“L.I. Medved’s Research Center of Preventive Toxicology, Food and Chemical Safety”, Ministry of Health of Ukraine, Kyiv, Ukraine

Abstract. The objective of this work is to study the toxic properties of disinfectants Forticept, Chisto and Polidez in different routes of the exposure, to carry out a hygienic assessment of working conditions during disinfection activities to resolve the issue of the possibility of using them for their intended purpose.
Methods of the study. Toxicological, hygienic, chemical and analytical, statistical. Results and their discussion. The toxicity parameters of disinfectants Forticept, Chisto and Polidez were determined when affecting laboratory animals in different ways. It was found that the formulation of these agents is less toxic than their active components (salts of polyhexamethyleneguanidine). The studied disinfectants belong to low-hazardsubstances in case of a single epidermal exposure (hazard class 4, GOST 12.1.007-76), moderately dangerous substances when ingested through the respiratory tract and gastrointestinal tract (hazard class 3, GOST 12.1.007-76). Irritant effect on the eye mucous membranes depends on the concentration of the active ingredient and the excipients included in their formulation. In case of repeated epidermal exposure, they do not have an irritating and unfavourable effect on the laboratory animals. The species sensitivity of laboratory animals to the effects of disinfectants was not observed. The results of sanitary and chemical studies showed that in different disinfection regimes, the active ingredient of these agents (PHMG-hydrochloride) was not observed in the air of the premises during and after treatment, after airing the room and on the surfaces (after cleaning with warm water).
Conclusions. Toxic properties are more pronounced in Polidez-50, which contains two active components (PHMG-hydrochloride and PHMG-phosphate) than in agents with one active component — PHMG-hydrochloride (Forticept and Chisto).
- When disinfectants contact the skin, they are classified as low-toxic substances — hazard class 4 according to GOST 12.1007-76 [41].
- Working solutions of Polidez, Forticept and Chisto that are used to disinfect objects are classified as low-risk substances.
- A hygienic assessment of the use of Polidez-50 disinfectant in various disinfection regimes at the recommended rates of consumption showed that the active ingredient was not observed in the air of the premises during treatment and after ventilation, as well as on the treated surfaces.
- Taking into account toxic parameters of Forticept, Chisto and Polidez in case of different routes of exposure, as well as the results of sanitary and chemical studies, it is possible to use them for target purpose as agents that provide bactericidal, fungicidal, tuberculosis and virucidal actions for disinfection of premises and surfaces (including floors, walls, dishes, etc.), and antiseptic treatment of open skin areas.
Key words: toxic properties, disinfectants, polyguanidines, Forticept, Chisto, Polidez.

Relevance. Throughout history, mankind has faced infectious diseases [1–6]. In addition to a number of necessary measures, the use of effective antiseptics — disinfectants — contributes to the normalisation of the situation, localisation of an epidemiologically unfavourable place, and a decrease of the risk of spreading of infectious disease foci. The assortment of disinfectants is quite large with the prevalence of traditional biocidal agents: chloroactive, oxygen-containing, quaternary ammonium compounds, as well as compounds containing salts of heavy metals (copper, stannum, etc.) [7–10].     Although chloroactive compounds inhibit most microorganisms, they are not effective enough or completely ineffective in respect of spore forms (bacilli), viruses, Pseudomonas aeruginosa, protozoan cysts; and oxygen-containing compounds are characterized by a lower efficiency. Most of the chemical compounds of these classes are very aggressive and toxic, so using them poses a certain threat to human health, is unsafe for the environment, causes corrosion of equipment, damages and discolours materials. Thus, chlorine-containing disinfectants have a pronounced irritant effect on the skin, mucous membranes of the eyes and upper respiratory tract. They harm the environment since they are the main sources of dioxins. Compounds containing heavy metals that are commonly used in wood preservatives and non-fouling paints are highly toxic and environmentally unsafe: They easily enter the human and animal body through the food chain causing serious consequences [7–10].

The most promising disinfectants include high-molecular biocidal agents based on the derivative of the guanidine nitrogenous base — polyalkylene guanidine (PAG) and its high-molecular salts [11–14].

The empirical formula of PAG: (-С₇Н₁₆N₃-)_n.

The structural formula of PAG

where n and x are usually equal to 4–50, y = 1–2.

The PAG mechanism of action on bacteria is based on their ability to change the properties of the cell membrane of the microorganism. PAG high-molecular salts are solids soluble in water, which have the properties of a cationic polyelectrolyte and a strong organic base. After dissociation of PAG salts, the formed cations react with membranes of bacteria that have a negative charge. In this case, the lipophilic groups of the agent facilitate the disaggregation of the lipoprotein membrane of bacteria, as a result of which there is a disturbance of the osmotic balance, loss of potassium and phosphorus from the bacterial cell. Under the action of the agent, the bacterial cytoplasmic membrane breaks down and its osmotic balance is disturbed, which results in the death of the bacterium.

