Email this article Virucidal efficacy of compositions based on lanthanum nitrate hexahydrate
- Authors: Berezhnaya E.V.1, Pashkin I.I.2, Bochkova M.S.1, Kondrashina N.G.1, Kuzmicheva G.M.2, Domoroshchina E.N.2, Nosik D.N.1
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Affiliations:
- National Research Center of Epidemiology and Microbiology named after honorary academician N.F. Gamaleya
- MIREA — Russian Technological University
- Issue: Vol 103, No 3 (2026)
- Pages: 379-385
- Section: ORIGINAL RESEARCHES
- URL: https://microbiol.crie.ru/jour/article/view/19208
- DOI: https://doi.org/10.36233/0372-9311-780
- EDN: https://elibrary.ru/ZLBKXL
- ID: 19208
Cite item
Abstract
Background. The most urgent problem of medicine is the fight against viral diseases. Preventive measures in the form of antiseptics with virucidal effect (VE) play an important role. Antimicrobial properties have been noted in compounds based on rare-earth metals.
The aim of the study was to investigate the VE of lanthanum nitrate hexahydrate (LNH) and compositions based on it in order to establish the possibility of using them in the composition of topical antiseptics.
Materials and methods. The VE of compounds was studied on the model of viruses: poliovirus, adenovirus, human immunodeficiency virus (HIV-1), herpes simplex virus (HSV-1). The VE was assessed by degree of reduction of virus titer in lg TCID50. The following compounds were studied: organic lanthanum composition (OLC), consisting of LNH (La(NO3)3·6H2O), triethylene glycol (TEG), ethyl carbitol (EC), glycerol, H2O and NaOH, as well as solutions of LNH in H2O and glycols: TEG, EC, 1,2-propylene glycol (PG), 1,3-butylene glycol (BG), polyethylene glycol-400 (PEG) and glycols themselves.
Results. OLC reduced titers of poliovirus and adenovirus by 4.0-4.5 lg TCID50 in vitro, on surfaces by 4.0–5.0 lg TCID50 with disinfection time of 1–2 minutes. Components included in OLC reduced the titer of poliovirus by 3.1-4.0 lg TCID50. The reduction of HSV-1 titer of all compounds amounted to 4.0–4.3 lg TCID50. La(NO3)3·6H2O in PG, BG, PEG-400 and EC reduced HIV-1 titer by 4.8–5.3 lg TCID50 at pH below 4.62. The VE of LNH on HIV-1 on the surface is the reduction of the titer by 2.3–3.3 lg TCID50, depending on disinfection duration, the HSV-1 reduction titer amounted to 3.8 lg TCID50.
Conclusion. For the first time it has been established that LNH and OLC possess a virucidal effect against both enveloped and non-enveloped viruses and can be considered as components of virucidal antiseptics. The VE of LNH without participation of additional compounds is very promising.
Full Text
Introduction
A pressing issue in modern medicine is the fight against infectious diseases, more than 80% of which are caused by viruses. There is currently no specific etiotropic antiviral therapy, nor are there vaccines for the prevention of many viral diseases. Therefore, protective preventive measures play a key role in the fight against viral infections, specifically the use of disinfectants with a virucidal effect (VE) — an action on the virus outside the cell.
Antiseptics with active ingredients such as ethyl (ethanol), isopropyl (propan-2-ol) and propyl (propan-1-ol) alcohols or their combinations in various ratios, as well as combinations of these alcohols with active ingredients from other groups of chemical compounds (MR 3.5.1/3.5.5.0355-24) [2]. The selection of active ingredients for antiseptics is complicated by limitations regarding the types of viruses sensitive to them and/or the duration of disinfection [3, 4]. Therefore, the search for new agents with pronounced VE and the shortest treatment duration appears to be a priority. Another challenge is selecting an appropriate model for testing the VE of antiseptics. For bacteria, the activity of substances is studied on the skin of volunteers’ hands, which is problematic in the case of viruses.
