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DOI - 10.32743/UniTech.2025.130.1.19164
DEVELOPMENT OF THE SYNTHESIS PROCESS FOR ELLAGIC ACID EXTRACTION FROM MACLURA POMIFERA AND ITS CHROMATOGRAPHIC ANALYSIS
Sarvinoz Nazarova
PhD student
at the Tashkent institute of chemical technology, Uzbekistan, Tashkent E-mail: [email protected]
Xumora Abdashimova
Master's student
at the Tashkent institute of chemical technology Technology,
Uzbekistan, Tashkent E-mail: [email protected]
Golib Nazarov
Associate Professor (PhD) at the Tashkent institute of chemical technology Technology,
Uzbekistan, Tashkent E-mail: [email protected]
Anvar Normatov
Professor (PhD)
at the Tashkent institute of chemical technology Technology,
Uzbekistan, Tashkent E-mail: [email protected]
РАЗРАБОТКА ПРОЦЕССА СИНТЕЗА ДЛЯ ИЗВЛЕЧЕНИЯ ЭЛЛАГОВОЙ КИСЛОТЫ ИЗ МАКЛЮРЫ ПОМИФЕРЫ И ЕЕ ХРОМАТОГРАФИЧЕСКИЙ АНАЛИЗ
Назарова Сарвиноз Ботир кызы
аспирант
Ташкентского химико-технологического института,
Узбекистан, г. Ташкент
Абдашимова Хумора Аброровна
магистрант
Ташкентского химико-технологического института,
Узбекистан, г. Ташкент
Назаров Голиб Абдишукур оглы
канд. физ. - мат. наук, доцент Ташкентского химико-технологического института,
Узбекистан, г. Ташкент
Норматов Анвар Мирзаевич
профессор
Ташкентского химико-технологического института,
Узбекистан, г. Ташкент
ABSTRACT
This scientific article discusses the research and results of the extraction and chromatographic analysis of ellagic acid from the fruit of Maclura pomifera. The study utilized an acidic synthesis method to extract pure ellagic acid from Maclura pomifera for the first time. This method was found to be more efficient in terms of compound yield compared to previously used extraction methods, as demonstrated by quantitative analysis. The quality and quantity of the extracted ellagic acid were analyzed using Thin Layer Chromatography (TLC) and High-Performance Liquid Chromatography (HPLC). It was determined that the pure ellagic acid content in the extracted mixture was 88.18%.
Библиографическое описание: DEVELOPMENT OF THE SYNTHESIS PROCESS FOR ELLAGIC ACID EXTRACTION FROM MACLURA POMIFERA AND ITS CHROMATOGRAPHIC ANALYSIS // Universum: технические науки : электрон. научн. журн. Nazarova S.B. [и др.]. 2025. 1(130). URL: https://7universum.com/ru/tech/archive/item/19164
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UNIVERSUM:
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АННОТАЦИЯ
В данной научной статье освещены исследования по выделению эллаговой кислоты из плодов Maclura pomífera и ее хроматографическому анализу и их результаты, в том числе выделение чистой эллаговой кислоты из растения Maclura pomífera впервые методом кислотного синтеза в Выделение эллаговой кислоты. Количественные анализы показали, что этот метод отличается от используемых методов разделения эллаговой кислоты некоторой эффективностью при получении вещества. Качественный и количественный анализ выделенной эллаговой кислоты изучали методами тонкослойной хроматографии (ТСХ) и высокоэффективной жидкостной хроматографии (ВЭЖХ) и установили, что содержание эллаговой кислоты в чистой смеси составило 88,18%.
Keywords: Maclura pomifera, ellagic acid, chromatography, extract, gallic acid, acidic synthesis, bioactive compounds.
Ключевые слова: Maclura pomifera, эллаговая кислота, хроматография, экстракт, галловая кислота, кислотный синтез, биологически активные вещества.
