;

sulfuric acid according to GOST 4204-77, concentrated and 0.1 n. solution;

sodium oxide hydrate according to GOST 4328-77, 30-33% and 0.1 n. solutions prepared with pre-boiled distilled water;

hydrogen peroxide according to GOST 10929-76;

methyl red;

methyl blue;

GOST 5962-67 *;
________________
* In the territory Russian Federation valid GOST R 51652-2000. Here and further. - Database manufacturer's note.

methyl orange, 0.1% solution.

3.8.1.2. Conducting a test

1 ml of a 5% peptone solution is placed in a Kjeldahl flask and 2 ml of concentrated sulfuric acid are added. A glass pear-shaped canister or a small funnel with a sealed tip is inserted into the neck of the flask and the sample is first mineralized over low heat, then the degree of heating is increased, preventing the boiling mixture from being thrown out into the narrow part of the flask. Mineralization is carried out in the presence of a hydrogen peroxide catalyst. The catalyst is added after the start of boiling by 0.5 cm every 15-20 minutes until the solution is completely discolored.

After cooling, the contents of the flask are transferred from 10-12 ml of distilled water to a distillation flask. The completeness of the transfer is checked by the methyl orange indicator. The last portion of flushing water should remain yellow.

In the receiving flask of the Kjeldahl apparatus, pour 20 ml of 0.1 N. sulfuric acid solution and 2 drops of Tashiro indicator. The Tashiro indicator is prepared by dissolving 0.4 g of methyl red and 0.2 g of methyl blue in 200 ml of 96% ethanol.

The stripping flask is connected to a condenser and a vaporizer. The contents of the stripping flask are neutralized by methyl orange indicator with 30-33% sodium hydroxide solution. The distillation is carried out until about 70 cm3 of solution is collected in the receiving flask.

The contents of the receiving flask are titrated with 0.1 N. sodium hydroxide solution until the color of the solution changes from purple to green. At the same time, they put a control experiment on reagents, for which, instead of a peptone solution, 1 cm3 of distilled water is taken into the flask for mineralization

3.8.1.3. Results processing

where is the amount of 0.1 n. sulfuric acid solution poured into a receiving flask, cm;

- correction factor to the titer 0.1 n. sulfuric acid;

- the amount of 0.1 n. sodium hydroxide solution used for titration of the test or control sample, cm;


- mass of peptone contained in 1 cm 5% solution taken for analysis, g;

- the amount of nitrogen equivalent to 1 cm 0.1 N. sulfuric acid solution,

The final test result is the difference between the arithmetic mean of the results of two parallel determinations in experimental and control samples. Permissible discrepancies between the results of two parallel determinations should not exceed 0.2%.

(Revised edition, Rev. N

3.8.2. Determination of the mass fraction of total nitrogen by the Kjeldahl method in combination with isothermal distillation in Conway dishes

(Changed edition, Rev. N 2).

3.8.2.1 Apparatus and reagents

For testing use:

microburettes;

pipettes with a capacity of 1, 2, 5, 10 cm in accordance with GOST 29169-91;

Conway cups;

Kjeldal flasks with a capacity of 50-100 cm 3 according to GOST 25336-82;

volumetric flasks with a capacity of 100 cm3 in accordance with GOST 1770-74;

vaseline according to GOST 5774-76;

hydrogen peroxide according to GOST 10929-76;

methyl red;

methylene blue;

ethyl alcohol rectified according to GOST 5962-67;

sulfuric acid GOST 4204-77, concentrated and 0.025 n. solution;

sodium oxide hydrate (caustic soda) according to GOST 4328-77, 40% and 0.025 n. solutions;

distilled water according to GOST 6709-72.

3.8.2.2. Preparing for the test

Preparation of the Tashiro indicator according to clause 3.8.1.2.

3.8.2.3. Conducting a test

5 ml of a 5% peptone solution are mineralized with heating in a Kjeldahl flask with 10 ml of sulfuric acid (concentrated) until a colorless solution is obtained. 0.5 cm of perhydrol is used as a catalyst. After mineralization, the sample was quantitatively transferred into a 100 ml volumetric flask and made up to the mark with distilled water. Pour exactly 2 cm of 0.025 N into the inside of the Conway dish. sulfuric acid solution. To prevent the liquid used from spreading, the cup is set obliquely. Then close the lid, smeared with petroleum jelly, leaving a small gap, through which 1 cm of the test solution after mineralization is introduced into the outer part of the cup with a pipette. The cup is moved so that a gap of 5-6 mm wide remains on the opposite side, and 4 cm of a 40% alkali solution is quickly poured through it. Close the cup tightly with a lid, carefully mix the contents in a circular motion and leave for 15-18 hours. sodium hydroxide solution according to the Tashiro indicator. In parallel, a control sample is placed, for which distilled water is placed in the outer part of the Conway cup instead of the test solution.

3.8.2.4. Results processing

The mass fraction of total nitrogen () as a percentage is calculated by the formula

where is the amount of 0.025 n. sodium hydroxide solution used for titration of the control sample, cm;

- the amount of 0.025 n. sodium hydroxide solution used for titration of the test sample, cm;

- correction factor to the titer 0.025 n. sodium hydroxide solution;

- the amount of nitrogen equivalent to 1 cm 0.025 N. sodium hydroxide solution, g;

- mass of peptone taken to obtain a 5% solution, g;

- the amount of the drug taken for mineralization, ml;

- the amount of the test solution, see

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

(Revised edition, Rev.

3.9. Determination of the mass fraction of nitrogen of amino groups of amino acids and lower peptides

(Changed edition, Rev. N 2).

3.9.1. Equipment and reagents

For testing use:

pH meter;

laboratory glasses according to GOST 25336-82;

pipettes with a capacity of 1, 2 and 20 cm in accordance with GOST 29169-91;

burettes with a capacity of 5 and 10 cm3 with a division value of 0.05 cm3 in accordance with GOST 29252-91;

formalin technical according to GOST 1625-89;

sodium oxide hydrate according to GOST 4328-77, 0.1 n. solution;

sulfuric acid according to GOST 4204-77, 0.1 g * solution;

phenolphthalein according to NTD, 1% alcohol solution;

distilled water according to GOST 6709-72.
________________

3.9.2. Preparing for the test

Immediately before work, the formol mixture is prepared. To do this, add 1 cm of a 1% solution of phenolphthalein to 50 cm of formalin and 0.1 g * of sodium hydroxide solution to bring the color of the mixture to slightly pink.
________________
* Corresponds to the original. - Database manufacturer's note.

3.9.3. Conducting a test

To 2 cm of a transparent 5% peptone solution prepared in accordance with clause 3.4.2, add 18 cm of distilled water and adjust the pH of the mixture to 7.0 potentiometrically 0.1 N. a solution of sodium hydroxide or sulfuric acid. Then 2 ml of the formol mixture is poured and, with constant stirring, titrated potentiometrically with 0.1 N. sodium hydroxide solution to pH 9.2.

At the same time, a control is placed on the reagents, for which, instead of 2 cm3 of 5% peptone, 2 cm3 of distilled water are taken.

3.9.4. Results processing

The mass fraction of nitrogen of amino groups of amino acids and lower peptides in peptone () as a percentage is calculated by the formula

where is the amount of 0.1 n. sodium hydroxide solution used for titration of the test sample, cm;

- the amount of 0.1 n. sodium hydroxide solution used for titration of the control sample, cm;

- correction factor to the titer 0.1 n. sodium hydroxide solution;

- the amount of nitrogen equivalent to 1 cm 0.1 N. sodium hydroxide solution, g;

- mass of peptone contained in 2 cm3 of 5% peptone, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

(Revised edition, Rev. N

3.10. Determination of free protein

(Changed edition, Rev. N 2).

3.10.1. Equipment and reagents

For testing use:

photoelectric colorimeter;

pipettes with a capacity of 5 and 10 cm in accordance with GOST 29169-91;

glass test tubes according to GOST 25336-82;

trichloroacetic acid 20% solution.

3.10.2. Conducting a test

To 5 cm3 of the 5% peptone solution filtrate prepared in accordance with clause 3.4.2, add 5 cm3 of a 20% trichloroacetic acid solution and mix well. After 5 minutes, the optical density of the mixture is determined on a photoelectric colorimeter at a wavelength of 630 nm (red light filter) in cuvettes with a working length of 10 mm against the same peptone solution diluted 1:1 with distilled water.

3.10.3. Results processing

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.05%.

3.11. Determination of the mass fraction of chlorides

(Changed edition, Rev. N 2).

3.11.1. Equipment and reagents

For testing use:



heating element (burner, stove);

pipettes with a capacity of 1, 2, 10, 20 and 25 cm in accordance with GOST 29169-91;

burettes with a capacity of 25 cm3 with a division value of 0.05 cm3 in accordance with GOST 29252-91;

laboratory glasses according to GOST 25336-82;

distilled water according to GOST 6709-72;

silver nitrate according to GOST 1277-75, 0.1 n. solution;

ferroammonium alum according to NTD, saturated solution;

potassium thiocyanate according to NTD or ammonium thiocyanate 0.1 n. solution;

nitric acid according to GOST 4461-77, concentrated.

(Changed edition. Rev. N 2).

3.11.2. Conducting a test

To 20 ml of a 1% peptone solution prepared in accordance with clause 3.4.2, add 30 ml of distilled water, 1 ml of concentrated nitric acid and 10 ml of 0.1 N. silver nitrate solution. The solution is heated to boiling and quickly cooled by immersing the beaker of the solution in a vessel of water at room temperature. After cooling the mixture, add 2 ml of a saturated solution of iron ammonium alum (indicator) and titrate with 0.1 N. a solution of potassium thiocyanate (or ammonium thiocyanate) until a brick-red color appears.

3.11.3. Results processing

The mass fraction of chlorides () as a percentage is calculated by the formula

where is the amount of 0.1 n. a solution of silver nitrate, taken into a beaker with a sample, cm;

- correction factor to the titer 0.1 n. silver nitrate solution;

- the amount of 0.1 n. solution of potassium thiocyanate or ammonium thiocyanate used for titration of the sample, cm;

- correction factor to the titer 0.1 n. a solution of potassium thiocyanate or ammonium thiocyanate;

- the amount of chlorides (chlorine ion), equivalent to 1 ml of 0.1 N. silver nitrate solution, g;

- the mass of peptone contained in 20 cm3 of a 1% solution, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.1%.

(Revised edition, Rev. N

3.12. Determination of salts of heavy metals

3.12.1. Equipment and reagents

For testing use:

technical scales with a weighing error of not more than 0.01 g;

heating element (stove, burner); water distilled with concentrated sulfuric acid and carefully ashed first on a stove or burner, and then in a muffle furnace at 500-600 ° C until a homogeneous light gray ash is obtained.

