АМИНОКИСЛОТНЫЙ СОСТАВ И КОЛИЧЕСТВО КОМПОНЕНТОВ КОРНЯ РАСТЕНИЯ Ferula lipskyi Korovin, ВЫРАЩЕННОГО В ФЕРГАНСКОЙ ДОЛИНЕ

УДК 547.9:581.192

Abstract

This article examines the amino acid composition and quantitative parameters of the roots of Ferula lipskyi, a plant native to the Fergana Valley. The relevance of this study lies in the richness of plants of the genus Ferula in biologically active substances, including amino acids, flavonoids, coumarins, and essential oils. Since these plants are widely used in folk medicine and the pharmaceutical industry, an in-depth study of their chemical composition is of significant scientific and practical importance. Ferula lipskyi roots, collected in May 2025 in the Fergana region, were used as the study object. Qualitative and quantitative amino acid analysis was performed by high-performance liquid chromatography using an Agilent 1260 II Infinity system and a fluorescence detector. Sigma Aldrich amino acid standards were used for the analysis. According to the study results, the amino acids serine, glycine, threonine, arginine, alanine, lysine, valine, methionine, phenylalanine, isoleucine, and proline were detected in the sample. The highest contents were observed for threonine (0.98 mg/g), glycine (0.79 mg/g), and lysine (0.53 mg/g). However, the amino acids asparagine, glutamine, histidine, glutamate, and tryptophan were not detected. The obtained results showed that the roots of Ferula lipskyi are rich in biologically and pharmacologically important natural compounds. In particular, the presence of essential amino acids such as threonine and lysine increases the biological value of the plant. The results of the study expand the possibilities of using this plant as a source of natural biologically active substances in the pharmaceutical, food, and biotechnology industries.

Аннотация

В данной статье изучается аминокислотный состав и количественные параметры корневых частей Ferula lipskyi, произрастающей в Ферганской долине. Актуальность исследования объясняется богатством растений рода Ferula биологически активными веществами, включая аминокислоты, флавоноиды, кумарины и эфирные масла. Поскольку эти растения широко используются в народной медицине и фармацевтической промышленности, углубленное изучение их химического состава имеет важное научное и практическое значение. В качестве объекта исследования была выбрана корневая часть растения Ferula lipskyi, собранная в мае 2025 года в Ферганской области. Качественный и количественный анализ аминокислот проводился методом высокоэффективной жидкостной хроматографии с использованием системы Agilent 1260 II Infinity и флуоресцентного детектора. В ходе анализа использовались стандартные образцы аминокислот от Sigma Aldrich. Согласно результатам исследования, в образце были обнаружены аминокислоты серин, глицин, треонин, аргинин, аланин, лизин, валин, метионин, фенилаланин, изолейцин и пролин. Наибольшее содержание наблюдалось у треонина (0,98 мг/г), глицина (0,79 мг/г) и лизина (0,53 мг/г). При этом аминокислоты аспарагин, глутамин, гистидин, глутамат и триптофан обнаружены не были. Полученные результаты показали, что корневые части Ferula lipskyi богаты биологически и фармакологически важными природными соединениями. В частности, наличие незаменимых аминокислот, таких как треонин и лизин, повышает биологическую ценность растения. Результаты исследования расширяют возможности использования этого растения в качестве источника природных биологически активных веществ в фармацевтической, пищевой и биотехнологической промышленности.

Keywords: Ferula lipskyi, genus Ferula, amino acid composition, plant roots, biologically active substances, high-performance liquid chromatography, fluorescence detector, glycine, lysine.

Ключевые слова: Ferula lipskyi, род Ferula, аминокислотный состав, корни растений, биологически активные вещества, высокоэффективная жидкостная хроматография, флуоресцентный детектор, глицин, лизин.