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Introduction. One of the major challenges facing the global community today is finding solutions to environmental problems and addressing them effectively. Due to the continuous growth of the population, advancements in science and technology, and the increasing demands of human needs, numerous environmental issues are arising on Earth. According to data from the World Health Organization, the deterioration of environmental conditions has led to an increase in health issues, with more people suffering from various diseases in recent years.
To address these environmental challenges, protect the environment, and improve public health, Uzbekistan has designated the year 2025 as the Year of Environmental protection and green economy. This initiative highlights the importance of these issues and demonstrates the ongoing efforts to tackle them. The study of medicinal plants found in different regions of the country, their cultivation, and the extraction of various preparations from them can contribute significantly to solving these problems.
Review of Literature
Preventing and treating various health threats, as well as developing and applying therapeutic agents, is one of the key issues in contemporary science. Particularly, the development of biologically active supplements based on natural resources and evaluating their effects on living organisms is recognized as one of the main goals of modern scientific research.
Plants are rich in many biologically active compounds that are beneficial for health. The use of phyto-preparations in both global scientific research and medical practice has been rapidly expanding. In this regard, investigating the new therapeutic properties of plants, analyzing those plant sources whose biologically active compounds have not yet been studied, and developing optimal conditions and methods for extracting various bioactive substances from them are of great importance.
Today, natural plants have become one of the most active elements of traditional medicine. According to statistical data, a large portion of the world's population uses plants with medicinal properties. At present, the variety of medicinal plants is increasing, and traditional medicine has become enriched with medicinal
plants. One such plant, commonly cultivated as an ornamental tree in the streets and boulevards of our country, is Maclura pomifera.
Maclura pomifera (Moraceae) is a perennial tree species belonging to the mulberry family. It is a distinctive fruit-bearing tree that is found in various regions of our country and has been recognized as a part of the country's plant decoration for many years. The tree typically grows to a height of 8-15 meters. The fruits ripen in October-November and are spherical in shape, with an average diameter of 15-30 cm and a weight ranging from 300 to 600 grams [1].
Initial studies and global research have proven that Maclura pomifera fruits contain a significant amount of proteins, carbohydrates, lipids, amino acids, vitamins, flavonoids, triterpenoids, steroids, and other biologically active substances essential for the organism.
Biologically active supplements derived from Maclura pomifera fruits have demonstrated high antioxidant activity. Additionally, they exhibit antibacterial, antifungal, antiviral, and anti-inflammatory properties [2].
Maclura pomifera also contains a variety of other bioactive compounds necessary for living organisms, such as coloring agents, flavoring substances, and 25 types of macro and microelements, including potassium, calcium, sodium, magnesium, iron, chromium, boron, along with specific amounts of other chemical substances.
The gall acids and their residues found in Maclura pomifera fruits are considered beneficial for detoxification, cardiovascular diseases, skin cancer prevention, improving bone health, treating intestinal diseases, and addressing dental and gum issues. They are also useful for alleviating wrinkles and other signs of aging, sore throat, and cough, as well as acting as natural moisturizers and sun protection agents [3].
Ellagic acid content in the fruit of Maclura pomifera is relatively higher compared to other plants due to the presence of gall acid residues. Ellagic acid is a yellow-brown crystalline substance with the molecular formula C14H6O8. It is poorly soluble in water but dissolves well in organic solvents such as ethanol and acetone. Its melting point ranges from 360-380°C, and its density is 1.75 g/cm3.
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Due to its biological activity, ellagic acid is used as a treatment for various diseases. One of its healing properties is its antioxidant activity, which helps prevent oxidative damage to cells and reduces inflammation. Research has shown that ellagic acid helps prevent the growth of cancer cells and reduces the risk of certain types of cancer, such as breast, colon, and prostate cancer [4].
