Fundamentals of General and Pharmaceutical Hygiene - Dykyi I.L. 2003

Hygiene of Water and Water Supply
Sanitary requirements for the production, transportation, and storage of water used in pharmaceutical practice

In pharmaceutical practice, Water of varying degrees of purification is used (desalted (demineralized), purified (distilled), and water for injection). It is obtained from drinking tap water or natural water using specialized Methods such as distillation, reverse osmosis, Ion Exchange, or others. Improving the quality of water sourced from water supplies for production needs is referred to as water Treatment.

Demineralized water—freed from unwanted cations and anions—is used for washing glassware, ampoules, and auxiliary Materials, as well as for feeding water stills to produce purified water and water for injection.

Purified water (Aqua purificata) is used in the manufacture of injectable dosage forms and for the preparation of sterile solutions that are not required to be pyrogen-free. It must be chemically purified to the highest degree and comply with the requirements of current regulatory documentation.

Water for injection (Aqua ingectionibus), used in the preparation of parenteral solutions, must meet all the requirements established for purified water while additionally being sterile and pyrogen-free.

Pyrogens (from Greek pyr — fire and genes — producing) are substances, predominantly of bacterial origin (living microorganisms or their metabolic products), that, when administered parenterally into the body, are capable of causing a rapid increase in body Temperature, which may be accompanied by chills, headache, and in severe cases lead to cardiovascular dysfunction and collapse. Lipopolysaccharides of Gram-negative Bacteria pose a particular hazard; aside from their small size, which allows them to pass through the densest filters with pore sizes ranging from 0.005 to 0.001 µm, they are characterized by extreme thermostability.

To ensure the pyrogen-free status of water during its production, transportation, and storage, a full range of Sanitary and hygienic requirements aimed at preventing contamination by particulate matter and microorganisms must be observed. For the Production of purified water in pharmacy settings, a specially equipped room is designated, in which any type of work unrelated to water production is strictly prohibited. At the same time, absolute compliance with the technological regime and aseptic rules must be ensured.

Purified water may be stored for no more than 3 days.

Water for injection is stored for no more than 24 hours from the time of production at a temperature of 5–10 °C or 80–95 °C, at which point the GROWTH AND REPRODUCTION of microorganisms cease.

Storage tanks for water must be made of materials that do not affect its properties, protect it from mechanical impurities and microorganisms, exhibit resistance to thermal sterilization, and allow for efficient cleaning and disinfection.

Objective: to know the pharmacopeial requirements for purified water and water for injection, Methods of Analysis, and sanitary requirements for the production, transportation, and storage of purified water and water for injection; and to be able to develop recommendations for preventing pyrogenicity.

QUESTIONS FOR SELF-STUDY

1. Water as a solvent for medicinal substances.

2. Requirements for purified water and water for injection according to current regulatory documentation.

3. Pyrogens, their nature and properties.

4. Characteristics of pyrogens of bacterial origin (endotoxins).

5. Causes of pyrogenicity in parenteral solutions.

6. Sanitary requirements for The production of purified water and water for injection.

7. Sanitary requirements for the storage of purified water and water for injection.

8. Pharmacopeial methods of water analysis.

Task 1. Analysis of purified water for the absence of chlorides

Using a pipette, 10 ml of the test sample of purified water is placed into a test tube, 0.5 ml of a 2% silver nitrate solution is added, mixed gently, and after 5 minutes compared with a standard consisting of 10 ml of the standard solution and the same amount of reagent added to the water. After 15 minutes, the results of the reaction are evaluated.

The absence of opalescence indicates the absence of chlorides in the water.

Task 2. Analysis of purified water for the absence of nitrates and nitrites

Using a pipette, 5 ml of the test sample of purified water is placed into a test tube, 1 ml of freshly prepared 0.6% diphenylamine solution is carefully added, and the coloration of the solution is then evaluated.

The appearance of a blue coloration indicates the presence of nitrates and nitrites.

Task 3. Analysis of water for injection for sterility

The direct inoculation method of the water sample into liquid nutrient media is used. Test tubes are prepared in advance: one with 10 mL of thioglycollate medium (for the detection of bacteria) and the second with 10 mL of Sabouraud medium (for the detection of Fungi). Following aseptic techniques, the ampoule with water for injection is opened, and 1 mL is transferred using a sterile pipette into each test tube containing the aforementioned nutrient media. The water inoculation in the thioglycollate medium is incubated at a temperature of 35 °C, and in the Sabouraud medium at 25 °C for 14 days.

Microbial growth is indicated by visual signs such as turbidity, pellicle, sediment, or A change in the color of the medium. In the absence of the aforementioned changes, the tested sample of water for injection is considered to meet the sterility requirements.