Disinfectants based on PAG and its salts have a wide range of biocidal action: They equally affect the aerobic and anaerobic microflora, are effective against Pseudomonas aeruginosa, mycobacterium tuberculosis, suppress pathogens of some particularly dangerous infections (legionellosis, glanders, plague), have a virucidal effect regarding pathogens of poliomyelitis, HIV, hepatitis, herpes, influenza.

It was experimentally shown that PAG and its salts are absorbed through intact skin, but due to the low oil/water distribution coefficient, the rate of transepidermal resorption of the polymer through intact skin is much lower than of its analogue, chlorhexidine bigluconate. Drying on the skin surface, the polymer forms a film that prevents further resorption of the antiseptic, so the absorption of the polymer upon contact with the skin quickly ceases. Apparently, a decrease in the toxicity of PAGs is due to the different permeability of the skin for the polymer, which affects its impassability through cell membranes and low mobility of large molecules [12–14].

It was found that the toxicity of PAG depends to a large extent on the chemical nature of the A- anion. Thus, as the electronegativity of A- anion decreases, LD₅₀ value also decreases.  The anion effect is due to its donor¬acceptor properties: Anion acceptors (for example, Cl-, OH-) pull electrons onto themselves increasing the positive charge on the guanidinium cation. Redistribution of electron density in the guanidine group creates tension in the entire macromolecule: It takes an elongated conformation, in which all the guanidine groups are readily available and implement their reactivity when interacting with the cell. Anions of phosphoric, gluconic and other organic acids, on the contrary, give the electron density to the guanidine group reducing its positive charge. The anion-initiated redistribution of the electron density in the guanidine group is spread along the polymer chain and enhances the intramolecular interactions of the functional groups removed in the chain order. As a result, the macromolecule takes a spiral conformation, which is stabilised by hydrogen bonds and van der Waals interaction of hexamethylene fragments (the anion determines the step of the intramolecular spiral). In this case, part of the guanidine groups is blocked and loses its reactivity, in addition, the complementarity of the polyguanidine macromolecule and receptors of the cell membrane structures may be impaired.

It has been found that polyhexamethylene guanidine (PHMG) and its salts are more effective than PAG regarding many bacteria and fungi.

The PHMG macromolecule is a well-balanced system in which the guanidine group has a positive charge and provides bactericidal properties; A- anion effects the degree of delocalisation of the positive charge and thus affects the toxicity; the hexamethylene chain promotes the redistribution of the electron density in the macromolecule and, in addition, regulates the hydrophilic and¬hydrophobic balance of the molecule. The enhancement of the polymer biocidal action effectiveness in comparison with the low molecular weight analogue (chlorhexidine bigluconate) is provided by the cooperative interaction of many chemically bound guanidine groups with a microbial cell, and the decrease of toxicity is provided by the possibility of the electron density delocalisation along the polymer chain and its conformational transformations.

It was found that with an increase in the molecular mass of PHMG, its minimum retentive concentration decreases reaching a minimum value at MM = 10,000 Da. [15–17]. Further increase in the size of PHMG macromolecule is not accompanied by a significant increase of its antimicrobial activity; Japanese authors noted its growth to MM ~ 80,000 Da [28]. The macromolecular nature of PHMG provides a prolonged antimicrobial effect of the agent: Unlike low-molecular compounds, antimicrobial effect of which lasts only a few hours (at best, several days), the polymer forms a biocidal film on the surface, which provides a long-term (several months) protection of the treated surface from microorganisms. Due to the polymeric nature, PHMG and its salts have cationic surface-active properties.

According to the literature, PHMG belongs to low-risk substances when applied to the skin: LD₅₀ >10,000 mg/kg [17]. PHMG low toxicity is explained by the fact that warm-blooded animals have enzyme systems in their body capable of causing degradation of guanidine-containing polymers. Therefore, the “critical security factor” (CSF-factor) that shows what fold this agent is toxic for pathogenic microflora than for humans is more than 3,667 regarding various salts of PHMG.

The information search showed that a number of new biocidal agents (more than 24 % of the total assortment of disinfectants) were made based on PHMG [18–26].

According to the literature data, most disinfectants, the active component of which is the salts of PHMG, do not cause irritation of the skin and eye mucous membranes. Regarding oral toxicity, they are classified as moderately hazardous compounds: LD₅₀ for mice is 450–500 mg/kg, for rats — 815–3,200mg/kg, for guinea pigs — 750–900 mg/kg. They have moderately pronounced cumulative properties: C_cum (cumulative coefficient) = 2.9–3.2. The median lethal dose of various PHMG effect on the body through the dermal contact is 10,000–15,000 mg/kg. The median lethal concentration of substances when administered through the respiratory tract is 4 to 20 mg/L for rats.