Formulations based on lanthanide rare earth metals have been found to possess both therapeutic and biocidal antimicrobial properties [5, 6]. These antiseptics have demonstrated bactericidal and bacteriostatic effects [7]. However, the available sources contain little information regarding their efficacy against human viruses [8]. The pharmacological and biological properties of lanthanum and lanthanide salts are being intensively studied, but many aspects of their effects on biological systems remain unclear.
The aim of the study is to investigate the VE of lanthanum nitrate hexahydrate (LNH) and compositions based on it to determine the possibility of their further use as components of topical antiseptics.
Materials and methods
Viruses
Human immunodeficiency virus type 1 (HIV-1), strain HIV-1899A, was obtained from the HIV strain collection of the D.I. Ivanovsky Institute of Virology at the N.F. Gamaleya National Research Center for Epidemiology and Microbiology. Virus titer — 7.0 lg 50% tissue culture infectious dose (TCID50).
Herpes simplex virus type 1 (HSV-1) was obtained from the State Virus Collection of the D.I. Ivanovsky Institute of Virology, N.F. Gamaleya National Research Center for Epidemiology and Microbiology. Virus titer — 5.0 lg TCID50.
Poliovirus, Sabin vaccine strain, type 1 (PV-1) was obtained from the M.P. Chumakov Research Institute of Poliomyelitis and Viral Encephalitis. Virus titer — 6.5 lg TCID50.
Human adenovirus, type 5 (AV-5) was obtained from the State Virus Collection of the D.I. Ivanovsky Institute of Virology at the N.F. Gamaleya National Research Center for Epidemiology and Microbiology. Virus titer — 6.5 lg TCID50.
Cells
The study utilized the MT-4 lymphoblastoid cell line for work with HIV-1, Vero green monkey kidney cell culture for work with PV-1 and HSV-1, and the passaged human cell line HEp-2 for work with AV-5. All cells were obtained from the collection of the D.I. Ivanovsky Institute of Virology at the N.F. Gamaleya National Research Center for Epidemiology and Microbiology.
Organic composition under study
We prepared an organic lanthanum composition (OLC) [11]: LNH — La(NO3)3·6H₂O (Lanhit Ltd.), triethylene glycol (TEG), ethyl carbitol (EC), glycerin, and NaOH. OLC and LNH were studied undiluted.
LNH with a salt concentration of 5.8 wt.% (133 mM/L) in water and glycols:
1,2-propylene glycol (PG) — C3H8O2, CH3-CH(OH)-CH2-OH;
1,3-butylene glycol (BG) — C4H10O2, CH3CH-OH-CH2CH2OH;
TEG — C6H14O4, HO-CH2-CH2-O-CH2-CH2-O-CH2-CH2-OH;
EC — C6H14O3, C2H5OCH2CH2OCH2CH2OH;
polyethylene glycol-400 (PEG) — C2nH4n+2On+1, НО–СН2–(СН2-СН2О)n–СН2–ОН.
Study structure
When using the in vitro suspension method, the test virus was mixed with the test samples in a 1:9 ratio; after incubation for 1–5 minutes, the mixture was added to a cell culture susceptible to the virus, a series of consecutive 10-fold dilutions was prepared, and the samples were incubated for 3–7 days (depending on the virus under study) in an incubator at 37°C in an atmosphere containing 5% CO2 and 98% humidity.
To determine the viral load of the test surfaces during decontamination using the “wiping” method, sterile test surfaces made of nitrile (synthetic rubber) (GOST R 52239-2004, sections 6.2, 6.3, 6.4; GOST 32337-2013) and eco-leather (cotton fabric, polyurethane) measuring 10 × 10 cm, onto which a viral suspension was applied at a rate of 0.5 mL with the addition of 5% inactivated bovine serum per 100 cm² and evenly distributed across the surface (MG 3.5. 2431-08, R 4.2.3676—20). The contaminated surfaces were air-dried until completely dry at a temperature of (20 ± 2°C) and relative humidity of 50–60%, after which they were treated with disinfectants.