Introduction

 With the development of medicine, the delivery of medications to patients is becoming increasingly important. The majority of drugs used in medical practice are biologically active substances isolated from plant components. Plants of the genus Ferula are a valuable source of natural biologically active substances and are widespread in the arid and mountainous regions of Central Asia. Representatives of this genus contain essential oils, resins, coumarins, flavonoids, terpenoids, amino acids, and minerals [1,2]. Due to their rich chemical composition, Ferula plants exhibit anti-inflammatory, antimicrobial, antioxidant, antispasmodic, and restorative properties. In folk and traditional medicine, they are used to treat various diseases and are also considered promising raw materials for the pharmaceutical, food, and agricultural industries [3,4]. The chemical composition of Ferula plants depends on the species, growing conditions, climate, soil composition, and stage of plant development, necessitating their comprehensive phytochemical and pharmacological studies. In recent years, there has been a growing interest in studying the chemical composition and biological activity of Ferula species [5,6].

Objective of the work: The purpose of the work is to determine the amino acid composition and quantitative indicators of the root parts of the Ferula lipskyi plant growing in the Fergana Valley using high-performance liquid chromatography, to assess the presence of biologically active amino acids, and to scientifically substantiate the pharmaceutical and biotechnological significance of the plant.

About the plant

Ferula lipskyi is a large perennial plant, typically reaching a height of 1.5–2.5 meters. It is characterized by a strong putrid odor. Rhizome: thick, large, often coarse. It is highly branched underground. It secretes a yellowish milky sap (rich in substances with anti-inflammatory properties). Stem: erect, large, hollow, smooth or slightly inflamed. It reappears annually. Leaves: large, intricately divided into four or five parts. The lobes are small and needle-like. The leaf blades have an expanded, cap-shaped sheath. Flowers: small, yellowish. Arranged in complex umbel-shaped (zonal) inflorescences. The flowers are bisexual. Fruit: A mericarp (split fruit) dividing into two wing-shaped parts. Each part has five distinct marginal lines. The fruit turns yellow-orange when ripe. Distribution and ecology: Distribution: Central Asia, especially the foothills and mountains of Uzbekistan, Tajikistan, and Kazakhstan. Habitat: Dry valleys, rocky and sandy soils, steppes, and slopes. Growing season: Grows in spring; fruits ripen in late summer. The root of this plant is characterized by its richness in biologically active substances, especially amino acids [7,8]. Researchers have noted that the chemical composition of Ferula species depends on climate, soil, and vegetation stage [2,9].

Materials and methods

The root parts of Ferula lipskyi, collected in the Fergana district of the Fergana region in May 2025, were used as the test sample. The plant sample was analyzed for qualitative and quantitative amino acid parameters using standard samples supplied by Sigma Aldrich (Germany). The determination was carried out using a fluorescence detector (FLD) installed on an Agilent 1260 II Infinity liquid chromatography system, manufactured by Agilent Technologies, USA [10]. A Poroshell 120 EC-C18 column (150 mm × 4.6 mm × 4 μm), manufactured in the USA, was used as the stationary phase. Pre-column derivatization was performed in an automatic programmable mode. For amino acid analysis, the following mobile phases were used: phase A — a solution of sodium dihydrogen phosphate (40 mM), pH 7.8, and phase B — a mixture of acetonitrile, methanol, and water (in a ratio of 45:45:10), using a gradient mode. HPLC is one of the most effective analytical methods for the determination of amino acids in medicinal plants [11].Time | Phase A % (sodium dihydrogen phosphate solution, 40 mM, pH 7.8) | Phase B% (acetonitrile: methanol: water = 45:45:10). The flow rate was 1 ml/min, the thermostat temperature was 40°C, the injected sample volume was 5 μl, the analysis was carried out for 30 minutes, and the following chromatograms were obtained.