When administering these agents to laboratory animals, a general toxic effect is observed. The threshold of the irritant action and general toxicity of these agents is between 190 mg/kg and 25 mg/kg [15–17]. Agents may have a mild allergenic effect [29–30].
Mutagenic and carcinogenic properties of these disinfectants were not observed. The study of PHMG effect on embryogenesis of white rats at oral exposure detected an increase in pre-implant death, total embryonic mortality, which is due to the general toxic effect on the pregnant female. The non-effective dose for the mother is 0.1 mg/kg, and 1.0 mg/kg — for the foetus.

Metabolism of salts of polyhexamethylenguanidine (PHMG-phosphate, PHMG-hydrochloride) causes a replacement of chloride or phosphate anion with gluconate anion in the body of laboratory animals; further, there is a hydrolysis of guanidine groups with their transformation into urea ones, as well as the destruction of polymer chains into separate fragments.    The information search did not detect any information about the studies on determining the content of PHMG salts in environmental objects when sanitising with disinfectants. There is information that PHMG salts may have adverse effects on the human body [30].

According to the Law of Ukraine No. 4004-XII “On the Provision for Sanitary and Epidemic Comfort of the Population” dated February 24, 1994, the assortment of disinfectants is constantly expanding. The need to develop new disinfectants is due to the fact that the microbial background itself is constantly changing, adapting to traditional disinfectants. Therefore, studies on the toxicological and hygienic assessment of new effective disinfectants are relevant.

Domestic disinfectants Forticept, Chisto and Polidez are offered for registration in Ukraine. The active ingredient of these agents is PHMG salts (PHMG-hydrochloride, PHMG-phosphate).

The empirical formula of PHMG-hydrochloride (-С₇Н₁₆N₃Cl)_n. Structural formula

The empirical formula of PHMG-phosphate (-С₇Н₁₆N₃ H₃PO₄)_n. Structural formula

Mm of salts is < 6,000

Disinfectants Forticept, Chisto and Polidez have bactericidal (against Staphylococcus aureus, Pseudomonas aeruginosa Escherichia coli, MRSA 1, Bacillus subtilis), fungicidal (Candida albicans Trichophyton mentagrophytes Aspergillus niger), tuberculocidal and virucidal (against influenza A, bird influenza viruses) action.

Forticept and Chisto are recommended for the antiseptic treatment and disinfection of the human skin surface. It is recommended to use them for the prevention of infections in case of superficial cutaneous lesions, including grazes, scratches, cracks, for the treatment of the following items: personnel hands skin, health and sports institutions of various profiles, employees of children’s and social protection institutions, pharmacies, food, cosmetic, and pharmaceutical enterprises, microbiological industry, transport, communal facilities and other facilities, the activity of which requires compliance with hygiene rules and regulations. They can also be used for the antiseptic treatment of hands before surgical procedures, blood sampling, injections, punctures, vaccinations, and after cosmetic procedures (piercing, tattoo, etc.).    Polidez is used for disinfection of the aerial environment in premises with epidemiological problems (foci of droplet or intestinal infections), disinfection of surfaces, furniture, medical disposable and reusable equipment, medical devices, apparatus, and patient-care items.

The brief literature review indicates the need to study the toxicological properties of disinfectants based on PHMG salts, as the information on the parameters of their toxicity and the degree of danger was absent.

The active ingredient of Forticept is PHMG-chloride, which is contained in Chisto in a smaller amount (4-fold). Polidez is available in 4 trademarks: Polidez-2, Polidez-20, Polidez-30 and Polidez-50. The active ingredient of the agent is a mixture of PHMG salts: PHMG-hydrochloride and PHMG-phosphate in various proportions. The composition also includes auxiliary components: panthenol, ethyl alcohol, glycerine, hydroxyethyl cellulose and purified water.

According to the requirements of the legislation, new substances that in a prospect will be used in the national economy must be subject to tests regarding the parameters of the toxicity and the degree of their danger.

The purpose of this work was to conduct studies on the toxic properties of Forticept, Chisto and Polidez at different routes of exposure, to perform work hygienic evaluation when conducting disinfection using these agents to decide if it is possible to use them for the intended purpose.