Task 4. Analysis of water for injection for the absence of bacterial endotoxins (LAL test)

The method is based on gel formation resulting from the interaction of bacterial pyrogens (endotoxins) with the amoebocyte lysate from the Blood of horseshoe crabs (*Limulus polyphemus*) or other species (LAL test).

0.5 mL of the crab amoebocyte lysate and 0.5 mL of the test water sample are added to a test tube, followed by incubation in a thermostat at a temperature of 36 °C for 1 hour.

If pyrogens (bacterial endotoxins) are present in the test water, a gel is formed, which can be detected by inverting the test tube along its vertical axis by 180 °C.

PROTOCOL SCHEME

Topic of the practical Class.

1. Analysis of purified water for the absence of chlorides.

Volume of the test water sample ... .

Volume of silver nitrate ... .

Reaction time ... .

Result. Opalescence (was not) detected.

2. Analysis of purified water for the absence of nitrates and nitrites.

Volume of the test water sample ... .

Volume of diphenylamine solution ... .

Reaction time ... .

Result. Blue coloration (was not) detected.

3. Analysis of water for injection for sterility.

Volume of the test water sample ... .

Volume of thioglycollate medium ... .

Volume of Sabouraud medium ... .

Culture conditions (temperature, time) ... .

Result. No signs of bacterial growth (were) detected in the thioglycollate medium, and no signs of fungal growth (were) detected in the Sabouraud medium.

4. Testing water for injection for bacterial endotoxins (LAL test):

Principle of the method ... .

Amebocyte lysate from horseshoe crab blood (species) ... .

Volume of the test water sample ... .

Volume of amebocyte lysate ... .

Incubation temperature and time ... .

Result: when inverting the test tube by 180°, gel formation was (not) detected.

Conclusion. The results of chemical analysis of purified water indicate the absence (presence) of chlorides, nitrates, and nitrites. Water for injection (does not) meet pharmacopoeial requirements for sterility and apyrogenicity.

TASKS FOR INDEPENDENT WORK

1. Knowledge self-assessment test.

2. Solving situational problems.

TESTS

1. Waterborne infections can be transmitted by:

A. AIDS.

Б. Hepatitis A.

B. Tularemia.

Г. Hepatitis B.

2. Surface water bodies differ from interstratal waters by:

A. Higher oxygen content.

Б. Higher bacterial contamination.

B. More stable chemical composition.

Г. Higher mineralization.

3. Indicate the indirect indicator of biogenic pollution of a water body:

A. Saprobity.

Б. Nitrates.

B. Oxidizability.

Г. Fluorine.

4. Select the secondary indicator of fresh water pollution by domestic wastewater:

A. Chlorides.

Б. Sulfates.

B. Fluorine.

Г. Nitrates.

Д. Nitrites.

Е. Ammonia.

5. Select the secondary indicator of long-standing water pollution by domestic wastewater.

A. Fluorine.

Б. Sulfates.

B. Ammonia.

Г. Nitrates.

Д. Chlorides.

6. The tap water in settlement N. was found to contain a significantly higher than normal amount of fluorine, which is a risk factor for The Development of:

A. Dental caries.

Б. Fluorosis.

B. Kashin-Beck disease.

Г. Gout.

Д. Methemoglobinemia.

7. Drinking water from a decentralized source in settlement V. was found to contain a significantly higher than normal amount of strontium. What disease may occur in the population consuming this water?

A. Gout.

Б. Kashin-Beck disease.

B. Fluorosis.

D. Dental caries.

8. Tap water in settlement A. was found to have a chloride concentration significantly exceeding the standard, which is a risk factor for the development of:

A. Arterial Hypertension.

B. Kashin-Beck disease.

C. Urolithiasis.

D. Diabetes.

E. Dysentery.

9. Well water in settlement B. was found to have an increased nitrate concentration compared to the standard, which is a risk factor for the development of:

A. Kashin-Beck disease.

B. Methemoglobinemia.

C. Urolithiasis.

D. Arterial hypertension.

E. Infectious disease.

10. Well water in settlement N. exhibits hardness levels significantly exceeding the standard. What disease may occur in the population with prolonged consumption of such water?