Efficacy was assessed by collecting samples from treated surfaces 1–5 minutes after the virus came into contact with the surface. Bovine serum was used as the neutralizing agent. To determine the infectious titers of the virus, 96-well culture plates (Costar) were used. Each experimental point included 4 repetitions.
Recording of results
The cytopathic effect of the virus — morphological and functional degenerative changes in cell culture — was determined by light microscopy, comparing the changes with a control group of cells to identify cellular degeneration, destruction, and death caused by the virus replicating within them. The TCID50 was determined using the Reed-Mench total cumulative method. Statistical analysis of the results was performed using Student’s t-test.
Results
Treatment for 5 minutes in vitro reduced the infectious titer of PV-1 (4.50 ± 0.06 lg TCID50), AV-5 (4.00 ± 0.08 lg TCID50), HSV-1 (3.80 ± 0.07 lg TCID50), and HIV-1 (4.50 ± 0.05 lg TCID50).
In the PV-1 and AV-5 models, viral particles were detected on the test surfaces using the “wiping” method (Table 1).
Table 1. Results of the OLC VE study on PV-1 and AV-5 on test surfaces using the “wiping” method
Virus | Test surface | Processing time, min | Degree of viral inactivation, lg TCID50 |
PV-1 | Artificial skin | 1.0 | 4.50 ± 0.05 |
2.0 | 5.00 ± 0.07 | ||
Nitrile | 5.0 | 5.00 ± 0.06 | |
AV-5 | Artificial skin | 1.0 | 4.00 ± 0.06 |
2.0 | 4.50 ± 0.05 | ||
Nitrile | 5.0 | 4.50 ± 0.07 |
The use of LNH dissolved in H₂O, PG, TEG, and EC caused a decrease in the PV-1 titer by more than 4.0 lg TCID50. PG reduced the PV-1 titer by 4.0 lg ID50, while TEG and EC reduced it by 3.1–3.2 lg TCID50. With regard to HSV-1, all tested samples demonstrated a reduction in viral titer of 4.0–4.3 lg ID50 (Fig. 1).
Fig. 1. VE of LNH and glycol solutions against PV-1 and HSV-1 in an in vitro suspension assay.
1-minute exposure.
The results obtained indicate that LNH exhibited VE, reducing the HIV-1 titer by 5.30 ± 0.07 lg TCID₅₀ when dissolved:
- in PG — by 5,30 ± 0,07 lg TCID50;
- in BG — на 5,30 ± 0,06 lg TCID50;
- in PEG-400 — by 4,80 ± 0,04 lg TCID50;
- in EC — by 4,80 ± 0,05 lg TCID50.
OLC at pH ~ 7 exhibited VE, reducing the HIV-1 titer by 5.30 ± 0.07 lg TCID50 when combined with PG, by 5.30 ± 0.06 with BG, by 5.30 ± 0.05 with TEG, and by 4.30 ± 0.07 with PEG-400. The formulation with TEG and EC showed a reduction in viral titer by 3.50 ± 0.04 lg TCID50.
The OLC formulation with EC and TEG reduced the test virus titer by 4.8 lg TCID50 at pH < 4.5 (Fig. 2) and by 3.8 lg TCID50 at pH > 6.95.
Fig. 2. The relationship between the degree of HIV inhibition and changes in pH in a solution of an organic lanthanum-containing compound; in vitro suspension assay, 1-minute exposure.
The LNH based antiseptic exhibited VE against HIV-1 when used to treat surfaces contaminated with HIV, reducing the viral titer by 2.3–3.3 lg TCID50 depending on the duration of decontamination. In the case of HSV-1, the reduction was 3.8 lg TCID50, regardless of the duration of treatment (Table 2).
