INTRODUCTION:

Lomelosia isetensis (L.) Soják (lomelosia isetensis) belongs to the Caprifoliaceae Juss family. Lomelosia isetensis (L.) Soják is a low semi-shrub up to 45 cm in height. It grows in Central Kazakhstan, forming thickets suitable for harvesting raw materials. It is also found in western Kazakhstan and the southern Kazakhstan regions^1-2, the Caucasus, Central Asia and Siberia³. The medicinal properties of many herbal preparations used in folk medicine are due to the presence of organic and inorganic compounds with various types of biological activity.

The overall effect of the extract is not limited to the components that prevail in the main composition; traces of components in small quantities also affect the biological activity, inhibiting or, conversely, enhancing the effect of individual substances, which contributes to the synergistic effect. Studies of the antiradical activity of the carbon dioxide extract of Lomelosia isetensis (L.) Soják (1.8-cineol - 29.1%, α-santonin - 10.6%, α-thujone - 9.8%, β-thujone - 5.0%) have shown low antiradical activity compared with butylhydroxyanisole^4-6. The oil extract of Lomelosia isetensis (L.) Soják, obtained under ultrasound field conditions, showed high activity against test strains of Escherichia coli and the clinical strain of Candida albicans⁷. The toxicity of the CO₂-extract from the herb Lomelosia isetensis was studied on the basis survival rate of the euryhaline crustacean Artemia salina L. (Branchiopoda, Crustacea). When determining the level of toxicity (GOST 31959- 2012 (ISO 14669: 1999), it was established that the carbon dioxide extract from the herb Lomelosia isetensis (L.) Soják in a concentration of 10mg/ml (92, A, %) and 5mg/ml (85, A, %) - "has acute toxicity" – wich is highly toxic; at concentration of 1mg/ml - "does not have acute toxicity" - slightly toxic (19, A, %). In accordance with Figure 1, a diagram is presented based on the average toxicity indices of the carbon dioxide extract from the herb Lomelosia isetensis (L.) Soják.

Figure 1. Acute toxicity of CO₂-extract from the herb Lomelosia isetensis (L.) Soják. at different concentrations

The mineral composition of the aboveground parts of the lomelosia isetensis plants was studied via atomic emission spectrometry with inductively coupled plasma, and the results are presented in Table 1. Lomelosia isetensis (L.) Soják contains 59 minerals, of which 9 are macronutrients, 31 are trace elements and 19 are ultramicroelements. The quality indicator, total ash, reflects the sum of the mineral substances in Lomelosia isetensis (L.) Sojak. was determined to be within 9.02%. The total ash content was determined according to the method described in the State Pharmacopoeia of the Republic of Kazakhstan, Vol. 1, p. 129. Numerical indicators of the content of radionuclides (Cs-137 Bq/kg, Sr-90 Bq/kg) in the herb of Lomelosia isetensis are presented in Table 2. The radiation safety indicators comply with the requirements of the regulatory documentation (RD) "On approval of hygienic standards for ensuring radiation safety" Order of the Minister of Health of the Republic of Kazakhstan dated August 2, 2022 No. ҚР ДСМ-71. The main component of the CO₂-extract of the herb Lomelosia isetensis (L.) Sojak. are as follows: 1,8-cineol -29.1%; α-santonin - 10.6%; α-thujone - 9.8%; β-thujone - 5.0% and campesterol - 1,8%⁸. The components α-santonin, α-thujone, and β-thujone are toxic⁹, but santonin was excluded from the list of biologically active substances with undesirable toxicological effects¹⁰. The content of thujone is regulated by the Decision of the Council of the Eurasian Economic Commission of July 20, 2012 No. 58, On the adoption of the technical regulations of the Customs Union "Safety requirements for food additives, flavorings and technological aids", On amendments to the technical regulations of the Customs Union "On the safety of perfumery and cosmetic products" (TR CU 009/2011) Decision of the Council of the Eurasian Economic Commission of March 29, 2019 No. 32.