After cooling the crucible, the ash is poured into 1 cm3 of a saturated solution of ammonium acetate and diluted with 5 cm3 of distilled water. The contents of the crucible are filtered into the cylinder through a paper filter. The crucible and filter are washed with distilled water, bringing the volume of the solution in the cylinder to 10 cm3.

The reaction of the filtrate in the cylinder is brought to neutral or slightly acidic by litmus by adding dropwise 10% acetic acid or 10% sodium hydroxide solution. Then add 1 ml of 10% acetic acid, 5 drops of a saturated solution of sodium sulfide and mix. After 5 minutes, the solution should not have a precipitate and a yellow color of the solution.

3.13. Determination of the presence of free indole

3.13.1. Equipment and reagents

For the test, apply: 0.1% solution;

sulfuric acid according to GOST 4202-75 *, concentrated.
_________________
* Reference to the standard corresponds to the original. - Database manufacturer's note.

3.13.2. Conducting a test

Add 2 ml of amyl alcohol to a test tube with 10 cm3 of 1% peptone solution, close the tube with a cork, shake well, and add 1 cm3 of 0.1% potassium nitrite solution and 1 cm3 of concentrated sulfuric acid. The mixture is gently stirred and allowed to settle.

No pink coloration should appear in the alcohol layer. The appearance of a pink coloration of the alcohol layer indicates the presence of free indole in the peptone.

3.14. Determination of the ability to support microbial growth

3.14.1. Equipment, materials and reagents

For testing use:

technical scales with a weighing error of not more than 0.01 g;

pH meter;

thermostat at 37 °C;

autoclave;

photoelectric colorimeter;

test tubes according to GOST 25336-82;

cotton-gauze corks;

micropipettes with a capacity of 0.2 cm;
nutrient media (meat water according to
To prepare MPA, 1% of the test peptone, 0.5% sodium chloride, 2% microbiological agar are added to the meat water and the pH of the medium is set to 7.4.

MPB and MPA are filtered, poured into 8 cm test tubes, closed with cotton-gauze stoppers and parchment caps and sterilized in an autoclave for 30 minutes at 0.15 MPA.

3.14.3. Conducting a test

Test tubes with sterile BCH and oblique MPA are inoculated with daily control cultures. Each culture is inoculated with a 0.2 cm micropipette into three test tubes with MPB and MPA. Incubation is carried out for 24 hours at 37-38 °C.

The typical growth of daily cultures is determined on the MPA visually and under a microscope. Growth should be typical of each strain* of microorganisms. The growth intensity is evaluated in the BCH on a photoelectrocolorimeter at a wavelength of 620-640 nm in a cuvette with a working length of 5 mm.
________________
* The text corresponds to the original. - Database manufacturer's note.

4. PACKAGING, PACKAGING, MARKING, TRANSPORTATION AND STORAGE

4.1. Peptone is packaged with a net weight of 250 to 1000 g in cans according to GOST 5981-88 or, by agreement with the consumer, with a net weight of 30 kg in type II plywood barrels according to NTD with a polyethylene film lining in accordance with GOST 10354-82. The cans are hermetically sealed.

(Changed edition, Rev. N 1).

4.2. The jar is labeled with:

name of the manufacturer;

name of the drug;

series numbers;

net weight;

storage conditions;

expiration date;
the name of the manufacturer and his trademark;

name of the drug;

series numbers;

manufacturing dates;

gross and net weights;

storage conditions;

expiration date;

designations of this standard.

A handling sign "Afraid of dampness" is applied to each box or barrel.

(Changed edition, Rev. N 1).

4.5. Peptone is stored in a dry room at a temperature not exceeding 30 °C. The shelf life of peptone in hermetically sealed jars is 3 years, in plywood barrels - 1 year from the date of manufacture.

4.6. Peptone is transported by all means of transport in compliance with the rules for the carriage of goods in force on this type of transport.

Strengthening of packages in overpacks when transporting goods by rail in accordance with GOST 24597-81 and GOST 26663-85.

(Changed edition, Rev. N 1).

Electronic text of the document
prepared by Kodeks JSC and verified against:
official publication
M.: IPK Standards Publishing House, 1995

UDC t.ShZ-.SHMA Pear "H18 STATE STANDARD OF THE UNION SSR PEPTONE DRY ENZYMATIVE FOR BACTERIOLOGICAL PURPOSES Te> nmch * sima condition E (u Gsgtel" aKop rsr (op! 01-M Lastaioayaenom Go * "door of the secret committee * of the standards of the Council Minister of O * USSR in * 3 * mrem IN g. and * IP g. obtained from the rumen and lstochka of cattle, sheep and cola, as well as from the stomachs of pigs using the mucous membrane of the stomach and pancreas, 1, TECHNICAL REQUIREMENTS 1.1, Enzymatic peptone for bacteriological purposes must be manufactured in accordance with the requirements of this standard according to the technological rules 2. Enzymatic peptone for bacteriological purposes must comply with the requirements and rmam specified in table. I. Table! X"f>:m(f"!-™»> K H1fK1 External vn.ch Color Zavakh Amorphous oysteric powder From Gimoy to yellow Characteristic, without putrefaction Official publication Reprint prohibited Reprint April 1980

Page 2 GOST "*0> -76 Continued gay.!. I I 1.11-1 |||-.»_n II- 1ISHL1SLP Concentrations of nolyride rest (OH) Content of insoluble impurities. %. no more than Content of magician, %, no more no more Content of true peptone. %, not less than Content of the total act, %, not less than Content of amino groups of amino groups and niches of peptides, %, not less Free protein content, optical density, not more than Content of chlorides in terms of hyaor-non. ve Solee Content of salts of heavy metals Presence of free indole Ability to support the growth of microbes, optical density, "not less than 5 * arnucococi" jasgei * "Lassmanov" E&spepsia coli strain 675 51 * eplococci* laesa !!& strain 6783 at "C<л-7,0 1,0 7.0 5.0 70,0 1-У) а* Не допускается То же Рост типичный 0.40 03* О.» 2.1. Ферментативный пептон для бактериологических целей принимают сериями. Под серией следует понимать количество пептона, изготовленное из одноименного вида сырья за один технологический цикл, получившее свой номер и оформленное одним документом о качестве. 2.2. Дли контроля качества ферментативного пептона от каждой серии препарата отбирают выборку в размере: от серии до 104) единиц фасовки - 3%; от серн» свыше 100 единиц фасовки - 1%. 2.3. Выборку отбирают из единиц фасовки, взятых из разных мест серии. 2.4. При неудовлетворительных, результатах испытаний хотя бы по одному показателю по нему проводят повторные испытания на удвоенном количестве проб, отобранных от тон же серии пептона. Результаты испытаний распространяются на всю серию. 3. МЕТОДЫ ИСПЫТАНИЙ 3.1. Метод отбора проб 3.1.1. Содержимое отобранных единиц фасовки тщательно перемешивают н отбирают общую пробу массой не более 500 г.

GOST 11801-76 Or. 3 Four coffins for analysis weighing at least 100 g each are isolated from the total sample of enzymatic peptone. The selected samples will be placed in clean, dry jars with ground-in stoppers. Corks are filled with paraffin or resin. 3.12. On jars with samples of enzymatic peptone stick labels indicating: the name of the manufacturer; name of the drug; manufacturing dates; series numbers; date of sampling; designations of this standard. Two samples are used for analysis, and two are left in the archives of the State Comptroller for 4 years. 3 2. External fork. the color and smell of peptone is determined organoleptically. 3.3. Definition of the concept of hydrogen ions (pH) 3.3.1. Apparatus and reagents For testing use; pH meter; (laboratory scales; * glass flask according to GOST 1770-74; glass measuring cylinder with a capacity of 100 ml according to GOST 1770-74; paper filters: distilled water according to GOST 6709-72. 3.3.2. Testing with an error of not more than 0.02 g, placed in a flask and dissolved in 99 ml of boiled and cooled distilled water. A slight precipitate is allowed in the solution, which is filtered through a paper filter. In a transparent filtrate of a 1% peptone solution, the concentration of hydrogen ions (pH) is determined in accordance with 3.4.Determination of the content of insoluble impurities 3.4.1.Apparatus, materials and reagents For testing, analytical balances, drying oven at 105°C, weighing cups (bottles) according to GOST 7148-70 are used. ; desiccator in accordance with GOST 6371-73; glass measuring cylinder with a capacity of 100 ml in accordance with GOST 1770-74; cork cotton-gauze;

Page 4 GOST 13805^-7* paper filters; sodium oxide hydrate according to GOST 4328-77. 0.1 i. solution, distilled water according to GOST 6709-72. 3.4.2. Testing A peptone weighing 5 g, weighed with an error of more than * 0.0002 g, is placed in a flask, 95 ml of distilled water are added and thoroughly mixed, achieving maximum peptone dissolution. The prepared 5% solution of nepton is filtered through a paper filter, preliminarily dried in a weighing bottle to constant weight and weighed with an error of not more than 0.0002 g. After filtering the peptone solution, the filter is washed with 50 ml of distilled water, placed in a weighing bottle and dried to constant weight at! 05 * C. The first weighing is carried out after drying the filter for 3 hours. Subsequent weighings - after 30 minutes of drying. Drying is considered complete if the decrease in mass during two subsequent weighings does not exceed 0.0002 g. The filtered solution of 5% peptone is diluted 5 rad with distilled water. The resulting 1% peptone solution is adjusted with 0.1 k sodium hydroxide solution to pH 7.7 and boiled for 5 minutes. The solution should remain clear, without the formation of a precipitate. After boiling, the 1% solution of peptone is poured into test tubes of 7-8 ml. close with cotton plugs and incubated in an autoclave for 30 min prn 0.15 MPa. The solution should remain clear, white precipitate formed. 3.4.3. Processing of results The content of insoluble impurities (X) in percent is calculated by the formula d-- (Dg-ln) -400 where m is the mass of peptone taken to prepare 100 g of a 5% peptone solution, g; t, is the weight of the weighing bottle with the filter before filtration, g; m3 is the mass of the weighing bottle and the filter with sediment after drying, g. The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel surveys should not exceed 0.1 V 35 Determination of moisture content 3.5.1, Apparatus For the test, the following are used: analytical balance;