Objects and methods of the study. The objects of the study included Wistar Han white rats, mice, Chinchilla rabbits, guinea pigs; disinfection agents Forticept, Chisto, Polidez. The study was carried out in accordance with the requirements of the quality control system of the State Enterprise “L. I. Medved’s Research Center of Preventive Toxicology, Food and Chemical Safety” of the Ministry of Health of Ukraine, which takes into account the requirements of ISO/ES 17025 (accreditation certificate No. 2H375 dated May 22, 2015). Toxicological studies were performed in accordance with the Assessment of Toxicity and Hazards of Disinfectants Methodology Guidelines, Assessment of Toxicity and Danger of Disinfectants Methodology Recommendations and the requirements set forth in Toxicometry of Toxic Substances that Contaminate the Environment [32–33]. The study of the agent percutaneous action in the settings of the subchronic experiment was carried out in accordance with the Assessment of the Effect of Hazardous Chemical Compounds on the Skin and Substantiation of the Maximum Permissible Levels of Skin Contamination Methodology Guidelines [35]. The criterion of the effect of disinfectants at different routes of exposure was the manifestation of intoxication symptoms in animals and their mortality. Follow-up control determined integral indicators of toxic effect: behaviour, feed and water intake, body weight, haematological parameters of peripheral blood of rats. After the end of the studies, a macroscopic examination of the internal organs was carried out. The median lethal dose (LD₅₀) of agents was determined in case of oral and dermal exposure, median lethal concentration (LC₅₀) in case of intake through the respiratory tract, local irritant properties when applied to the skin and eye mucous membranes [36]. The sensitisation properties of the agents were studied on guinea pigs according to the method of O. G. Alekseieva and A. I. Petkevych, the immunological indicators were also determined: Mast cell degranulation reaction according to Schwartz and specific leukocyte lysis reaction [36, 37]. The cumulative properties of disinfectants were studied when administered orally in white rats within 2 months. The maintenance, care and all manipulations with laboratory animals were carried out in accordance with the provision of the General Ethical Principles for Experiments on Animals and DSTU IS0 10994-2:2004 [40]. Studies on the assessment of working conditions when using disinfectants were carried out based on determining the content of the active ingredient (PHMG-hydrochloride) in the aerial environment, as well as in wipe-samples from the treated surfaces. Quantitative determination of PHMG-hydrochloride was carried out in accordance with the Performing of Measurement of the Mass Concentration of Polyhexamethylene Guanidine Salts in the Air of Working Place by Spectrophotometric Method Methodological Guidelines. The method is based on measuring of the compound solution (at an optical density of 540 nm), which is formed by the interaction of PHMG salts with p-nitrophenyldiazonium in alkaline medium. The lower limit of measurement in the air is 0.5 mg/m3. The air was aspirated to the АФА-ВП-10 filter with the rate of 5–10 L/min. Fifty litres of air were taken for the study.

The results were processed statistically with the help of a microcomputer, with the determining of the Student’s test and likelihood indicators [41].

Results and their discussion. When Forticept was orally administered in white rats, a decrease in motor activity, rapid shallow breath were observed. During the experiment (14 days), experimental animals presented with a significant decrease in body weight gain in comparison with control animals. Thus, a week after administration, the body weight of the experimental males increased by only 1.5 %, while in the control group a 4-fold increase was observed, and in two weeks it was 1.5-fold. In experimental females, a body weight gain in a week after exposure to the agent was not observed. Compared to the initial value, their body weight decreased by 0.4 %, while body weight in the control females increased by 3.5 % during this period. Two weeks later, the body weight gain of the experimental group of female rats was 6 times less than that of the control group. The results of the studies showed that the female rats appeared to be more sensitive to the effect of the agent. LD₅₀ for female rats is 2,920 mg/kg, for male rats — 4,700 mg/kg. After intragastric administration of Chisto, the symptoms of intoxication in male and female rats were similar and manifested as an impairment of the nervous system. During the follow-up period, females did not gain any weight. The body weight gain in male rats was at the control level. LD50 of Chisto for rats (males and females) is 5,000 mg/kg. Compared to Forticept and Chisto, a single administration of Polidez-50 through the gastrointestinal canal caused a more pronounced pattern of intoxication in rats (males and females) and mice (males and females). It manifested itself as the inhibition of motor activity, impaired coordination of movements, shallow breathing, weak reaction to tactile and noxious stimuli, vacualisation. Tonic-clonic muscle activity was observed in the terminal stage. Within 3 days, the animal death and a decrease in the body weight gain in comparison with the initial body weight were observed. Thus, a week after the agent administration, the body weight of the experimental females decreased by 6.3–6.9 % in comparison with the initial weight. Two weeks later, they gained some weight, but it was still lower than the initial value by 4.9–5.5 %, while in the control group, the body weight gain of females increased by 3.5 % and 7.2 %, respectively. Macroscopic examination detected liver plethora, spleen induration, bloating and flatulence. LD₅₀ of Polidez-50 is 1,300 mg/kg for white rats (males and females) and 1,025 mg/kg for mice (males and females). There are no specific features observed regarding the sensitivity of animals to the action of disinfectants depending on the type and sex of animals. The percutaneous action of Forticept and Chisto was observed when applied to the skin of female rats. There was a significant decrease in the body weight of the experimental animals by 10.9 % after a single application of Forticept, and by 5.3 % after application of Chisto. The macroscopic studies of rats carried out at the end of the experiment showed that the condition of their internal organs did not differ from those of the control group. LD₅₀ of Forticept and Chisto for white rats is 4,000 mg/kg.