Table 2. Results of the H2O-based viral load assay for HIV-1 and HSV-1 on test surfaces using the “wiping” method
Virus | Test surface | Processing time, min | Degree of viral inactivation, lg TCID50 |
HIV | Artificial skin | 1.0 | 2.3 ± 0.04 |
2.0 | 3.3 ± 0.05 | ||
HSV | 1.0 | 3.8 ± 0.03 | |
2.0 | 3.8 ± 0.05 |
Discussion
Viruses constitute a distinct domain, Virae, comprising a vast number of families, genera, and classes, with tens of thousands of species, of which 600–800 cause human disease. Based on the type of genetic material, they can be divided into RNA- or DNA-containing viruses; based on the presence of a lipid envelope, into enveloped and non-enveloped viruses; by the number of nucleic acid strands — into single-stranded or double-stranded [1, 2].
Non-enveloped viruses were selected as test viruses for the studies: poliovirus (single-stranded RNA-containing) and adenovirus (double-stranded DNA-containing), which are essential for determining the VE of disinfectants and exhibit high resistance to various groups of chemicals used in disinfection [9, 10]. HIV-1 (single-stranded RNA-containing) and HSV-1 (double-stranded DNA-containing) were used as enveloped test viruses.
A number of viruses affect the skin, including HSV-1 and HSV-2, herpes zoster, papillomavirus, smallpox, chickenpox, and measles viruses, among others. Herpes infection is widespread, being one of the most common human infections. The main “entry points” for HSV are the skin and mucous membranes. The virus is epithelial-tropic and enters the cell via receptor-mediated endocytosis, where it replicates [1, 11].
HIV, which is classified as a socially significant pathogen, also enters the body through mucous membranes or broken skin. It is commonly believed that HIV is a “weak” virus that dies quickly outside the body. This is not entirely true: under certain conditions, it remains infectious in the environment for more than two weeks. Furthermore, it is highly resistant to ultraviolet radiation [2]. Therefore, for our study, we additionally used HIV-1 and HSV-1 as test viruses.
Virus inactivation is assessed by the loss of its infectious activity. A degree of viral inhibition exceeding 4.0 lg TCID50 provides grounds for considering disinfectants effective, according to current regulatory documentation. For testing skin disinfectants, standards recommend the use of the suspension method (in vitro), which we employed, and the method using batiste test surfaces. The latter, in our opinion, is not sufficiently informative; therefore, artificial skin was used as the test surface, which more adequately mimics the situation with human skin.
Lanthanides possess a range of biological effects, including immunomodulatory and antibacterial activity [12–17]. We found no information on the biological effects of lanthanides. In this study, we investigated the VE of LNH and compositions based on it.
During the study of OLC application in in vitro experiments, a significant reduction in the titers of the following viruses was observed: PV-1, AV-5, HSV-1, and HIV-1 — from 3.8 to 4.5 lg TCID50. The smaller reduction in HSV-1 correlates with its lower titer. In the PV-1 and AV-5 model on test surfaces — artificial skin and nitrile (medical gloves) — a reduction in the test virus titer of 4.0–5.0 lg TCID50 was observed. As exposure time on the test surfaces increased, the degree of reduction for both PV-1 and AV-5 increased.
The COVID-19 pandemic has triggered a new wave of research into glycols for use in antiseptic and antiviral agents, as well as for standalone applications. In particular, PG exhibits activity against the influenza virus in liquid and aerosol forms, as well as against SARS-CoV-2 [18, 19]. Since, in addition to LNH, OLC contains several components, including glycols, it was of interest to investigate the virucidal activity of these components individually [20].
The results obtained showed that, with regard to non-enveloped PV-1, which is more difficult to inactivate with chemical disinfectants, there is a clear difference in the virucidal activity of the active ingredients. PG reduced the PV-1 titer by 4.0 lg TCID50, while TEG and EC reduced it by 3.1–3.2 lg TCID50, which is not sufficiently effective; however, this represents a 103-fold reduction in infectivity. With regard to HSV-1, LNH and all glycols demonstrated a reduction in viral titer by 4.0–4.3 lg TCID50.
LNH not only exerts a pronounced VE against PV-1 but also enhances the action of the glycols through its presence in the formulation. Thus, the combined use of these ingredients demonstrates a synergistic VE.