GOST L Page 5 laboratory drying cabinet at 105°С; cups for weighing (bottle bags) according to GOST 7148-70; desiccator according to GOST 6371-73. 3.5.2. Testing A peptone weighing 1.5-2.0 g, weighed with an error of not more than 0.0002 g, is placed in a weighing bottle, previously dried to a constant weight. The weighed bottle is placed in an oven and dried to constant weight at 105°C. The first weighing is carried out after 1 hour, the next - after 30 minutes of drying. Drying is considered complete if the decrease in weight during two subsequent weighings does not exceed 0.0002 g. 3.5.3. Processing of results Moisture content (X,) in percent is calculated by the formula t, is the weight of the peptone sample after drying, g. The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%. 3.6. Determination of the content of sulfonated ash 3.6.1. Apparatus and reagents For testing, the following are used: analytical weights; heating element (tile, burner); muffle furnace; porcelain crucibles according to GOST 9147-73; sulfuric acid according to GOST 4204-77, concentrated. 3.6.2. Testing A weighed portion of peptone weighing 1.0-1.5 g, weighed with an error of not more than 0.0002 g, is placed in a crucible, previously calcined to constant weight. 1 ml of concentrated sulfuric acid is added to a crucible with a peptone weight and the mixture is carefully heated on a hot plate until excess sulfuric acid is removed, after which the crucible is transferred to a muffle furnace and ashing is continued at a temperature of 500-600 * C to a constant mass of ash. 3.6.3. Processing of results The content of sulfonated ash (Xg) as a percentage is calculated by the formula V "I *" 100 where /l is the mass of peptone before calcination, g;

Page 6 GOSG 11801-76 The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%. 37. Determination of the content of true peptone 3.7.1. Equipment and reagents For testing use: photoelectric colormeter or spectrophotometer; pipettes with a capacity of I n 5 ml; glass test tubes according to GOST 10515-75; glass funnels; paper filters; sodium oxide hydrate according to GOST 4328-77, 10% solution; copper sulfate according to GOST 4165-78, 2% solution; distilled water according to GOST 6709-72. 3.7.2. Preparation for the test Construction of a calibration graph The concentration of true peptone in the solution is plotted along the abscissa axis, the value of the optical density (E) of this solution is plotted along the ordinate axis according to the data given in Table. 2. Given in table. 2 values ​​of optical density (E) were obtained by measuring solutions of a specially prepared series of peptone containing a minimum of free protein and amino acids and a maximum of peptides on a photoelectrocolormeter at a wavelength of 540 nm. Table 2 K "aa

GOS1 PVO!-7* Page 7 0.5 ml of 10% sodium hydroxide solution and 0.5 ml of 2% sulfuric acid solution. Mix well in test tubes and after 2 minutes filter through paper filters. The color intensity is measured on a photomectrocolorimeter at a wavelength of 540-560 nm or on a spectrophotometer at a wavelength of 540 nm in cuvettes with a working length of 10 mm. The concentration of true peptone in the sample is determined by the calibration curve. 3.7.4. Processing of results The content of true peptone (X ") in percent is calculated using the formula y A pum where A is the content of true peptone in the test solution, determined according to the calibration graph. %; 0.5-mass of peptone (preparation) contained in 100 g of a 0.5% solution, g. The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 5%. 3.8. Determination of total nitrogen content The total nitrogen content is determined by the Kjeldahl method or by the Kjeldahl method and combined with isothermal distillation in Conway dishes. 3.8.1. Determination of the content of total nitrogen by the Kyell-lal method 3.8.1.1. Apparatus and reagents For testing use: analytical balance; Kjeldal flasks with a capacity of 50-100 ml according to GOST 10394-72; apparatus for stripping ammonia; pipettes with a capacity of 1, 2. 5 ml:. burettes with a capacity of 25 or 50 ml with a division value of 0.05 ml; distilled water according to GOST 6709-72; sulfuric acid according to GOST 4204-77. concentrated n 0.1 in. solution; sodium hydrate oxide but GOST 4328-77. 30-33% and 0.1 n. solutions prepared with pre-boiled distilled water; hydrogen peroxide according to GOST 10929-76; methyl red according to GOST 5853-51; methyl blue; ethyl alcohol rectified according to GOST 5962-67;

Sgr. In GOST 1Sh5-76 methyl orange according to GOST 10816-64. 0.1% kP solution. 3.8.1.2. Conducting the test In a Kjeldahl flask, place 1 ml of a 5% peptone solution and add 2 ml of concentrated sulfuric acid. A glass pear-shaped canister or a small funnel with a sealed tip is inserted into the neck of the flask and the sample is first mineralized over low heat, then the degree of heating is increased, preventing the boiling smog from being thrown into the narrow part of the flask. Mineralization is carried out in the presence of a hydrogen peroxide catalyst. The catalyst is added after the start of boiling in 0.5 ml increments every 15-20 min until the solution becomes completely colorless. After cooling, the contents of the flask are transferred from 10-12 ml of distilled water to a flask for distillation. The completeness of the transfer is checked by the methyl orange indicator. The last portion of flushing water should remain yellow. In the receiving flask of the Kjeldahl apparatus, pour 20 ml of 0.1 N. sulfuric acid solution and 2 drops of Tashiro indicator. The Tashro indicator is prepared by dissolving 0.4 g of methyl red in 0.2 g of methyl blue in 200 ml of 96% ethanol. The stripping flask is connected to a condenser and steam generator. The contents of the stripping flask are neutralized by methyl orange indicator with 30-33% sodium hydroxide solution. Distillation is carried out until then. until about 70 ml of solution is collected in the receiving flask. The contents of the receiving flask are titrated with 0.1 i. sodium hydroxide solution until the color of the solution changes from purple to green. At the same time, a control experiment is put on the reagents, for which, instead of a peptone solution, a flask for mineralization is taken with 1 ml of distilled water. 3.8.1.3. Processing of results The content of total nitrogen (N *.) as a percentage is calculated by the formula x, - - ^_-. amount 0.1 n. sulfuric acid solution poured into a receiving flask, ml; correction factor to the titer 0.1 n. sulfuric acid; amount 0.1 n. sodium hydroxide solution used for titration of the tested nln control sample, ml; correction factor to the titer 0.1 n. a solution of a pirate window and aatria; mass of peptone contained in 1 ml of a 5V solution, i 1 for analysis, g; where V- / C - V, - K, -0.05 -

GOST NII-74 Page 9 0.0014 - the amount of nitrogen equivalent to I ml of 0.1 n. sulfuric acid solution, g. The difference between the arithmetic mean of the results of two parallel determinations in experimental and control samples is taken as the final test result. Permissible discrepancies between the results of two parallel determinations should not exceed 0.2%. 3.8.2. Determining the content of total nitrogen by the Kjeldahl method in combination with isothermal distillation in Conway dishes 3.8.2.1. Apparatus and reagents For testing use; microburettes; pnpetkn with a capacity of I, 2, 5, 10 ml; Conway cups; Kjeldal flasks with a capacity of 50-100 ml according to GOST 10394-72; volumetric flasks with a capacity of 100 ml according to GOST 1770-74; vaseline according to GOST 5774-76; hydrogen peroxide according to GOST 10929-76; methyl red according to GOST 5853-51; methylene blue; ethyl alcohol rectified according to GOST 5962-67; sulfuric acid GOST 4204-77, concentrated acid and 0.025 n. solution; sodium oxide hydrate (caustic soda) according to GOST 4328-77, 40% and 0.025 c. solutions; distilled water according to GOST 6709-72. 3.8.2.2. Preparation for the test Preparation of the Tashiro indicator according to clause 3.8.1.2. 3.8.2.3. Conducting the test 5 ml of a 5% peptone solution is mineralized with heating in a Kjeldahl flask with 10 ml of sulfuric acid (concentrated) ♦ until a colorless solution is obtained. 0.5 ml of perhydrol is used as a catalyst. After mineralization, the sample was quantitatively transferred into a 100 ml volumetric flask and made up to the mark with distilled water. Pour exactly 2 ml of 0.025 i into the inside of the Conway cup. sulfuric acid solution. To prevent the liquid used from spreading, the cup is set obliquely. Then close the lid, smeared with vaseline, leaving a small opening, through which 1 ml of the test solution after mineralization is pipetted into the outer part of the cup. The cup is moved so that a 5-6 mm wide slit remains on the opposite side, and 4 ml of a 40% alkali solution is quickly poured through it. Close the cup tightly with a lid, gently mix the contents in a circular motion and leave for 15-18 hours.

Page 10 GOST TsVM-76 titrating 0.025 N. sodium hydroxide solution according to the Tashiro indicator. At the same time, a control sample is placed, for which distilled water is placed in the outer part of the Convoy cup instead of the test solution. 3.8.2.4. Processing of results The content of total nitrogen (X $) in percent is calculated by the formula “0.00035 (1 "- Y,) K-Yu0.1OO-1OO 6-5-1 where K is the amount of 0.025 N. sodium hydroxide solution consumed on titration of the control sample, ml V. is the amount of 0.025 N sodium hydroxide solution used for titration of the test sample, ml K is the correction factor for the titer of 0.025 N sodium hydroxide solution 0.00035 is the amount of nitrogen equivalent to 1 ml of 0.025 N sodium hydroxide solution caustic soda, g; 5-mass of peptone taken to obtain a 5% solution, g; 5-amount of the drug taken for mineralization, ml;] - the amount of the test solution, ml. For the final test result, take the arithmetic mean of the results of two parallel 3.9.Determination of nitrogen content of amino groups of amino acids and lower peptides 3.9.1.Apparatus and reagents according to GOST 10394-72; pipettes with a capacity of 1, 2 and 20 ml; burettes with a capacity of 5 and 10 ml with a division value of 0.05 ml; formalin technical according to GOST 1625-75; sodium oxide hydrate according to GOST 4328-77, 0.1 n. solution; sulfuric acid according to GOST 4204-77, 0.1 n. solution; phenolphthalein according to GOST 5850-72, 1% spring solution; distilled water according to GOST 6709-72. 3.9.2. Preparation for the test Immediately before work prepare the formol mixture. To do this, 1 ml of a 1% solution of phenolphthalein and 0.1 i.m. are added to 50 ml of formalin. With sodium hydroxide solution, the color of the mixture is brought to slightly pink.