In case of a single application of a disinfectant Polidez-50 at a dose of 4,000 mg/kg to the skin, laboratory animals showed the symptoms of intoxication similar to those observed at oral exposure. Lethal outcomes (33.3 %) were observed in rabbits. Body weight gain in rats and rabbits was decreased.

The toxic effect was observed in case of inhalation exposure of laboratory animals to Forticept, Chisto, and Polidez. Male and female rats, which were exposed to a single dose of saturated steam of the agent at a concentration of 20,000 mg/m³ through the respiratory organs (1 hour exposure), showed similar symptoms of intoxication: shallow breathing, sanious discharge from the nasal sinuses, impaired coordination of movement, decreased motor activity. The condition of the animals returned to normal within 4–5 hours after exposure. There were no lethal outcomes in rats. After inhalation, the experimental females gained the body weight slightly lower compared to the control animals. A week after exposure to Polidez-50, the females of the experimental group increased their body weight by 1.5 % (control group — 3.9%), two weeks after — by 3.4 %, while the control group by 5.6 %.  In the males of the experimental group, the body weight gain throughout the entire experiment was 2–2.5 times lower than of those from the control group. In case of exposure to Forticept and Chisto through the respiratory tract, intoxication was less pronounced, the body weight gain in the experimental and control groups (males and females) was at the same level. Based on the obtained data, it can be concluded that LC50  of Polidez, Forticept and Chisto for male and female rats is more than 20,000 mg/m³. Application of Forticept, Chisto and Polidez-20 to the skin in the amount of 20 mg/cm² do not cause changes in the skin in laboratory animals. The body weight gain in the experimental group was at the control level.  When Polidez-50 was applied to the skin of rabbits and rats in the amount of 20 mg/cm², skin irritation was observed: erythema, skin flaking, a coat had a pale yellow colour at the edge of the treated area. Total irritation index is 2 points.

After intraconjunctival administration of native Forticept and Chisto to rabbits, a mild lacrimation was observed. Three–four hours after the exposure, the eye was closed. In a day, there was conjunctival hyperaemia (2 points) and a small amount of seropurulent discharge (2–1 points). During the entire period of observation, there was no palpebral oedema and changes of tactile sensitivity. Polidez-20 irritant effect on the mucous membranes was manifested as diffuse hyperaemia and mild conjunctival oedema, seropurulent discharges (total irritant effect at the level of 5 points). These events passed in 7 days after the exposure. More pronounced changes were observed under the influence of Polidez-50. The noted changes due to this compound passed in 14 days after the exposure.

There were no manifestations of the eye mucous membrane irritation after exposure to 1.0–2.0 % of working solutions of disinfectants, which are recommended for disinfection of objects.

Thus, the conducted studies showed that the degree of irritant properties depends on the concentration of the active ingredient. The irritant effect of these agents is associated with a higher amount of active ingredients that are contained in the formulation and the presence of various auxiliary components.

Forticept, Chisto and Polidez-20 do not cause specific sensitisation. During the skin application with these agents in a native form, 50 % and 25 % aqueous solution, the skin of guinea pigs was clean and of normal colour throughout the experiment. The skin reaction to the effect of the agents was 0 points in all experimental animals. 50 % aqueous solution of the agents was used as a sensitising concentration, and the native preparation was used as the testing agent. Study of the sensitising properties of the agents showed that after intradermal administration of Polidez-20, Forticept or Chisto (200 μg) to the guinea pig’s ear, no local skin reaction was observed. The application of agents in the sensitising concentration within 7 days also did not cause irritant effect. Throughout the experiment, the animals’ skin was clean, of a normal colour. In 1, 3, 6, 24 and 48 hours after the first and second testing, the skin reaction to antigen exposure in experimental and control guinea pigs was 0 points. The results of the conducted studies allow to draw a conclusion that these agents belong to substances which do not have sensitisation properties. At the same time, Polidez-50 showed mild allergenic properties. There were a presence of hyperaemia and a change in immunological indicators (mast cell degranulation reaction according to Schwartz and specific leukocyte lysis reaction) observed.

The symptoms of intoxication or lethal outcomes in rats were not observed within 2 months in case of oral administration of Forticept, Chisto and Polidez-20. The body weight gain in animals of the experimental group was at the control level. The obtained results indicate that these agents do not have cumulative properties. C.cum. > 5.