A. Endemic goiter.

B. Fluorosis.

C. Urolithiasis.

D. Dental caries.

11. Methods of water disinfection:

A. Coagulation.

B. Filtration.

C. Chlorination.

D. Ozonation.

E. Sedimentation.

12. The main indicator of the reliability of water disinfection during chlorination:

A. Ammonia.

B. Residual chlorine.

C. Nitrates, nitrites.

D. Oxidizability.

E. Chlorides.

13. During the sanitary and Hygienic assessment of water bodies, the pollution index known as saprobity is used. Indicate the zone of minimum pollution:

A. Alpha-mesosaprobic.

B. Polysaprobic.

C. Beta-mesosaprobic.

D. Oligosaprobic.

14. One of the water disinfection methods is based on the oligodynamic action exerted by:

A. Heavy metals.

B. Chlorine.

C. Hydrogen peroxide.

D. Ozone.

E. Pantocid.

15. Various types of water used in pharmaceutical practice are subject to different requirements. Sterility requirements are established for:

A. Purified water.

B. Demineralized water.

C. Water for injections.

16. The storage period for water for injections from the moment of production must not exceed:

A. 6 hours.

B. 12 hours.

C. 24 hours.

D. 48 hours.

17. The storage period for purified water from the moment of its production is no more than:

A. 24 hours.

Б. Two days.

B. Three days.

Г. Four days.

Д. Five days.

18. Various methods are used to detect pyrogens. Indicate the main, officially accepted control method:

A. Biological.

Б. Physical.

B. Chemical.

Г. Microbiological.

19. According to the requirements of the State Pharmacopoeia of Ukraine, the LAL test, based on the interaction of pyrogens with the amoebocyte lysate of which Organism, is used to detect pyrogens (bacterial endotoxins):

A. Crab.

Б. Rabbit.

B. Rat.

Г. Frog.

20. A glucose solution was prepared using pharmacy-prepared water for injection, and its administration to a patient caused a pyrogenic reaction. Indicate its primary symptom.

A. Hypothermia.

Б. Hyperthermia.

B. Hypotension.

Г. Hypertension.

SITUATIONAL TASKS

1. Provide a hygienic Assessment of the quality of centralized or decentralized drinking water supply. When completing this task, relevant regulatory documents should be used — State Sanitary Rules and Norms (DSSRN) "Drinking Water. Hygienic Requirements for the Quality of Centralized Public Drinking Water Supply No. 383 dated 23.12.96" and "Safety Indicators of Water from Decentralized Sources" (See Appendix, Tables 12 and 13).

2. Prepare a written conclusion on the suitability of the water for drinking purposes.

3. Propose (if necessary) measures to improve the quality of drinking water.

Problem 1.

Laboratory Analysis of the water showed: total plate count 150, coliform index 5, odor 4 points, aftertaste 3 points, color 20°, turbidity 0.5 mg/L, total hardness 7 mEq/L, sulfates 450 mg/L, chlorides 240 mg/L, copper 0.8 mg/L, iron 0.2 mg/L, chlorophenols 0.0001 mg/L, total dissolved solids 980 mg/L, total alkalinity 5.8 mg/L, magnesium 21 mg/L, fluorine 0.3 mg/L, aluminum 0.2 mg/L, arsenic 0.005 mg/L, nitrates 41 mg/L, pesticides 0.00009 mg/L, oxidizability 3 mg/L, total alpha activity 0.1 Bq/L, residual free chlorine 0.1 mg/L.

Problem 2.

Laboratory analysis of the water showed: total plate count 95, coliform index 2, odor 5 points, aftertaste 4 points, color 21°, turbidity 1.2 mg/L, total hardness 8 mEq/L, sulfates 250 mg/L, chlorides 22 mg/L, iron 0.3 mg/L, manganese 0.1 mg/L, chlorophenols 0.002 mg/L, total dissolved solids 590 mg/L, magnesium 14 mg/L, fluorine 1.2 mg/L, total alkalinity 5.5 mEq/L, selenium 0.005 mg/L, nickel 0.04 mg/L, fluorine 1.8 mg/L, oxidizability 8 mg/L, total beta activity 0.9 Bq/L, residual free chlorine 0.3 mg/L.

Problem 3.

Laboratory analysis of the water showed: total plate count 70, coliform index 2, odor 5 points, aftertaste 6 points, color 24°, turbidity 0.8 mg/L, total hardness 7 mEq/L, sulfates 450 mg/L, chlorides 210 mg/L, iron 0.9 mg/L, copper 1.0 mg/L, chlorophenols 0.0001 mg/L, manganese 0.08 mg/L, total dissolved solids 980 mg/L, fluorine 0.6 mg/L, total alkalinity 6.8 mEq/L, magnesium 15 mg/L, aluminum 0.4 mg/L, lead 0.01 mg/L, fluorine 1.9 mg/L, trihalomethanes 0.06, pesticides 0.00005 mg/L, oxidizability 3.4 mg/L, total beta activity 0.05 Bq/L, residual free chlorine 0.4 mg.

Problem 4.