GOST 13805-74 Page 11 3.9.3. Carrying out the test To 2 ml of a clear 5% peptone solution, prepared in accordance with paragraph 3.4.2. 18 ml of distilled water are poured and the pH of the mixture is adjusted to 7.0 by 0.1 n. sodium hydroxide solution or sulfuric acid solution. Then 2 ml of the formal mixture is added and, with constant stirring, titrated potentiometrically with 0.1 N sodium hydroxide. sodium hydroxide solution to pH 9.2. At the same time put the control on the reagents, for which instead of 2 ml of 5% peptone take 2 ml of distilled water. 3.9.4. Processing of results The content of nitrogen of amino groups of amino acids and lower peptides in peptone (X.) in percent is calculated by the formula * -^ "I-. where V is the amount of 0.1 n. sodium hydroxide solution used for titration of the test sample, ml; V)- quantity 0.1 and. sodium hydroxide solution used for titration of the control sample, ml; K - correction factor to the titer of 0.1 n. sodium hydroxide solution; 0.0014 - the amount of nitrogen equivalent to 1 or 0.1 k. of sodium hydroxide solution, g; 0.1 - peptone oils contained in 2 ml of 5% peptone, g. The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%. 3 10. Determination of free protein content 3 10.1. Apparatus and reagents For the test, the following are used: photo pipettes with a capacity of 5 and 10 ml; glass test tubes according to GOST 10515 75; trichloroacetic acid 20% solution. 3.10.2. Carrying out the test To 5 ml of the filtrate of a 5% peptone solution, prepared in accordance with paragraph 3.4.2. add 5 ml of a 20% trichloroacetic acid solution and mix well. After 5 minutes, the optical density of the mixture is determined on a photoelectric colorimeter at a wavelength of 630 nm (red light filter) in cuvettes with a working length of 10 mm against the same peptone solution diluted 1: I with distilled water. 3.10.3, Handling results

Page I? GOST 13105-76 The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.05%, 3.11. Determination of chloride content 3.11.1. Apparatus and reagents For the test, the following are used: technical scales with a weighing error of not more than 0.01 g; heating element (burner, stove); pipettes with a capacity of I, 2. 10, 20 I 25 silt; burettes with a capacity of 25 ml with a division value of 0.05 ml; laboratory glasses according to GOST 10394-72; distilled water according to GOST 6709-72; silver nitrate according to GOST 1277-75. 0.1 N solution; ferroammonium alum according to GOST 4205-77, saturated solution; potassium thiocyanate according to GOST 4239-77 or ammonium thiocyanate according to ST SEV 222-75, 0.1 i. solution; nitric acid according to GOST 4461-77, concentrated. 3.11.2. Carrying out the test To 20 ml of a 1% peptone solution prepared in accordance with paragraph 34 2. add 30 ml of distilled water, 1 ml of concentrated nitric acid and 10 ml of 0.1 liter. silver nitrate solution. The solution is heated to the beginning of the boil and quickly cooled by immersing the glass with the solution in a vessel with water at room temperature. After cooling the mixture, add 2 silt of a saturated solution of iron ammonium alum (indicator) and titrate with 0.1 K. a solution of potassium thiocyanate (or ammonium thiocyanate) until a reddish color appears. 3.11.3. Processing of results The content of chlorides "A".-) in percent is calculated according to the formula

goer ash-n Cgr, 13 The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.1% 3.12. Determination of salts of heavy metals 3.12.1. Apparatus and reagents For the test, the following are used: technical scales with a weighing error of not more than 0.01 g; heating element (tile, burner): muffle furnace; porcelain tngles according to GOST 9147-73; measuring cylinders with a capacity of 10 ml in accordance with GOST 1770-74; sulfuric acid according to GOST 4204-77. concentrated; ammonium acetate according to GOST 3117-78, saturated solution; distilled water according to GOST 6709-72; acetic acid according to GOST 61-75. 10% solution; sodium sulfide according to GOST 2053-77. saturated solution; sodium oxide hydrate according to GOST 4328-77, 10% solution; litmus or universal indicator. 3.12.2. Testing A weighed portion of peptone weighing 1 g, weighed with an error of not more than 0.02 g, is placed in a crucible, moistened with 2 ml of concentrated sulfuric acid and carefully ashed first on a stove or burner, and then in a muffle furnace at 500-600 "C until a homogeneous ash is obtained light gray color.After cooling the crucible, the ash is poured with 1 ml of a saturated solution of ammonium acetate and diluted with 5 ml of distilled water.The contents of the crucible are filtered into the cylinder through a paper filter.The crucible and filter are washed with distilled water, bringing the volume of the solution in the cylinder to 10 ml.Reaction of the filtrate in a cylinder, bring the litmus to neutral or slightly acid by adding dropwise 10% acetic acid or 10% sodium hydroxide solution, then add 1 ml of 10% acetic acid, 5 drops of saturated sodium sulphide solution and mix. 5 min in the solution there should be no precipitate and no yellow coloring of the solution 3.13 Determination of the presence of free nnolol 3.13.1 Apparatus and reagents assets are used for testing; glass test tubes according to GOST 10515-75; pipettes with a capacity of I. 2 and 10 silt; rubber cone plugs according to GOST 7852-76;

Page 14 GOST 13 * 05-7 * Add 2 ml of amyl alcohol to a test tube with 10 ml of a 1% solution of peptone, close the test tube with a cork, shake well, and add 1 ml of a 0.1% solution of potassium nitrate and 1 ml of concentrated sulfuric acid. The mixture is gently stirred and allowed to settle. No pink coloration should appear in the alcohol layer. The appearance of a pink coloration of the alcohol layer indicates the presence of free indole in the peptone. 3.14. Determination of the ability to support microbial growth 3.14.1. Equipment, materials and reagents For testing, the following are used: technical scales with a weighing error of not more than 0.01 g; pH meter; "thermostat at 37 ° C; autoclave; photoelectric color meter; test tubes according to GOST 10515-75; cotton-gauze stoppers; micropipettes with a capacity of 0.2 ml; pipettes with a capacity of 10 ml; burettes with a capacity of 25 or 50 ml, paper filters, nutrient media (meat water according to GOST 20729-75), microbiological agar according to GOST 17206-71 or other microbiological agar, sodium chloride according to GOST 4233-77. ) in accordance with the requirements of GOST 20730-75 and meat-peptone agar (MPA). To prepare MPB, add 1% of the test peptone to meat water. 0.5% sodium chloride and set the pH of the medium to 7.4. To prepare MPA, add 1% of the test to meat water peptone 0.5% sodium chloride, 2% microbiological agar and set the pH of the medium to 7.4.MPB and MPA are filtered, poured into test tubes in 8 ml, closed with cotton marl stoppers and parchment caps and sterilized in an autoclave for 30 minutes at 0.15 MPa. 3.14.3 Testing Test tubes with sterile MPB and oblique MPA are inoculated with daily control cultures. Inoculation of each culture is carried out with a micropipette of 0.2 ml in three test tubes with MPB and MPA. amyl alcohol; potassium aerosol according to GOST 4144-79, 0.1") o-th solution; sulfuric acid according to GOST 4202--75, concentrated.

GOST 11805-Te Or. 15 Incubation is carried out for 24 hours at 37-38*C. The typical growth of daily cultures is determined on MP A visually and under a microscope. Growth should be typical for each microorganism stamp. The intensity of growth is estimated in BCH on a photoelectric color meter at a wavelength of 620-640 nm in a cuvette with a working length of 5 mm. 4. PACKING, PACKAGING, MARKING, TRANSPORTATION AND STORAGE 4.1. Peptone is packaged with a net weight of 250 to 1000 g in cans in accordance with GOST 5981-71 or, by agreement with the consumer, with a net weight of 30 kg in plywood barrels in accordance with GOST 5958-79 with a polyethylene film lining in accordance with GOST 10354-73. The cans are hermetically sealed. 4.2. A label is stuck on the jar indicating: the name of the manufacturer: the name of the drug: the series number; net weight; storage conditions; expiration date; designations of this standard. 4.3. Banks with peptone are packed in plank non-separable boxes for canned food "in accordance with GOST 13358-72 or in corrugated cardboard boxes for canned food and food liquids in accordance with GOST 13516-72. 4.4. Each box and barrel is labeled in accordance with GOST 14192-77 with additional data: name of the manufacturer; name of the product; batch number; date of manufacture; gross and net weight; storage conditions; expiration date; designation of this standard. A warning sign is applied to each box or barrel. temperature not higher than 30"C. The shelf life of peptone in hermetically sealed jars is 3 years, in plywood barrels - 1 year from the date of manufacture. 4.6. Peptone is transported by all modes of transport. 4.7. In case of violation of the sealing of the container, peptone is stored in a tightly closed vessel or plastic bag in a dry room at a temperature of 0 to 8 ° C for no more than 1 month.

Ivmeienyas "" ■ GOST 13805-76 Dry enzymatic peptone, long o. mgrio-■from■-. .-li■ goals. Specifications By the Decree of the State Committee "of the USSR gn> standard *" t 31.1281 L 5930, the term Ivan is set from (11.01.82 Under the name of the standard, put the code: OKP 93 -481. Throughout the text of the standard, the unit of measurement will be measured: ml per cm1. Paragraphs 2.2, 23. 3.1.1. Replace the words; "single packing" with "cans". Paragraph 22 shall be filled with the words: "from serving in barrels - each barrel." Paragraph 2.3. Replace the words: "take away* with" make up. Replace the words: “from the same batch of peptone* with “from the same batch of samples*.” Paragraph 3.1.1, add the words: “When sampling* from barrels, peptone with a mass of at least 500 g is taken. Sample isolation for analysis is carried out as described above * Clauses 3.4.1, 3.7.1, 3.3.1.1, 3.8.2.1, 3.9.1, 3.12.1 Replace CN No. GOST 4328-66 with GOST 4328-77, GOST 2053-66 with GOST 2053-77. Clause 3.13.1. Replace references: GOST 7852-65 to GOST 7852-76. GOST 4144-65 to GOST 4144-7", GOST 4202-66 and GOST 4202-75. Item Z.I.1. Replace reference: GOST 4233-66 to GOST 4233-7?. Clause 4.1 after doe “plywood barrels? add the words: “type II”; See link: GOST 5958-70 pa GOST 5958-79. Clause 4.4. Replace the words: "Each box and barrel is marked in accordance with GOST 14192-71 with additional data * for "Transport marking in accordance with GOST 14192-77 with additional data", trademark"; "warning" to "mchnkpulyatsionny". Paragraph 4.6 shall be supplemented with the words: "in compliance with the rules for the carriage of goods in force on this type of transport. Strengthening of packages in transfer packages when transporting goods by the same * lemudorozhnich transport according to GOST 21929-76 and GOST 15901-TO*.