The effect of Forticept and Chisto on the skin was also performed under conditions of a subchronic experiment in female rats, which are the most sensitive to the effects of the agents. The application of the agents was carried out daily 5 times a week within a month in a dose of 200 mg/kg, which was 1/20 of the epidermal LD₅₀. The symptoms of intoxication or lethal outcomes in animals were not observed. There were no signs of any skin irritations: erythema, oedema, skin flaking and infiltration. There were no pain in the treated area, an increase in the thickness of the skin fold, or other clinical signs of damaging action observed on palpation. Tactile sensitivity was normal. Further, the treated skin areas were evenly covered with a coat, like in the control group. Behaviour and feeding of animals from the experimental group were at the control level. The body weight gain of the experimental and control animals was at the same level throughout the experiment. A slightly greater body weight gain by females was noted when applying Chisto. Macroscopic studies showed that the appearance of the experimental females (their coat looked neat, the skin was without flaking, the mucous membranes of the eyes and nose were without irritations and discharges) and the condition of their internal organs did not differ from those of the control group: No expanding and filling with the blood of brain mater vessels, haemorrhages and other abnormalities were observed. The lungs were pink, filled with air, elastic. There was no foreign matter in the chest cavity. No changes in the thymus and heart were observed. Oesophagus, trachea and thyroid gland were within normal limits. No abdominal adhesions and foreign matter in the abdominal cavity were observed. The surface of the liver, kidneys, and spleen was smooth and shiny. In section, the tissue of the liver and spleen was homogeneous in colour and consistency. Kidneys and adrenal glands are of normal size and shape, without haemorrhage. The pancreas has a distinctive colour, a homogeneous consistency, without necrosis. The external and internal mucosa of the stomach and intestine is within normal limits. Uterus and ovaries are within normal limits. The performed studies indicate that Forticept and Chisto do not have unfavourable effect in rats in case of repeated application to the skin of the animals.

During disinfection activities, an inhalation intake of the active ingredient (PHMG-hydrochloride) of a disinfectant into the human body is possible. Taking into account the possibility of inhalation intake of PHMG-hydrochloride during disinfection, a series of experiments was conducted to study the dynamics of its content in the aerial environment. To do this, disinfection of an area of 11.6 m² (floor, ceiling, walls, doors, furniture) in a room that simulates living conditions (area is 18 m², height is 2.7 m, air temperature is 18^oC) was conducted with 5 % solution of Polidez-50 in different ways:

- Irrigation with a consumption rate of 200 mL/m² and exposure of 90 min. Air samples were collected in the breathing zone, at a height of 1 m and 0.5 m from the treated surface;

- Wiping with a consumption rate of 150 mL/m² and exposure of 90 min. Air samples were collected in the breathing zone, at a height of 1 m and 0.5 m from the treated surface and after 15 minutes of ventilation.

- Soaking of medical devices in a desiccator with a volume of 3.0 L, d=18 cm. At the end of the exposure (90 minutes), air samples were taken at a distance of 15 cm, 2.0 m, 3.0 m from the location of the desiccator with medical devices. After ventilation (30 minutes) and washing the medical equipment treated with Polidez-50 with warm water for 3 minutes, air samples were taken at a distance of 15 cm, 2 m and 3 m from the location of the desiccator.

- Soaking dishes with food residues (conditionally contaminated with parenteral viral infections), in a 3.0 L desiccator, d=18 cm, according to the recommendations stipulated in the Methodology Guidelines. At the end of the exposure (90 minutes), air samples were taken at a distance of 15 cm, 2.0 m, 3.0 m from the desiccator and 30 minutes after ventilating the room and washing the disinfected dishes with warm water for 3 minutes.

The results of the conducted studies showed that the content of PHMG-hydrochloride in the aerial environment of the room and on the surfaces (after washing with warm water) did not exceed the hygienic standards in none of the studied samples when using the disinfectant Polidez-50: MAC in the working zone area is 2.0 mg/m³, MAC in reservoir water of household and amenity use is 0.1 mg/L.

Thus, the results of sanitary and chemical studies showed that when disinfecting by irrigation, wiping and soaking, PHMG-hydrochloride was not observed (at the sensitivity level of the analytical method of determination) in the air of the premises during and after treatment, after airing the room and on the surfaces (after cleaning with warm water).

 

CONCLUSIONS

- The conducted studies showed that toxic properties are more pronounced in Polidez-50, which contains two active components (PHMG-hydrochloride and PHMG-phosphate) than in agents with one active component — PHMG-hydrochloride (Forticept and Chisto).

- The toxicity of the studied disinfectants depends on the amount of active ingredient. The least dangerous agents are Polidez-20 that contains 2.5 times less of PHMG-hydrochloride and PHMG-phosphate than Polidez-50, and Chisto that contains 2.5 times less of PHMG-hydrochloride than Forticept.

- When disinfectants contact the skin, they are classified as low-toxic substances — hazard class 4 according to GOST 12.1007-76 [41].

- Working solutions of Polidez, Forticept and Chisto, which disinfect objects, are classified as low-risk substances.

- A hygienic assessment of the use of Polidez-50 disinfectant in various disinfection regimes (wiping, irrigation, soaking) at the recommended rates of consumption showed that the active ingredient (PHMG-hydrochloride) was not observed in the air of the premises during treatment and after ventilation, as well as on the treated surfaces (after washing with warm water for 3 minutes).