Figure 1. Chromatogram obtained from standard samples of 16 different amino acids

Hydrolysis of the amino acids contained in the sample was carried out as follows:First, all samples were ground to a size of 1 mm. A 5.0 g sample was ground and weighed to the nearest 0.001 mg using an analytical balance (FA220 4N). Next, the sample was placed in a 200 ml flask with a reflux condenser, 50 ml of hydrochloric acid (6 N) was added, and the flask was placed in a thermostat at 110°C. Hydrolysis was carried out for 24 hours with magnetic stirring. After hydrolysis was complete, the solution was cooled to room temperature, 10 ml was removed, and centrifuged for 10 minutes at 12,000 rpm. Then, 5 ml of the centrifugate was removed and neutralized with 6 N sodium hydroxide.After this, 1 ml of the solution was filtered through a filter with a pore size of 0.45 μm, placed in a vial and the following results were obtained.

Results and Discussion

The results obtained are presented in Table 1.

Table 1. Composition and quantity of amino acids of the root parts of the plant Ferula lipskyi, growing in the Fergana Valley

Figure 2. Chromatograms of amino acids obtained from samples of the root part of the plant Ferula lipskyi

The results of the study revealed that the amino acids serine, glycine, threonine, arginine, alanine, lysine, valine, methionine, phenylalanine, isoleucine and proline were present in the sample. The highest levels were observed in threonine, glycine and lysine. These results are consistent with studies conducted on other Ferula species [1,5,12]. The presence of essential amino acids such as threonine and lysine increases the biological value of the plant [13]. The high content of glycine indicates that it plays an important role in the synthesis of biologically active compounds [14].

Conclusion

Thus, the study of the amino acid composition of the roots of Ferula lipskyi, a plant grown in the Fergana Valley, revealed the presence of a number of physiologically important amino acids. High-performance liquid chromatography (HPLC) identified and quantified serine, glycine, threonine, arginine, alanine, lysine, valine, methionine, phenylalanine, isoleucine, and proline in the sample. However, asparagine, glutamine, histidine, glutamate, and tryptophan were not detected in the sample. Among the identified amino acids, the highest concentrations were found for threonine (0.98 mg/g), glycine (0.79 mg/g), and lysine (0.53 mg/g). The relatively high content of these amino acids indicates the biochemical value of Ferula lipskyi roots. Threonine is an essential amino acid and plays a key role in protein metabolism, immune system function, and maintaining metabolic processes in the body. Glycine is involved in the synthesis of biologically active compounds, and lysine is an important component of protein metabolism and tissue growth. A comparative analysis with representatives of the genus Ferula shows that the amino acid profile of Ferula lipskyi has its own distinctive features. Like other species of this genus, the plant contains a complex of biologically active substances, but the quantitative distribution of amino acids depends on the species, environmental conditions, soil composition, climatic factors, vegetation phase, and the plant part used. The predominance of threonine, glycine, and lysine in the roots of Ferula lipskyi distinguishes this species from several other representatives of the genus Ferula, for which variable accumulation of amino acids and other nitrogen-containing compounds is more often noted in the literature. The obtained results confirm that Ferula lipskyi is a promising plant material for further phytochemical and pharmacological study. The presence of essential and nonessential amino acids in the root parts of the plant allows this species to be considered a potential source of natural biologically active compounds that can be used in the pharmaceutical, food, and biotechnology industries. However, the absence of asparagine, glutamine, histidine, glutamate, and tryptophan in the studied sample indicates the specificity of the amino acid composition of Ferula lipskyi and the need for further research using samples collected at different stages of the growing season and from different growing regions. This will allow for a more complete assessment of the chemical composition of the plant, the establishment of patterns of amino acid accumulation, and the determination of its practical significance among other species of the genus Ferula.

The results confirmed that the root parts of Ferula lipskyi are rich in biologically active amino acids. This plant can be considered as a promising natural resource for the pharmaceutical, food and biotechnology industries [6,15]. However, further research on the amino acid composition of the plant in terms of growing season and region is desirable [2,9].

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