Group H18 I 1mg "ie" not l) 2 GOST 13805-Tv Peptone su * o * fsrmg11tl1i1 "th for baalernol * -g "P" (whole g l Technical conditions the deadline for ayedeya is set from aphrase Item 12. Table I Column "Name and" ii | geaya "" Replace the words "Free white content *, optical density, but more" with "Presence of free bsma, optical optical density, no more." "Content * heavy metal salts” to “Presence of heavy metal salts”; “Content” to “Mass fraction” (7 c) Paragraphs 3.4. 3.43. 35. 353. 3 6. 363. 3.7. 374, 38. 381. 13. 3*7. 88.24 39. 3.9.4. 3.11. 3 113 Replace "content" with "mass fraction" Clause 3.4 jealous *; (Continued from p. 290)

(Continuation of the changes in GOST 1Sh5-7b-> "3.71. Lpporaura and reagents For testing, they use: photomektrocolorkmeter or spechtrophoto: tabletop centrifuges OPK-8 N2, pnpegkn vmssgnmooyu I n 5 cm1 according to GOST glass test tubes according to GOST 25336-82 7. 10 % dilution; 2% solution: G 80292-74; sodium hydrate onisn according to I "l. I meya sulfate but GOST -I water dinititlyrvvimy according to GOST 6709-72". Item 373. After! a & aaa state in "o * oh relay; - The mixture in the prebcr-cad is well mixed and this rifugchruyug in "osava 10 willows * yari.iip. yarv-vaenaya 5000-6000 mim-" Nngensigmost coloring is iterated in the photomestrocolor-rameter arn at a wavelength of 540-560 im lv at snow trophometer at wavelength* 540 nor in cuvettes

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GOST 13805-76*

Group H18

STATE STANDARD OF THE UNION OF THE SSR

PEPTONE DRY ENZYMATIVE FOR BACTERIOLOGICAL PURPOSES

Specifications

Dry fermentation peptone for bacteriological purposes.
Specifications

Introduction date 1977-01-01

By the Decree of the State Committee of Standards of the Council of Ministers of the USSR dated April 29, 1976 N 1002, the introduction period was set from 01.01.77

The validity period was removed by decision of the Interstate Council for Standardization, Metrology and Certification (IUS 5-92)

INSTEAD OF GOST 13805-68

* REPUBLICATION (August 1995) with Amendments No. 1, 2, approved in December 1981, November 1986 (IUS 3-82, 2-87).


This standard applies to enzymatic peptone for bacteriological purposes, obtained from the rumen and lettuce of cattle, sheep and goats, as well as from the stomachs of pigs using the mucous membrane of the stomach and pancreas.

1. TECHNICAL REQUIREMENTS

1. TECHNICAL REQUIREMENTS

1.1. Enzymatic peptone for bacteriological purposes must be manufactured in accordance with the requirements of this standard according to the technological rules approved in the prescribed manner.

1.2. Enzymatic peptone for bacteriological purposes in terms of physicochemical and biological indicators must comply with the requirements and standards specified in Table 1.

Table 1

2. ACCEPTANCE RULES

2.1. Enzymatic peptone for bacteriological purposes is taken in series. A series should be understood as the amount of peptone made from the same type of raw material in one technological cycle, which has received its number and is issued with one quality document.

2.2. To control the quality of enzymatic peptone, a sample is taken from each batch of the drug in the amount of:

from a series to 100 cans - 3%;

from a series of more than 100 cans - 1%;

from a series in barrels - each barrel.

2.3. The sample is made up of jars taken from different places in the series.

2.4. In case of unsatisfactory test results for at least one indicator, repeated tests are carried out on it on a double number of samples taken from the same peptone sample.

The test results apply to the entire series.

2.2-2.4.

3. TEST METHODS

3.1. Sampling method

3.1.1. The contents of the selected jars are thoroughly mixed and a total sample is taken weighing no more than 500 g.

From the total sample of enzymatic peptone, four samples are isolated for analysis, each weighing at least 100 g. The selected samples are placed in clean, dry jars with ground stoppers. Corks are filled with paraffin or resin.

When sampling from barrels, peptone weighing 500 g is taken. Samples for analysis are isolated as described above.

(Changed edition, Rev. N 1).

3.1.2. Enzymatic peptone sample jars are labeled with:



name of the drug;

manufacturing dates;

series numbers;

date of sampling;



Two samples are used for analysis, and two are left in the archives of the State Comptroller for 4 years.

3.2. Appearance, color and smell of peptone are determined organoleptically.

3.3. Determination of the concentration of hydrogen ions (pH)

3.3.1. Equipment and reagents



pH meter;

laboratory scales;

glass flask according to GOST 1770-74;

GOST 1770-74;

paper filters;

distilled water according to GOST 6709-72.

3.3.2. Conducting a test

A weighed portion of peptone weighing 1 g, weighed with an error of not more than 0.02 g, is placed in a flask and dissolved in 99 cm 3 of boiled and cooled distilled water. A slight precipitate is allowed in the solution, which is filtered through a paper filter. In a transparent filtrate of a 1% peptone solution, the concentration of hydrogen ions (pH) is determined in accordance with the rules attached to the pH meter.

3.4. Determination of the mass fraction of insoluble impurities

(Changed edition, Rev. N 2).

3.4.1. Equipment, materials and reagents

For testing use:

analytical balances;

drying cabinet at 105 °C;

GOST 25336-82;

desiccator according to GOST 25336-82;

glass measuring cylinder with a capacity of 100 ml in accordance with GOST 1770-74;

cotton-gauze corks;

paper filters;

sodium oxide hydrate according to GOST 4328-77, 0.1 n. solution;

distilled water according to GOST 6709-72.

3.4.2. Conducting a test

A weighed portion of peptone weighing 5 g, weighed with an error of not more than 0.0002 g, is placed in a flask, 95 cm3 of distilled water are added and thoroughly mixed, achieving maximum dissolution of the peptone. The prepared 5% peptone solution is filtered through a paper filter, previously dried in a bottle to constant weight and weighed with an error of not more than 0.0002 g. After filtering the peptone solution, the filter is washed with 50 cm3 of distilled water, placed in a bottle and dried to constant weight. at 105 °C. The first weighing is carried out after drying the filter for 3 hours, subsequent weighings - after 30 minutes of drying. Drying is considered complete if the decrease in weight during two subsequent weighings does not exceed 0.0002 g.

The filtered solution of 5% peptone is diluted 5 times with distilled water. The resulting 1% peptone solution is adjusted to 0.1 N. sodium hydroxide solution to pH 7.7 and boil for 5 minutes. The solution should remain clear, without the formation of a precipitate.

After boiling, a 1% peptone solution is poured into test tubes of 7-8 cm, closed with cotton plugs and kept in an autoclave for 30 minutes at 0.15 MPa. The solution should remain clear, without the formation of a precipitate.

3.4.3. Results processing

The mass fraction of insoluble impurities () as a percentage is calculated by the formula

where is the mass of peptone taken to prepare 100 g of a 5% peptone solution, g;

- mass of weighing bottle with filter before filtration, g;

- weight of bottle and filter with sediment after drying, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.1%.

(Changed edition, Rev. N 2).

3.5. Determination of the mass fraction of moisture

(Changed edition, Rev. N 2).

3.5.1. Equipment

For testing use:

analytical balances;

laboratory drying cabinet at 105 °C;

cups for weighing (bottle bags) in accordance with GOST 25336-82;

desiccator according to GOST 25336-82.

3.5.2. Conducting a test

A weighed portion of peptone weighing 1.5-2.0 g, weighed with an error of not more than 0.0002 g, is placed in a weighing bottle, previously dried to a constant weight. The weighed bottle is placed in an oven and dried to constant weight at 105°C.

The first weighing is carried out after 1 hour, the next - after 30 minutes of drying. Drying is considered complete if the decrease in weight during two subsequent weighings does not exceed 0.0002 g.

3.5.3. Results processing

The mass fraction of moisture () as a percentage is calculated by the formula

where is the mass of the peptone sample before drying, g;

is the weight of the peptone sample after drying, g.



(Changed edition, Rev. N 2).

3.6. Determination of the mass fraction of sulfonated ash

(Changed edition, Rev. N 2).

3.6.1. Equipment and reagents

For testing use:

analytical balances;

heating element (tile, burner);

muffle furnace;

porcelain crucibles according to GOST 9147-80;

sulfuric acid according to GOST 4204-77, concentrated.

3.6.2. Conducting a test

A portion of peptone weighing 1.0-1.5 g, weighed with an error of not more than 0.0002 g, is placed in a crucible, previously calcined to constant weight. 1 cm3 of concentrated sulfuric acid is added to the crucible with a peptone weight and the mixture is carefully heated on a hot plate until excess sulfuric acid is removed, after which the crucible is transferred to a muffle furnace and ashing is continued at a temperature of 500-600 ° C to a constant mass of ash.

3.6.3. Results processing

The mass fraction of sulfonated ash () as a percentage is calculated by the formula

where is the mass of peptone before calcination, g;

- mass of ash after calcination, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

(Changed edition, Rev. N 2).

3.7. Determination of the mass fraction of true peptone

3.7.1. Equipment and reagents

For testing use:

photoelectric colorimeter or spectrophotometer;

desktop centrifuges type OPn-8-U4,2;

pipettes with a capacity of 1 and 5 cm according to GOST 29169-91;

glass test tubes according to GOST 25336-82;

sodium oxide hydrate according to GOST 4328-77, 10% solution;

copper sulfate according to GOST 4165-78, 2% solution;

distilled water according to GOST 6709-72.

(Changed edition, Rev. N 2).

3.7.2. Preparing for the test

Construction of a calibration graph

The abscissa shows the concentration of true peptone in the solution, the ordinate shows the value of the optical density () of this solution according to the data given in Table 2. The values ​​of optical density () given in Table 2 were obtained by measuring on a photoelectric colorimeter at a wavelength of 540 nm solutions of a specially prepared series of peptone containing a minimum of free protein and amino acids and a maximum of peptides.

table 2

3.7.3. Conducting a test

True peptone is determined in a 0.5% solution of the tested peptone. To do this, a 5% peptone solution prepared in accordance with clause 3.4.2 is diluted 10 times with water (0.5 ml of a 5% peptone solution and 4.5 cm3 of distilled water).

In a test tube containing 5 cm3 of a 0.5% solution of the test peptone, add 0.5 cm3 of a 10% solution of sodium hydroxide and 0.5 cm3 of a 2% solution of copper sulphate. At the same time, a control is placed, for which 5 cm3 of water, 0.5 cm3 of a 10% solution of sodium hydroxide and 0.5 cm3 of a 2% solution of copper sulphate are poured into a test tube.

The mixture in test tubes is well mixed and centrifuged for 10 minutes at a rotation frequency of 5000-6000 minutes. The color intensity is measured on a photoelectrocolorimeter at a wavelength of 540-560 nm or on a spectrophotometer at a wavelength of 540 nm in cuvettes with a working length of 10 mm. The concentration of true peptone in the sample is determined by the calibration curve.

(Changed edition, Rev. N 2

3.7.4. Results processing

The mass fraction of true peptone () as a percentage is calculated by the formula

where is the content of true peptone in the test solution, determined according to the calibration curve,%;

- mass of peptone (preparation) contained in 100 g of a 0.5% solution, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 5%.

(Changed edition, Rev. N 2).

3.8. Determination of the mass fraction of total nitrogen

(Changed edition, Rev. N 2).

The content of total nitrogen is determined by the Kjeldahl method or by the Kjeldahl method in combination with isothermal distillation in Conway cups.