- Taking into account toxic parameters of Forticept, Chisto and Polidez at different routes of exposure, as well as the results of sanitary and chemical studies, it is possible to use them for target purpose as agents that provide bactericidal, fungicidal, tuberculosis and virucidal actions for disinfection of premises and surfaces (including floors, walls, dishes, etc.), and antiseptic treatment of open skin areas.

The results of the research were used when considering the registration of Forticept, Chisto and Polidez in Ukraine for the use as disinfectants according to the intended purpose.

 

REFERENCES

1. Priority directions of hygiene science development in Ukraine / Yu. I. Kundiiev, Ie. H. Honcharuk, Yu. S. Kahan [et al.] // Priority problems of hygienic science, medical ecology, sanitary practice and health care. – K.: MoH of Ukraine, 1995.– P.1. – P. 13-17.

2. Surmasheva O. V. Infectious Disease Incidence in Ukraine / O. V. Surmasheva, E. P. Bernasavska, M. O. Rosada // Dovkillia ta zdorovia. – September. – 2003. – P. 49–52.

3. Onyshchenko G. G. On the state of surveillance for non-specific prevention of infectious diseases and tasks for its improvement / G. G. Onyshchenko // Hihiiena i sanitariia. – 2007. – No. 1. – P.4–11.

4. The incidence of infectious diseases and its connection with environmental pollution / R. A. Sazhok // Priority problems of hygienic science, medical ecology, sanitary practice and health care. – K.: MoH of Ukraine, 1995.– P.1. – P. 120-21.

5. Shkarin V. V. Disinfection. Disinfectation. Deratization / V. V. Shkarin – Nizhny Novgorod: NSMA, 2006. – P.213–260.

6. Epidemiology: textbook [eds. K. M. Syniak]. – Zdorovia. – Kyiv. – 1993. – 460 p.

7. Vietkina I. F. A modern approach to the selection of disinfectants in the system of prevention of hospital-acquired infection (HAI) / I. F. Vietkina, L. V. Komarinskaia [et al.] // FARMindeks Praktik. – 2005. – Ed. 7. – P.13–20.

8. Shandala M. G. Actual issues of general disinfectology / M. G. Shandala – Selected lectures. – M.: Meditsina, 2009 – 111 p.

9. Ukraintsev A. D. A comparative analysis of agents used for the disinfection of dangerous microorganisms / A. D. Ukraintsev, I. G. Vlasov, T. K. Krashennikova [et al.] // Khimicheskaia i biolohicheskia bezopasnost. – 2005. – No. 6. – P. 3-25.

10. Modern means of disinfection and disinfectation. Characteristics, purpose, prospects / [L. S. Fedorova, L. I. Arefieva, L. S. Putintseva, N. A. Veromkovych] – M.: NPO Soiuzmedinform. – 1991. – 38 p.

11. Polyhexamethylene guanidine hydrochloride-based disinfectant: a novel tool to fight meticillin-resistant Staphylococcus aureus and nosocomial infections / Mathias K. Oule, Richard Azinwi, Anne-Marie Bernier [et al.] // Journal of Medical Microbiology. – 2008. – V. 57. – P.1523 – 1528.

12. Assessment of the antifungal activities of polyhexamethylene-guanidine hydrochloride (PHMGH)-based disinfectant against fungi isolated from papaya (Carica papaya L.) fruit / Rose Koffi-Nevry, Ama Lethicia Manizan, Kablan Tano [et al.] // African Journal of Microbiology Research. – 2011.– V. No. 5(24). – P. 4162 –4169.

13. Literature review - efficacy of various disinfectants against Legionella in water systems / B.R.Kim, J.E. Anderson, S.A.Mueller [et al.] // Water Research. – 2002. – V. No. 36. – Р. 4433 – 4444.

14. Evaluation of Antimicrobial Activity of Surface Disinfectants by Quantitative Suspension Method / K. Prasanthi, D.S.Murty, Nirmal Kumar Saxena // International Journal of Research in Biological Sciences. – 2012. – V. No. 2(3). – Р. 124 –127.

15. http://blog.pgmg.ru/post_1230297793.html / I. I. Vointseva, N. A. Polikarpov. Polyalkylene guanidines – biocidal properties and toxicity.

16. http://blog.pgmg.ru/post_1230265539.html /    K.M. Iefymov. Polyguanidines – ecologically safe biocides.

17. Iefymov K. M. Polyguanidines – a class of low-toxic, long-acting disinfectants / K. M. Iefymov, P. A. Gembytskii, A. G. Snezhko // Dezinfektsionnoe delo. – 2000. – No. 4. – P. 28     – 32.

18. Mariievskyi V. F. Polyhexamethylene guanidine hydrochloride: promising biocide agent / V. F. Mariievskyi, S. P. Voronin, I. S. Chekman, A. I. Hrebelnyk // Farmakolohiia ta likarska toksykolohiia. – 2014. – No. 1 (37). – P.17 – 21.