Table 1. Mineral composition of ash from the herb Lomelosia isetensis (L.) Soják

No.	Element	Symbol	Content (mg/kg)	No.	Element	Symbol	Content (mg/kg)
1	2	3	4	1	2	3	4
1	Lithium	Li	6,842	30	Cadmium	Cd	<0,01
2	Beryllium	Be	0,121	31	Indium	In	0,003218
3	Boron	B	32,23	32	Tin	Sn	1,933
4	Sodium	Na	2276	33	Antimony	Sb	0,02922
5	Magnesium	Mg	7673	34	Cesium	Cs	0,02418
6	Aluminum	Al	827,5	35	Barium	Ba	228,7
7	Phosphorus	P	3122	36	Lanthanum	La	0,265
8	Potassium	K	31820	37	Cerium	Ce	0,6546
9	Calcium	Ca	62780	38	Praseodymium	Pr	0,2686
10	Scandium	Sc	0,7488	39	Neodymium	Nd	1,048
11	Vanadium	V	1,068	40	Samarium	Sm	0,3424
12	Chromium	Cr	24,9	41	Europium	Eu	0,64
13	Manganese	Mn	73,95	42	Halinium	Gd	0,1435
14	Iron	Fe	1054	43	Terbium	Tb	0,03217
15	Cobalt	Co	0,53	44	Dysprosium	Dy	0,04437
16	Nickel	Ni	5,867	45	Holmium	Ho	0,01496
17	Copper	Cu	25,85	46	Erbium	Er	0,04339
18	Zinc	Zn	17,07	47	Thulium	Tm	0,0198
19	Galium	Ga	1,244	48	Ytterbium	Yb	0,04742
20	Germanium	Ge	0,05013	49	Lutetium	Lu	<0,01
21	Arsenic	As	0,58	50	Hafnium	Hf	0,07482
22	Selenium	Se	0,7238	51	Tantalum	Ta	0,02255
23	Rubidium	Rb	11	52	Tungsten	W	0,2649
24	Strontium	Sr	225	53	Rhenium	Re	0,003731
25	Yttrium	Y	1,398	54	Mercury	Hg	0,003053
26	Zirconium	Zr	5,513	55	Titanium	Ti	0,008772
27	Niobium	Nb	0,3317	56	Lead	Pb	1,33
28	Molybdenum	Mo	1,12	57	Bismuth	Bi	0,00953
29	Silver	Ag	0,7274	58	Thorium	Th	0,03181
				59	Uranium	U	0,1394

Table 2. Radiation safety index for Lomelosia isetensis (L.) Soják.

Name of indicator, unit of measure	Lomelosia isetensis (L.) Soják.	Norms according to the regulatory document
Cs-137 Bq/kg	˂7	˂400 Bq/kg
Sr-90 Bq/kg	˂13	˂200 Bq/kg

The content of heavy metals in the aboveground part of Lomelosia isetensis (L.) Soják does not exceed the permissible standards.

In accordance with Table 3, the transition of heavy metals in the chain of plant material – the CO₂ extract of Lomelosia isetensis (L.) Soják is presented. The content of heavy metals does not exceed the standard quality indicator (according to the State Pharmacopoeia of the Republic of Kazakhstan, Vol. 3).

The disk diffusion method is based on the inhibition of the surface, and visible growth of strains on a dense nutrient medium by an antimicrobial substance. The concentration differences of the test substance in a dense nutrient medium (agar) are achieved as a result of diffusion from an introduced paper disk (carrier). This carrier, impregnated with the test substance, was placed directly on the surface of the nutrient medium immediately after inoculation of the culture, via the "lawn" method.

Stages of the study:

- Preparation and composition of the nutrient medium;

- Preparation of a suspension of the test microorganism at a given concentration;

- Minimizing of the time interval between inoculation of the microorganism and application of disks to the surface of the medium;

- Compliance with the temperature and time regime of incubation¹¹.

MATERIALS AND METHODS:

The work on the study of the antimicrobial activity of CO₂-extract in relation to gram-positive strains, Staphylococcus aureus (АТСС 6538) and Bacillus subtilis (АТСС 6633); gram-negative strains of Escherichia coli (АТСС 25922) by the diffusion method on agar disks impregnated with a solution of the substance under study (GPhA.1.2.4.0010.15 Determination of the antimicrobial activity of antibiotics by the diffusion method in agar)^12-21 was carried out on the basis of the educational microbiological laboratory of the Department of Biomedicine of the "KMU" NCJSC.

The object of the study was carbon dioxide extracts obtained from the herb Lomelosia isetensis (L.) Soják²².

Comparison preparations:

Benzylpenicillin sodium. C₁₆H₁₇N₂Na0₄S. (Mr 356,4). 1011000 [6957-8]. (SPh RK vol. 2, p. 133). Indicator disks with benzylpenicillin, 10 U, 100 pcs. TC 9398-001-39484474-2000. RU No. FSR 2009/06472. Research Center of Pharmacopoeia, St. Petersburg.

Distilled water. 1095504. [7732-18-5]. (SPh RK vol.1, p. 347).

Solvents:

“Medium Eagle fluidum steril”, “Eagle medium”, without glutamine, for cell culture (500±25) ml. Series No. 735.

Ethanol (96%) was used as a control. C₂H₆O. (Mr 46.07). 102500. [64-17-5]. (SPh RK vol. 2, p. 581).