3.8.1. Determination of the mass fraction of total nitrogen by the Kjeldahl method

(Changed edition, Rev. N 2).

3.8.1.1. Equipment and reagents

For testing use:

analytical balances;

GOST 25336-82;

apparatus for stripping ammonia;

pipettes with a capacity of 1, 2, 5 cm in accordance with GOST 29169-91;

burettes with a capacity of 25 or 50 cm3 with a division value of 0.05 cm3 in accordance with GOST 29252-91;

distilled water according to GOST 6709-72;

sulfuric acid according to GOST 4204-77, concentrated and 0.1 n. solution;

sodium oxide hydrate according to GOST 4328-77, 30-33% and 0.1 n. solutions prepared with pre-boiled distilled water;

hydrogen peroxide according to GOST 10929-76;

methyl red;

methyl blue;

GOST 5962-67 *;
________________
* On the territory of the Russian Federation, GOST R 51652-2000 applies. Here and further. - Database manufacturer's note.

methyl orange, 0.1% solution.

3.8.1.2. Conducting a test

1 ml of a 5% peptone solution is placed in a Kjeldahl flask and 2 ml of concentrated sulfuric acid are added. A glass pear-shaped canister or a small funnel with a sealed tip is inserted into the neck of the flask and the sample is first mineralized over low heat, then the degree of heating is increased, preventing the boiling mixture from being thrown out into the narrow part of the flask. Mineralization is carried out in the presence of a hydrogen peroxide catalyst. The catalyst is added after the start of boiling by 0.5 cm every 15-20 minutes until the solution is completely discolored.

After cooling, the contents of the flask are transferred from 10-12 ml of distilled water to a distillation flask. The completeness of the transfer is checked by the methyl orange indicator. The last portion of flushing water should remain yellow.

In the receiving flask of the Kjeldahl apparatus, pour 20 ml of 0.1 N. sulfuric acid solution and 2 drops of Tashiro indicator. The Tashiro indicator is prepared by dissolving 0.4 g of methyl red and 0.2 g of methyl blue in 200 ml of 96% ethanol.

The stripping flask is connected to a condenser and a vaporizer. The contents of the stripping flask are neutralized by methyl orange indicator with 30-33% sodium hydroxide solution. The distillation is carried out until about 70 cm3 of solution is collected in the receiving flask.

The contents of the receiving flask are titrated with 0.1 N. sodium hydroxide solution until the color of the solution changes from purple to green. At the same time, they put a control experiment on reagents, for which, instead of a peptone solution, 1 cm3 of distilled water is taken into the flask for mineralization

3.8.1.3. Results processing

where is the amount of 0.1 n. sulfuric acid solution poured into a receiving flask, cm;

- correction factor to the titer 0.1 n. sulfuric acid;

- the amount of 0.1 n. sodium hydroxide solution used for titration of the test or control sample, cm;


- mass of peptone contained in 1 cm 5% solution taken for analysis, g;

- the amount of nitrogen equivalent to 1 cm 0.1 N. sulfuric acid solution,

The final test result is the difference between the arithmetic mean of the results of two parallel determinations in experimental and control samples. Permissible discrepancies between the results of two parallel determinations should not exceed 0.2%.

(Revised edition, Rev. N

3.8.2. Determination of the mass fraction of total nitrogen by the Kjeldahl method in combination with isothermal distillation in Conway dishes

(Changed edition, Rev. N 2).

3.8.2.1 Apparatus and reagents

For testing use:

microburettes;

pipettes with a capacity of 1, 2, 5, 10 cm in accordance with GOST 29169-91;

Conway cups;

Kjeldal flasks with a capacity of 50-100 cm 3 according to GOST 25336-82;

volumetric flasks with a capacity of 100 cm3 in accordance with GOST 1770-74;

vaseline according to GOST 5774-76;

hydrogen peroxide according to GOST 10929-76;

methyl red;

methylene blue;

ethyl alcohol rectified according to GOST 5962-67;

sulfuric acid GOST 4204-77, concentrated and 0.025 n. solution;

sodium oxide hydrate (caustic soda) according to GOST 4328-77, 40% and 0.025 n. solutions;

distilled water according to GOST 6709-72.

3.8.2.2. Preparing for the test

Preparation of the Tashiro indicator according to clause 3.8.1.2.

3.8.2.3. Conducting a test

5 ml of a 5% peptone solution are mineralized with heating in a Kjeldahl flask with 10 ml of sulfuric acid (concentrated) until a colorless solution is obtained. 0.5 cm of perhydrol is used as a catalyst. After mineralization, the sample was quantitatively transferred into a 100 ml volumetric flask and made up to the mark with distilled water. Pour exactly 2 cm of 0.025 N into the inside of the Conway dish. sulfuric acid solution. To prevent the liquid used from spreading, the cup is set obliquely. Then close the lid, smeared with petroleum jelly, leaving a small gap, through which 1 cm of the test solution after mineralization is introduced into the outer part of the cup with a pipette. The cup is moved so that a gap of 5-6 mm wide remains on the opposite side, and 4 cm of a 40% alkali solution is quickly poured through it. Close the cup tightly with a lid, carefully mix the contents in a circular motion and leave for 15-18 hours. sodium hydroxide solution according to the Tashiro indicator. In parallel, a control sample is placed, for which distilled water is placed in the outer part of the Conway cup instead of the test solution.

3.8.2.4. Results processing

The mass fraction of total nitrogen () as a percentage is calculated by the formula

where is the amount of 0.025 n. sodium hydroxide solution used for titration of the control sample, cm;

- the amount of 0.025 n. sodium hydroxide solution used for titration of the test sample, cm;

- correction factor to the titer 0.025 n. sodium hydroxide solution;

- the amount of nitrogen equivalent to 1 cm 0.025 N. sodium hydroxide solution, g;

- mass of peptone taken to obtain a 5% solution, g;

- the amount of the drug taken for mineralization, ml;

- the amount of the test solution, see

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

(Revised edition, Rev.

3.9. Determination of the mass fraction of nitrogen of amino groups of amino acids and lower peptides

(Changed edition, Rev. N 2).

3.9.1. Equipment and reagents

For testing use:

pH meter;

laboratory glasses according to GOST 25336-82;

pipettes with a capacity of 1, 2 and 20 cm in accordance with GOST 29169-91;

burettes with a capacity of 5 and 10 cm3 with a division value of 0.05 cm3 in accordance with GOST 29252-91;

formalin technical according to GOST 1625-89;

sodium oxide hydrate according to GOST 4328-77, 0.1 n. solution;

sulfuric acid according to GOST 4204-77, 0.1 g * solution;

phenolphthalein according to NTD, 1% alcohol solution;

distilled water according to GOST 6709-72.
________________

3.9.2. Preparing for the test

Immediately before work, the formol mixture is prepared. To do this, add 1 cm of a 1% solution of phenolphthalein to 50 cm of formalin and 0.1 g * of sodium hydroxide solution to bring the color of the mixture to slightly pink.
________________
* Corresponds to the original. - Database manufacturer's note.

3.9.3. Conducting a test

To 2 cm of a transparent 5% peptone solution prepared in accordance with clause 3.4.2, add 18 cm of distilled water and adjust the pH of the mixture to 7.0 potentiometrically 0.1 N. a solution of sodium hydroxide or sulfuric acid. Then 2 ml of the formol mixture is poured and, with constant stirring, titrated potentiometrically with 0.1 N. sodium hydroxide solution to pH 9.2.

At the same time, a control is placed on the reagents, for which, instead of 2 cm3 of 5% peptone, 2 cm3 of distilled water are taken.

3.9.4. Results processing

The mass fraction of nitrogen of amino groups of amino acids and lower peptides in peptone () as a percentage is calculated by the formula

where is the amount of 0.1 n. sodium hydroxide solution used for titration of the test sample, cm;

- the amount of 0.1 n. sodium hydroxide solution used for titration of the control sample, cm;

- correction factor to the titer 0.1 n. sodium hydroxide solution;

- the amount of nitrogen equivalent to 1 cm 0.1 N. sodium hydroxide solution, g;

- mass of peptone contained in 2 cm3 of 5% peptone, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

(Revised edition, Rev. N

3.10. Determination of free protein

(Changed edition, Rev. N 2).

3.10.1. Equipment and reagents

For testing use:

photoelectric colorimeter;

pipettes with a capacity of 5 and 10 cm in accordance with GOST 29169-91;

glass test tubes according to GOST 25336-82;

trichloroacetic acid 20% solution.

3.10.2. Conducting a test

To 5 cm3 of the 5% peptone solution filtrate prepared in accordance with clause 3.4.2, add 5 cm3 of a 20% trichloroacetic acid solution and mix well. After 5 minutes, the optical density of the mixture is determined on a photoelectric colorimeter at a wavelength of 630 nm (red light filter) in cuvettes with a working length of 10 mm against the same peptone solution diluted 1:1 with distilled water.

3.10.3. Results processing

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.05%.

3.11. Determination of the mass fraction of chlorides

(Changed edition, Rev. N 2).

3.11.1. Equipment and reagents

For testing use:



heating element (burner, stove);

pipettes with a capacity of 1, 2, 10, 20 and 25 cm in accordance with GOST 29169-91;

burettes with a capacity of 25 cm3 with a division value of 0.05 cm3 in accordance with GOST 29252-91;

laboratory glasses according to GOST 25336-82;

distilled water according to GOST 6709-72;

silver nitrate according to GOST 1277-75, 0.1 n. solution;

ferroammonium alum according to NTD, saturated solution;

potassium thiocyanate according to NTD or ammonium thiocyanate 0.1 n. solution;

nitric acid according to GOST 4461-77, concentrated.

(Changed edition. Rev. N 2).

3.11.2. Conducting a test

To 20 ml of a 1% peptone solution prepared in accordance with clause 3.4.2, add 30 ml of distilled water, 1 ml of concentrated nitric acid and 10 ml of 0.1 N. silver nitrate solution. The solution is heated to boiling and quickly cooled by immersing the beaker of the solution in a vessel of water at room temperature. After cooling the mixture, add 2 ml of a saturated solution of iron ammonium alum (indicator) and titrate with 0.1 N. a solution of potassium thiocyanate (or ammonium thiocyanate) until a brick-red color appears.

3.11.3. Results processing

The mass fraction of chlorides () as a percentage is calculated by the formula

where is the amount of 0.1 n. a solution of silver nitrate, taken into a beaker with a sample, cm;

- correction factor to the titer 0.1 n. silver nitrate solution;

- the amount of 0.1 n. solution of potassium thiocyanate or ammonium thiocyanate used for titration of the sample, cm;

- correction factor to the titer 0.1 n. a solution of potassium thiocyanate or ammonium thiocyanate;

- the amount of chlorides (chlorine ion), equivalent to 1 ml of 0.1 N. silver nitrate solution, g;

- the mass of peptone contained in 20 cm3 of a 1% solution, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.1%.