18. http://blog.pgmg.ru/post_1230297793.htm / S. S. Kozak, A. I. Dytiuk, Y. V. Ratkevych. Polyguanidines as promising means for disinfecting and obtaining safe poultry products.

19. Antimicrobial activity, disinfecting properties and toxicity of alkylene (C12-C14) dimethyl benzyl ammonium chloride / L. S. Fedorova, L. I. Arefieva, G. P. Pankratova [et al.] // Sovremennye metody i sredstva dezinfektsii i sterilizatsii. – М., 1989. – P.17.

20. Rutela W. A. Disinfection, sterilization and waste disposal / W. A. Rutela / Hospital-acquired infections / translated from English, eds. R. P. Ventsel. – М., Meditsina. – 2000. – 144 p.

21. Sokolova N. F. Actual problems of non-specific prevention of anthrax / N. F. Sokolova // Theoretical and practical aspects of modern epidemiology / Material of the scientific and practical conference dedicated to the 75th anniversary of the birth of the honoured worker of science of the Russian Federation, academician of the Russian Academy of Medical Sciences Benyamin Lazarevich Cherkassky. – M.: 2009. – P. 59 – 68.

22. Ways to create effective and safe antimicrobial liquid agents and the evolution of public perception of disinfection measures / V. M. Bakhir, B. I. Leonov, S. A. Panycheva [et al.] // Dezinfektsionnoe delo. – 2004. – No. 3. – P. 44 – 49. 

23. New polyguanidines – innovative disinfectants of prolonged antimicrobial action / K. М.Iefymov, A. I. Dytiuk, G. P.Pankratova [et al.] //  Dezinfektsionnoe delo. – 2015. – No. 3. – P. 13–19.

24. Gembytskyi P. A. Polymer biocidal preparation polyhexamethylene guanidine / P. A. Gembytskyi, I. I. Vointseva // Zaporizhzhia: Polihraf, 1998. – P. 43.

25. Abramova I. M., Sukiasian A. N., Kopylova A. I. Sporocidal activity of glutaric aldehyde depending on pH and temperature of solutions // Disinfection and sterilization issues. Collection of scientific papers. – М., 1986. – P. 58–61.

26. Richads J. Withdrawal of Disinfektant Hit by Safety Fears / J.Richads // BBC News on Line: Health. – January 22. – 2002. – Р. 7–9.

27. Bekhalo V. A. Immunobiological features of bacterial cells of medical biofilms/ V. A. Bekhalo, V. M. Bondarenko, Ie. V. Sysoliatina, Ie. V. Nahurskaia// Microbiolohiia. – 2010. – No. 4 (97) – P. 105.

28. The In Vitro Efficacy Testing Of Skin Disinfectants Against Nosocomial Pathogens / S.Jayakumar, M.Kanagavalli, A.S. Shameem Banu [et al.] // Journal of Clinical and Diagnostic Research. – 2011. – V. No. 5(2). – Р. 231 – 235.

29. http://www.antibiotic.ru/cmac/pdf/cmac.2013.t15.n4.p279.pdf

30. http://www.piluli.kharkov.ua/drugs/drug/chlorhexidine-bigluconate/#pobochnie-deistviya

31. http://polyguanidines.ru/sdez/guanidin.htm

32. Assessment of Toxicity and Hazards of Disinfectants Methodology Guidelines. – M. – 2002. – 35 p.

33. Assessment of Toxicity and Danger of Disinfectants Methodology Recommendations.– M. – 1990. – 26 p.

34. Toxicometry of Toxic Substances that Contaminate the Environment. Centre for International Projects of the State Committee for Science and Technology. – M. – 1986. – 426 p.

35. Assessment of the Effect of Hazardous Chemical Compounds on the Skin and Substantiation of the Maximum Permissible Levels of Skin Contamination Methodology Guidelines. – M. – 1989. – 25 p.

36. Methodology guidelines regarding formulation of studies on the irritant properties and the substantiation of the maximum permissible concentrations of selective substances in the working zone air. – M. – 1989. – P.17.

37. Industrial allergens / [L. A. Duieva, V. Iu. Kohan, S. V. Suvorov, R. Ia. Shterenharts] – M.: Centre for International Projects of the State Committee for Nature Protection of the USSR, 1989. – P. 37–53.

38. Study on the Hygienic Regulation of Industrial Allergens in the Working Zone Air Methodology Guidelines. – 1988. – 26 p.

39. DSTU IS0 10994-2:2004 Biological Assessment of Medicinal Products. Part 2. Requirements regarding animal care. – K.: State Committee for Technical Regulation and Consumer Policy of Ukraine, 2006. – 160 P.

40. Ivanov Iu. I. Statistical processing of the results of medical and biological studies using microcalculators according to the programs / Iu. I. Ivanov, O. N. Pohoreliuk – M.: Meditsina. – 1990. – 217 P.

41. GOST 12.1.007-76 Harmful substances. Classification and general requirements.

 

Надійшла до редакції 4.12.17