In a study using an HIV-1 model, it was found that GNL dissolved in glycols exhibited VE, reducing the HIV-1 titer by 4.8–5.3 lg TCID50 at pH < 4.62. OCL with PG, BG, and PEG-400 instead of TEG and EC at pH ~7 exhibited VE, reducing the HIV-1 titer by 4.3–5.3 lg TCID50 within 1–2 minutes. At the same time, the OLC with EC and TEG showed a reduction of 3.5 lg TCID50.
It is known that pH can influence the VE and bactericidal activity of substances [9]. Therefore, it was of interest to study the VE of these compounds at different pH values using an HIV-1 model. It was found that OLC with EC and TEG at pH < 4.5 reduced the HIV-1 titer by 4.8 lg TCID50, and at pH > 6.95 — by 3.8 lg TCID50. In the acidic range, OLC proved to be more effective.
The preparation based on LNH in H₂O exhibited VE against HIV-1 and HSV-1 not only in in vitro experiments but also when treating surfaces contaminated with viruses, reducing their titers by 2.3–3.8 lg TCID50.
Conclusion
These preliminary findings suggest that both LNH and various compositions based on it exhibit VE against both enveloped and non-enveloped viruses.
Standards recommend a disinfection duration of no more than 2–3 minutes for topical antiseptics; therefore, the fact that the tested formulations achieve VE within 1–2 minutes justifies considering them as components for the development of virucidal disinfectants.
The detection of virucidal activity in LNH when used alone without additional substances (various glycols) opens up interesting prospects for the development of new antiseptics active against viral pathogens.
About the authors
Elena V. Berezhnaya
National Research Center of Epidemiology and Microbiology named after honorary academician N.F. Gamaleya
Author for correspondence.
Email: elvenel@gmail.com
ORCID iD: 0000-0002-1551-718X
junior researcher, Laboratory of antivirals and disinfectants, D.I. Ivanovsky Institute of virology
Russian Federation, MoscowIgor I. Pashkin
MIREA — Russian Technological University
Email: pashkin@mirea.ru
ORCID iD: 0000-0002-5315-6370
Cand. Sci. (Chem.), main specialist, S.S. Medvedev Department of chemistry and technology of macromolecular compounds, M.V. Lomonosov Institute of Fine Chemical Technologies
Russian Federation, MoscowMarina S. Bochkova
National Research Center of Epidemiology and Microbiology named after honorary academician N.F. Gamaleya
Email: bochkovams1@yandex.ru
ORCID iD: 0000-0001-9295-8379
leading researcher, Laboratory of antivirals and disinfectants, D.I. Ivanovsky Institute of Virology
Russian Federation, MoscowNina G. Kondrashina
National Research Center of Epidemiology and Microbiology named after honorary academician N.F. Gamaleya
Email: nina.kond1950@yandex.ru
ORCID iD: 0000-0003-3985-3839
leading researcher, Laboratory of antivirals and disinfectants, D.I. Ivanovsky Institute of Virology
Russian Federation, MoscowGalina M. Kuzmicheva
MIREA — Russian Technological University
Email: galina_kuzmicheva@list.ru
ORCID iD: 0000-0003-4458-8013
Dr. Sci. (Chem.), Professor, Department of materials engineering, Advanced Engineering School of Microwave Electronics, Director, Scientific and Educational Center “Multiscale Design of Materials”
Russian Federation, MoscowElena N. Domoroshchina
MIREA — Russian Technological University
Email: elena_domoroshchina@mail.ru
ORCID iD: 0000-0001-6082-7331
Cand. Sci. (Chem.), Head, Specialized educational and scientific laboratory “Functional materials and nanostructures”, Scientific and Educational Center “Multiscale Design of Materials”
Russian Federation, MoscowDmitry N. Nosik
National Research Center of Epidemiology and Microbiology named after honorary academician N.F. Gamaleya
Email: dnnosik@yandex.ru
ORCID iD: 0000-0001-5757-5671
Dr. Sci. (Med.), Professor, Head, Laboratory of antiviral and disinfection agents, D.I. Ivanovsky Institute of Virology
Russian Federation, MoscowReferences
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