(Revised edition, Rev. N

3.12. Determination of salts of heavy metals

3.12.1. Equipment and reagents

For testing use:

technical scales with a weighing error of not more than 0.01 g;

heating element (stove, burner); water distilled with concentrated sulfuric acid and carefully ashed first on a stove or burner, and then in a muffle furnace at 500-600 ° C until a homogeneous light gray ash is obtained.

After cooling the crucible, the ash is poured into 1 cm3 of a saturated solution of ammonium acetate and diluted with 5 cm3 of distilled water. The contents of the crucible are filtered into the cylinder through a paper filter. The crucible and filter are washed with distilled water, bringing the volume of the solution in the cylinder to 10 cm3.

The reaction of the filtrate in the cylinder is brought to neutral or slightly acidic by litmus by adding dropwise 10% acetic acid or 10% sodium hydroxide solution. Then add 1 ml of 10% acetic acid, 5 drops of a saturated solution of sodium sulfide and mix. After 5 minutes, the solution should not have a precipitate and a yellow color of the solution.

3.13. Determination of the presence of free indole

3.13.1. Equipment and reagents

For the test, apply: 0.1% solution;

sulfuric acid according to GOST 4202-75 *, concentrated.
_________________
* Reference to the standard corresponds to the original. - Database manufacturer's note.

3.13.2. Conducting a test

Add 2 ml of amyl alcohol to a test tube with 10 cm3 of 1% peptone solution, close the tube with a cork, shake well, and add 1 cm3 of 0.1% potassium nitrite solution and 1 cm3 of concentrated sulfuric acid. The mixture is gently stirred and allowed to settle.

No pink coloration should appear in the alcohol layer. The appearance of a pink coloration of the alcohol layer indicates the presence of free indole in the peptone.

3.14. Determination of the ability to support microbial growth

3.14.1. Equipment, materials and reagents

For testing use:

technical scales with a weighing error of not more than 0.01 g;

pH meter;

thermostat at 37 °C;

autoclave;

photoelectric colorimeter;

test tubes according to GOST 25336-82;

cotton-gauze corks;

micropipettes with a capacity of 0.2 cm;
nutrient media (meat water according to
To prepare MPA, 1% of the test peptone, 0.5% sodium chloride, 2% microbiological agar are added to the meat water and the pH of the medium is set to 7.4.

MPB and MPA are filtered, poured into 8 cm test tubes, closed with cotton-gauze stoppers and parchment caps and sterilized in an autoclave for 30 minutes at 0.15 MPA.

3.14.3. Conducting a test

Test tubes with sterile BCH and oblique MPA are inoculated with daily control cultures. Each culture is inoculated with a 0.2 cm micropipette into three test tubes with MPB and MPA. Incubation is carried out for 24 hours at 37-38 °C.

The typical growth of daily cultures is determined on the MPA visually and under a microscope. Growth should be typical of each strain* of microorganisms. The growth intensity is evaluated in the BCH on a photoelectrocolorimeter at a wavelength of 620-640 nm in a cuvette with a working length of 5 mm.
________________
* The text corresponds to the original. - Database manufacturer's note.

4. PACKAGING, PACKAGING, MARKING, TRANSPORTATION AND STORAGE

4.1. Peptone is packaged with a net weight of 250 to 1000 g in cans according to GOST 5981-88 or, by agreement with the consumer, with a net weight of 30 kg in type II plywood barrels according to NTD with a polyethylene film lining in accordance with GOST 10354-82. The cans are hermetically sealed.

(Changed edition, Rev. N 1).

4.2. The jar is labeled with:

name of the manufacturer;

name of the drug;

series numbers;

net weight;

storage conditions;

expiration date;
the name of the manufacturer and its trademark;

name of the drug;

series numbers;

manufacturing dates;

gross and net weights;

storage conditions;

expiration date;

designations of this standard.

A handling sign "Afraid of dampness" is applied to each box or barrel.

(Changed edition, Rev. N 1).

4.5. Peptone is stored in a dry room at a temperature not exceeding 30 °C. The shelf life of peptone in hermetically sealed jars is 3 years, in plywood barrels - 1 year from the date of manufacture.

4.6. Peptone is transported by all means of transport in compliance with the rules for the carriage of goods in force on this type of transport.

Strengthening of packages in overpacks when transporting goods by rail in accordance with GOST 24597-81 and GOST 26663-85.

(Changed edition, Rev. N 1).

Electronic text of the document
prepared by Kodeks JSC and verified against:
official publication
M.: IPK Standards Publishing House, 1995

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Price 5 kop.

Official edition

USSR STATE COMMITTEE ON STANDARDS Moscow

UDC 547.984.3: 006.354 Group H18

STATE STANDARD OF THE UNION OF THE SSR

PEPTONE DRY ENZYMATIVE FOR BACTERIOLOGICAL PURPOSES

Specifications

Dry fermentation pepton for bacteriological objects. technical conditions

By the Decree of the State Committee of Standards of the Council of Ministers of the USSR dated April 29, 1976 No. 1002, the validity period is established

from 01.01-1977 to 01.01. 1982

Non-compliance with the standard is punishable by law

This standard applies to enzymatic peptone for bacteriological purposes, obtained from the rumen and lettuce of cattle, sheep and goats, as well as from the stomachs of pigs using the mucous membrane of the stomach and pancreas.

1. TECHNICAL REQUIREMENTS

1.1. Enzymatic peptone for bacteriological purposes must be manufactured in accordance with the requirements of this standard according to the technological rules approved in the prescribed manner.

1.2. Enzymatic peptone for bacteriological purposes in terms of physicochemical and biological indicators must comply with the requirements and standards specified in Table. one.

Table 1

© Standards Publishing, 1981

Page 10 GOST 1380J-76

titrated with 0.025 N. sodium hydroxide solution according to the Tashiro indicator. In parallel, a control sample is placed, for which distilled water is placed in the outer part of the Conway cup instead of the test solution.

3.8.2.4. Results processing

V 0.00035(T-Vi)K- 100-100- ICO * 5== -5^1- ’

where V is the number of 0.025 n. sodium hydroxide solution used for titration of the control sample, ml;

V t - the amount of 0.025 n. sodium hydroxide solution used for titration of the test sample, ml;

K - correction factor to the titer 0.025 n. sodium hydroxide solution;

0.00035 is the amount of nitrogen equivalent to 1 ml of 0.025 N. sodium hydroxide solution, g;

5-mass of peptone taken to obtain a 5% solution, g;

5 - the amount of the drug taken for mineralization, ml; 1 - the amount of the test solution, ml.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

3.9. Determination of nitrogen content of amino groups of amino acids and lower peptides

3.9.1. Equipment and reagents

pH meter;

3.9.2. Preparing for the test

Immediately before work, the formol mixture is prepared. To do this, add 1 ml of a 1% solution of phenolphthalein and 0.1 N of formalin to 50 ml of formalin. with a solution of sodium hydroxide, bring the color of the mixture to slightly pink.

3.9.3. Conducting a test

To 2 ml of a clear 5% peptone solution prepared in accordance with paragraph 3.4.2, add 18 ml of distilled water and adjust the pH of the mixture to 7.0 potentiometrically 0.1 N. a solution of sodium hydroxide or sulfuric acid. Then, 2 ml of the formol mixture is added and, with constant stirring, titrated potentiometrically with 0.1 N. sodium hydroxide solution to pH 9.2.

At the same time put the control on the reagents, for which instead of 2 ml of 5% peptone take 2 ml of distilled water.

3.9.4. Results processing

(V- -0.010114. ju

where V is the amount of 0.1 n. sodium hydroxide solution used for titration of the test sample, ml;

V) - the amount of 0.1 n. sodium hydroxide solution used for titration of the control sample, ml;

K - correction factor to the titer of 0.1 n. sodium hydroxide solution;

0.0014 - the amount of nitrogen equivalent to 1 ml of 0.1 n. sodium hydroxide solution, g;

0.1 mass of peptone contained in 2 ml of 5% peptone, g

3.10. Determination of the content of free protein

3.10.1. Equipment and reagents

For testing use:

photoelectric colorimeter;

pipettes with a capacity of 5 and 10 ml;

trichloroacetic acid 20% solution.

3.10.2. Conducting a test

To 5 ml of the filtrate of a 5% peptone solution prepared in accordance with paragraph 3.4.2, add 5 ml of a 20% solution of trichloroacetic acid and mix well. After 5 minutes, the optical density of the mixture is determined on a photoelectric colorimeter at a wavelength of 630 nm (red light filter) in cuvettes with a working length of 10 mm against the same peptone solution diluted 1: 1 with distilled water.

3.10.3. Results processing

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.05%.

3.11. Determination of chloride content

3.11.1. Equipment and reagents

For testing use:

technical scales with a weighing error of not more than 0.01 g; heating element (burner, stove); pipettes with a capacity of 1, 2, 10, 20 and 25 ml; burettes with a capacity of 25 ml with a division value of 0.05 ml; laboratory glasses according to GOST 10394-72; distilled water according to GOST 6709-72; silver nitrate according to GOST 1277-75, 0.1 n. solution; ferroammonium alum according to GOST 4205-77, saturated solution;

3.12. Determination of salts of heavy metals

3.12.1. Equipment and reagents

For testing use:

technical scales with a weighing error of not more than 0.01 g; heating element (tile, burner); muffle furnace;

pipettes with a capacity of 1, 2 and 10 ml;

rubber cone plugs according to GOST 7852-76;

amyl alcohol;

3.14.2. Preparing for the test

Meat-peptone broth (MPB) is prepared in accordance with the requirements of GOST 20730-75 and meat-peptone agar (MPA). To prepare the MPB, 1% of the tested peptone, 0.5% sodium chloride are added to the meat water and the pH of the medium is set to 7.4.

To prepare MPA, 1% of the test peptone, 0.5% sodium chloride, 2% microbiological agar are added to meat water and the pH of the medium is set to 7.4.

MPB and MPA are filtered, poured into test tubes in 8 ml, closed with cotton gauze stoppers and parchment caps and sterilized in an autoclave for 30 minutes at 0.15 MPa.

3.14.3. Conducting a test

Test tubes with sterile BCH and oblique MPA are inoculated with daily control cultures. Inoculation of each culture is carried out with a micropipette of 0.2 ml in three test tubes with MPB and MPA.

GOST 13S05-76 Page fifteen

Incubation is carried out for 24 hours at 37-38°C.

The typicality of the growth of daily cultures is determined on the MPA visually and under a microscope. Growth should be typical for each microorganism stamp. The growth intensity is evaluated in the BCH on a photoelectrocolorimeter at a wavelength of 620-640 nm in a cuvette with a working length of 5 mm.