Laboratory analysis of the water showed: total plate count 105, coliform index 1, odor 4 points, aftertaste 3 points, turbidity 0.7 mg/L, color 25°, sulfates 490 mg/L, chlorides 480 mg/L, copper 0.8 mg/L, chlorophenols 0.0003 mg/L, total dissolved solids 980 mg/L, total hardness 5.8 mEq/L, pH 7.0, magnesium 50 mg/L, fluorine 1.4 mg/L, aluminum

0.5 mg/L, arsenic 0.005 mg/L, selenium 0.004 mg/L, pesticides 0.00003 mg/L, nitrates 55 mg/L, oxidizability 10 mg/L, total alpha activity 0.03 Bq/L, beta activity 0.9, residual free chlorine 0.3 mg/L.

Problem 5.

Laboratory analysis of the water showed: total plate count 56, coliform index 0, odor 1 point, aftertaste 2 points, turbidity 0.1 mg/L, color 15°, sulfates 500 mg/L, chlorides 150 mg/L, total dissolved solids 800 mg/L, copper 0.05 mg/L, manganese 0.01 mg/L, magnesium 14 mg/L, total hardness 7 mEq/L, chlorophenols 0.0 mg/L, fluorine 0.9 mg/L, pH 8.1, aluminum 0.1 mg/L, lead 0.009 mg/L, nitrates 12 mg/L, pesticides 0.00003 mg/L, oxidizability 2 mg/L, total alpha activity 0.9 Bq/L, beta activity 1.2 Bq/L, residual free chlorine 0.3 mg/L.

Problem 6.

Laboratory analysis of the water showed: total plate count 97, coliform index 1, odor 6 points, aftertaste 6 points, turbidity 0.7 mg/L, color 27°, sulfates 430 mg/L, chlorides 250 mg/L, total dissolved solids 790 mg/L, magnesium 50 mg/L, fluorine 0.8 mg/L, pH 8.0 mEq/L, aluminum 0.9 mg/L, arsenic 0.04 mg/L, selenium 0.01 mg/L, lead 0.02 mg/L, nickel 0.05 mg/L, nitrates 49 mg/L, chloroform 0.08 mg/L, pesticides 0.00003 mg/L, oxidizability 5.8 mg/L, total alpha activity 0.06 Bq/L, beta activity 0.9 mg/L, residual free chlorine 0.45 mg/L.

Problem 7.

Laboratory analysis of the water showed: total plate count 87, coliform index 4, odor 2 points, aftertaste 4 points, turbidity 0.6 mg/L, color 22°, sulfates 770 mg/L, chlorides 240 mg/L, total dissolved solids 1400 mg/L, copper 1.2 mg/L, manganese 0.2 mg/L, iron 0.3 mg/L, total hardness 12 mEq/L, chlorophenols 0.00001 mg/L, magnesium 84 mg/L, fluorine 1.3 mg/L, pH 8.0 mg/L, aluminum 0.1 mg/L, arsenic 0.003 mg/L, lead 0.006 mg/L, fluorine 0.9 mg/L, pesticides 0.00004 mg/L, oxidizability 3.5 mg/L, total alpha activity 0.002 Bq/L, beta activity 0.9 Bq/L, residual free chlorine 0.2 mg/L.

Problem 8.

Laboratory analysis of water from a decentralized source (well) showed: total plate count 4, coliform index 0, odor 1 point, aftertaste 1 point, color 5°, transparency 50 cm, sulfates 50 mg/L, chlorides 5 mg/L, copper 0.4 mg/L, iron 0.1 mg/L, magnesium 0.3 mg/L, fluorine 0.2 mg/L, total dissolved solids 95 mg/L, total hardness 4 mEq/L, nitrates 0.02 mg/L, pH 6.5, hardness 5.0 mEq/L, oxidizability 1 mg/L, residual free chlorine 0.03 mg/L.

Problem 9.

Laboratory analysis of water from a decentralized source (well) showed: total plate count 250, coliform index 8, odor 3 points, aftertaste 3 points, color 4°, transparency 29 cm, sulfates 330 mg/L, chlorides 25 mg/L, fluorine 2.9 mg/L, total hardness 5 mEq/L, total dissolved solids 385 mg/L, ammonia 0.5 mg/L, nitrites 0.004 mg/L, nitrates 49 mg/L, oxidizability 20 mg/L, residual free chlorine 0.03 mg/L.

Problem 10.

Laboratory analysis of water from a decentralized source (well) showed: total plate count 3, coliform index 0, odor 1 point, aftertaste 1 point, color 48°, transparency 35 cm, sulfates 60 mg/L, chlorides 4 mg/L, fluorine 1.4 mg/L, strontium 9 mg/L, total hardness 4 mg/L, total dissolved solids 70 mg/L, ammonia 0.02 mg/L, nitrates 5 mg/L, oxidizability 5 mg/L, residual free chlorine 0.04 mg/L.