Microorganisms:

gram-positive strains of Staphylococcus aureus, Bacillus subtilis, and a gram-negative strain of Escherichia coli from the Republican State Enterprise on the Right of Economic Management "Republican Collection of Microorganisms" of the Science Committee of the Ministry of Education and Science of the Republic of Kazakhstan.

Materials:

Chistovich agar (yolk-salt agar); Endo agar; nutrient agar; alcohol burner; test tubes; turbidity standard for 5 units; TV-80-1 dry-air thermostat, 0.5 cm diameter filter paper disks, bacteriological loops, tweezers, 100–1000 μl digital pipette.

Statistical processing of the results of the biological activity studies was carried out via parametric statistical methods with calculation of the arithmetic mean and standard error.

Experimental:

1. A study of the antimicrobial activity of CO₂-extract from the herb Lomelosia isetensis (L.) Sojak in relation to gram-positive strains, Staphylococcus aureus and Bacillus subtilis; gram-negative strains of Escherichia coli.

Preparation of material for inoculation and seeding:

a) Carbon dioxide extract was dissolved in 96% ethyl alcohol (0.1 g per 3 ml of solvent);

b) The test tube was washed in "medium Eagle^,s fluidum sterile" medium and then an alcohol solution of CO₂- extract was added.

c) An 18-20 hour agar or 5-6 hour broth culture of the studied microorganism was used. The suspension was prepared from a sterile solution of isotonic sodium chloride, in which the culture was diluted to a turbidity standard of 0.5 McFarland, and then brought to a final concentration of 1-2х10⁷ CFU/ml (diluted 10 more times with isotonic solution);

d) The prepared inoculum was added to a Petri dish with 1-2 ml of nutrient medium, which was evenly distributed with a sterile spatula or shaken. Excess liquid was removed with a pipette. The slightly open dishes were dried at room temperature for 10-15 minutes. The disks were applied and placed at a distance of at least 15 mm from the edge of the dish and at least 30 mm between the disks. The mixture was incubated for 18-24 hours at 37°C. The samples were tested in three parallel experiments. The crops were incubated in a dry-air thermostat at 36.5–37.4 ^oC, and the growing cultures were counted after 18–24 hours.

RESULTS AND DISCUSSION:

A study of the antimicrobial activity of CO₂-extract from the herb Lomelosia isetensis (L.) Soják in relation to gram-positive strains, Staphylococcus aureus and Bacillus subtilis; gram-negative strains of Escherichia coli.

Antimicrobial activity was determined by the diameter of the growth inhibition zones (d) of test strains (mm). At d≤10 mm continuous growth in the dish it was assessed as the absence of antibacterial activity, 10-15 – weak activity, 15-20 mm – moderate activity, and at d>20 – pronounced activity. The growth inhibition diameters of the test strains are presented in Table 4 and Figure 1, where the test substance: CO₂-extract from the herb Lomelosia isetensis (L.) Soják is listed as 2.

Table 4. Diameters of growth inhibition of test strains

No.	Substances under study	*Bacillus subtilis* АТСС 6633	*Staphylococcus aureus* АТСС 6538	*Escherichia coli* АТСС 25922
1	CO₂-extract from the herb Lomelosia isetensis (L.) Soják (2)	14±1,0	18±1,0	13±1,0
2	Ethanol (96%)	11±1,0	10±1,0	10±1,0
3	Benzylpenicillin sodium	15±1,0	17±1,0	12±1,0

А B

Figure 1. (A) Nutrient agar for Bacillus subtilis, (B) Chistovich agar for Staphylococcus aureus; (C) Endo agar for Escherichia coli

CONCLUSION:

The results of studies on toxicity and antimicrobial activity reveled the prerequisites for a thorough comprehensive study of the safety and efficacy of medicinal products from Lomelosia isetensis (L.) Soják.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

This research has been/was/is funded by the Committee of Science of the Ministry of Science and Higher Education of the Republic of Kazakhstan (Grant No. AP19679527 and BR 24992761).

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Received on 15.12.2024 Revised on 14.06.2025

Accepted on 29.10.2025 Published on 13.01.2026

Available online from January 17, 2026

Research J. Pharmacy and Technology. 2026;19(1):333-337.

DOI: 10.52711/0974-360X.2026.00048

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Heavy metals	Lomelosia isetensis (L.) Sojak		The norms of the quality index mg/kg
Heavy metals			Vegetable raw materials and substances for pharmaceutical use
Cadmium (Cd)	<0,01	0,000221 ± 0,000066	1,0
Plumbum (Pb)	1,33	0,000375 ± 0,000112	5,0
Hydrargyrum (Hg)	0,0031	-	0,1
Arsenicum (As)	0,58	-	1,0
Zincum (Zn)	17,07	0,092990 ± 0,027900
Cuprum (Cu)	25,85	0,082130 ± 0,024640