4. PACKAGING, PACKAGING, MARKING, TRANSPORTATION AND STORAGE

4.1. Peptone is packaged with a net weight of 250 to 1000 g in cans in accordance with GOST 5981-71 or, by agreement with the consumer, with a net weight of 30 kg in plywood barrels in accordance with GOST 5958-79 with a polyethylene film lining in accordance with GOST 10354-73. The cans are hermetically sealed.

4.2. A label is attached to the jar indicating: the name of the manufacturer; name of the drug;

series numbers; net weight; storage conditions; expiration date;

4.3. Banks with peptone are packed in plank non-separable boxes for canned food* in accordance with GOST 13358-72 or in corrugated cardboard boxes for canned food and food liquids in accordance with GOST 13516-72.

4.4. Each box and barrel is marked in accordance with GOST 14192-77 with additional data:

name of the drug;

series numbers;

manufacturing dates;

gross and net weights;

storage conditions;

expiration date;

designations of this standard.

A warning sign "Afraid of dampness" is applied to each box or barrel.

4.5. Peptone is stored in a dry room at a temperature not exceeding 30°C. The shelf life of peptone in hermetically sealed jars is 3 years, in plywood barrels - 1 year from the date of manufacture.

4.6. Peptone is transported by all modes of transport.

4.7. In case of violation of the sealing of the container, peptone is stored in a tightly closed vessel or plastic bag in a dry room at a temperature of 0 to 8 ° C for no more than 1 month.

Clauses 3.6.1, 3.8.1.1, 3.8.2.1, 3.9.1, 3.12.1. Replace reference: GOST 4204-66 with GOST 4204_77.

Paragraphs 3.7.1, 3.8.1.1, 3.8.2.1, 3.9.1, 3.10.3, 3.11.1, 3.13.1, 3.14.1 regarding pipettes and burettes shall be supplemented with the reference: “according to GOST 20292-74”.

Clause 3.7.1, Replace the words and reference: “glass funnels” with “glass funnels according to GOST 8613-75”; GOST 4165-68 to GOST 4165-78.

(Continued see page 170J

Paragraph 4.1 after the words "plywood barrels" shall be supplemented with the words: "type II"; replace the link: GOST 5958-70 with GOST 5958-79.

Clause 4.4. Replace the words: “Each box and barrel is marked in accordance with GOST 14192-71 with additional data” for “Transport marking in accordance with GOST 14192-77 with additional data”; “name of the manufacturer” to “name of the manufacturer and its trademark”; "precautionary" to "manipulative".

Paragraph 4.6 shall be supplemented with the words: “in compliance with the rules for the carriage of goods in force on this type of transport.

Strengthening of packages in overpacks when transporting goods by rail in accordance with GOST 21929-76 and GOST 15901-70.

(IUS No. 3 1982)

Paragraph 3.7.1 shall be stated in a new wording:

(Continued, see p. 290) (Continued change to GOST 13805-76>

"3.7.1, Apparatus and reagents

For testing use:

photoelectric colorimeter or spectrophotometer;

desktop centrifuges type OPn-6-U4,2,

pipettes with a capacity of 1 and 5 cm 3 according to GOST 20292-74;

From the total sample of enzymatic peptone, four samples are isolated for analysis, each weighing at least 100 g. The selected samples are placed in clean, dry jars with ground stoppers. Corks are filled with paraffin or resin.

3.1.2. Enzymatic peptone sample jars are labeled with:

name of the manufacturer;

name of the drug;

manufacturing dates;

series numbers;

date of sampling;

designation of this standard.

Two samples are used for analysis, and two are left in the archives of the State Comptroller for 4 years.

3.2. Appearance, color and smell of peptone are determined organoleptically.

3.3. Determination of the concentration of hydrogen ions (pH)

3.3.1. Equipment and reagents

laboratory scales;

GOST 1770-74;

paper filters;

3.3.2. Conducting a test

A weighed portion of peptone weighing 1 g, weighed with an error of not more than 0.02 g, is placed in a flask and dissolved in 99 ml of boiled and cooled distilled water. A slight precipitate is allowed in the solution, which is filtered through a paper filter. In a transparent filtrate of a 1% peptone solution, the concentration of hydrogen ions (PH) is determined in accordance with the rules attached to the pH meter.

3.4. Determination of the content of insoluble impurities

3.4.1. Equipment, materials and reagents Drying cabinet at 105°С;

GOST 7148-70; desiccator according to GOST 6371-73;

glass measuring cylinder with a capacity of 100 ml in accordance with GOST 1770-74;

paper filters;

3.4.2. Conducting a test

A weighed portion of peptone weighing 5 g, weighed with an error of not more than 0.0002 g, is placed in a flask, 95 ml of distilled water are added and thoroughly mixed, achieving maximum dissolution of peptone. The prepared 5% peptone solution is filtered through a paper filter, previously dried in a bottle to constant weight and weighed with an error of not more than 0.0002 g. After filtering the peptone solution, the filter is washed with 50 ml of distilled water, placed in a bottle and dried to constant weight. at 105°C. The first weighing is carried out after drying the filter for 3 hours, subsequent weighings - after 30 minutes of drying. Drying is considered complete if the decrease in weight during two subsequent weighings does not exceed 0.0002 g.

The filtered solution of 5% peptone is diluted 5 times with distilled water. The resulting 1% peptone solution is adjusted to 0.1 N. sodium hydroxide solution to pH 7.7 and boil for 5 minutes. The solution should remain clear, without the formation of a precipitate.

After boiling, a 1% peptone solution is poured into test tubes of 7-8 ml, closed with cotton plugs and kept in an autoclave for 30 minutes at 0.15 MPa. The solution should remain clear, without the formation of a precipitate.

3.4.3. Results processing

_ (/722-P1\) * lllClfO

where m is the mass of peptone taken to prepare 100 g of a 5% peptone solution, g; mi is the weight of the weighing bottle with the filter before filtration, g; t 2 - weight of bottle and filter with sediment after drying, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.1%.

3.5. Determination of moisture content 3.5.1. Equipment

To carry out the test, the following are used: analytical balances;

laboratory drying cabinet at 105°С;

cups for weighing (bottle bags) in accordance with GOST 7148-70;

3.5.2. Conducting a test

A weighed portion of peptone weighing 1.5-2.0 g, weighed with an error of not more than 0.0002 g, is placed in a weighing bottle, previously dried to a constant weight. The weighed bottle is placed in an oven and dried to constant weight at 105°C.

The first weighing is carried out after 1 hour, the next - after 30 minutes of drying. Drying is considered complete if the decrease in weight during two subsequent weighings does not exceed 0.0002 g.

3.5.3. Results processing

y (m-! P \) * 1 | 0U

where m is the weight of the peptone sample before drying, g; rn is the weight of the peptone sample after drying, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

3.6. Determination of the content of sulfonated ash

3.6.1. Equipment and reagents

For testing use:

analytical balances;

heating element (tile, burner);

muffle furnace;

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 0.2%.

3.7. Determination of true peptone content

3.7.1. Equipment and reagents

To carry out the test, apply: photoelectrocolorimeter or spectrophotometer; pipettes with a capacity of 1 and 5 ml; glass test tubes according to GOST 10515-75; glass funnels; paper filters:

3.7.2. Preparing for the test Building a calibration curve

The concentration of true peptone in the solution is plotted along the abscissa, and the optical density (E) of this solution is plotted along the ordinate, according to the data given in Table. 2. Given in table. 2 values ​​of optical density (E) were obtained by measuring on a photoelectrocolorimeter at a wavelength of 540 nm solutions of a specially prepared peptone series containing a minimum of free protein and amino acids and a maximum of peptides.

table 2

Concentration of true peptone in solution, %

Optical density (£) solution

0.560 0R80 0.190

3.7.3. Conducting a test

True peptone is determined in a 0.5% solution of the tested peptone. To do this, a 5% peptone solution prepared in accordance with paragraph 3.4.2 is diluted 10 times with water (0.5 ml of a 5% peptone solution and 4.5 ml of distilled water).

In a test tube containing 5 ml of a 0.5% solution of the test peptone, add 0.5 ml of a 10% solution of sodium hydroxide and 0.5 ml of a 2% solution of copper sulfate. At the same time, a control is placed, for which 5 ml of water is poured into the test tube,

0.5 ml of 10% sodium hydroxide solution and 0.5 ml of 2% copper sulfate solution.

The mixture in test tubes is well mixed and after 2 min is filtered through paper filters. The color intensity is measured on a photoelectric colorimeter at a wavelength of 540-560 nm or on a spectrophotometer at a wavelength of 540 nm in cuvettes with a working length of 10 mm. The concentration of true peptone in the sample is determined by the calibration curve.

3.7.4. Results processing

y _ A-11Y00 Az 0.5

0.5 mass of peptone (preparation) contained in 100 g of a 0.5% solution, g.

The arithmetic mean of the results of two parallel determinations is taken as the final test result. Permissible discrepancies between the results of parallel determinations should not exceed 5%.

3.8. Determination of total nitrogen content The total nitrogen content is determined by the Kjeldahl method or by the Kjeldahl method in combination with isothermal distillation in Conway dishes.

3.8.1. Determination of total nitrogen by the Kjeldahl method

3.8.1.1. Equipment and reagents

To carry out the test, the following are used: analytical balances;

Kjeldal flasks with a capacity of 50-100 ml according to GOST

apparatus for stripping ammonia; pipettes with a capacity of 1, 2, 5 ml;

burettes with a capacity of 25 or 50 ml with a division value of 0.05 ml; distilled water according to GOST 6709-72; sulfuric acid according to GOST 4204-77, concentrated and 0.1 n. solution;

3.8.2.2. Preparation for the test Preparation of the Tashiro indicator according to clause 3.8.1.2.

3.8.2.3. Conducting a test

5 ml of a 5% peptone solution are mineralized with heating in a Kjeldahl flask with 10 ml of sulfuric acid (concentrated) until a colorless solution is obtained. 0.5 ml of perhydrol is used as a catalyst. After mineralization, the sample was quantitatively transferred into a 100 ml volumetric flask and made up to the mark with distilled water. Pour exactly 2 ml of 0.025 N into the inside of the Conway dish. sulfuric acid solution. To prevent the liquid used from spreading, the cup is set obliquely. Then close the lid, smeared with petroleum jelly, leaving a small gap, through which 1 ml of the test solution after mineralization is pipetted into the outer part of the dish. The cup is moved so that there is a gap 5-6 mm wide on the opposite side, and 4 ml of a 40% alkali solution is quickly poured through it. Close the cup tightly with a lid, gently mix the contents in a circular motion and leave for 15-18 hours. Excess acid not bound to ammonia