Ellagic acid is used in various fields, such as pharmaceuticals, cosmetics, and food supplements. It is commonly included in antioxidant supplements to combat damage from free radicals in the body. In cosmetics, ellagic acid is used for its anti-aging properties as it helps prevent the breakdown of collagen in the skin. In food, it is used as a natural preservative due to its antioxidant properties and can be found in products such as jams, jellies, and fruit juices [5].
Aim of the study
The aim of this research is to extract ellagic acid from Maclura pomifera fruit using an acidic synthesis method and conduct physical-chemical analyses of the obtained samples.
Research methods
Extraction of ellagic acid from Maclura pomifera fruit.The fruits of Maclura pomifera are first chopped into small pieces and dried until the moisture content reaches 8-10%. The dried sample is then ground into 1020 mm pieces. A 100g sample of the ground fruit
is mixed with 1000 ml of 20% ethyl alcohol and subjected to extraction at 80°C for 2 hours with continuous stirring (DLAB OS20-S) at 400 rpm. The resulting sample is centrifuged for 15 minutes at 5000 rpm, and the sediment is separated. The liquid portion is then evaporated in a rotary evaporator (65 rpm and 37°C) until 2/3 of the liquid is evaporated.
To the remaining liquid, a 5% sulfuric acid solution is added in a 1:5 ratio, and a concentration reaction environment is created at 105°C, initiating acidic hydrolysis. This process is carried out in a closed system using a round-bottom flask equipped with a condenser and heated by a water or sand bath. The total duration of the acid synthesis process is 5 hours. The resulting mixture of sediment and liquid is then fractionated using filter paper in a Büchner funnel.
The remaining thick portion on the surface is dried and then washed with methanol in the next stage to purify it. The purification process is repeated twice until the product reaches its pure form. The obtained ellagic acid is then dried at 60°C in a thermostat.
Ellagic acid composition analysis
The composition of the obtained ellagic acid was initially analyzed using Thin Layer Chromatography (TLC), and both Thin Layer Chromatography and HighPerformance Liquid Chromatography (HPLC) methods were widely used for further analysis. The principles of these methods are explained below.
Table 1.
Optimal composition of systems for analyzing ellagic acid using thin layer chromatography
№ Chemical Reagents for Thin Layer Chromatography System Amount of Substances (ml)
1 Acetonitrile 2
2 Acetone 2
3 Phosphoric acid 1
4 Toluene 3
5 Distilled water 0.5
The analysis process using chromatography was carried out with a SIGMA-ALDERICH plate. Initially, a system was prepared using precise amounts of the chemical reagents listed in Table 1, and the extract was applied to the plate in 1 cm intervals. The plate was then placed in a column for 10 minutes. After removal from the column, the plate was dried for 10 minutes and sprayed with anisaldehyde. Under the effect of anisaldehyde, a pink-colored compound appeared at the finish line of the plate, two yellow-green compounds appeared in the middle, and dark green-colored compounds appeared at the start line, as shown in Figure 2.
The high-performance liquid chromatography (HPLC) method was used to analyze ellagic acid qualitatively. For the identification of steroids and flavonoids via HPLC, phosphate and acetate buffer systems, as well as acetonitrile, were used as eluents. In our study, we employed a phosphate buffer system and acetonitrile to analyze the pure isolated ellagic acid.
Chromatographic Conditions:
Chromatograph: Agilent-1200 (equipped with an auto-injector)
Column: Exlipse XDB C18 (reverse-phase), 5 ^m, 4.6 x 250 mm
Detector: Diode array detector (DAD), identification at 254 nm and 276 nm Flow rate: 0.8 ml/min Eluent: Phosphate buffer : Acetonitrile 0-5 min: 95:5 6-12 min: 70:30
12-13 min: 50:50
13-15 min: 95:5
Thermostat temperature: 30°C, injected amount: 10 ^L
Results and Analysis: The raw materials and technological processes used for extracting ellagic acid are significant due to their high yield and low number of steps.
As a result, ellagic acid stands out from other compounds in this group due to its low cost and ease of use.
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AHBapb, 2025 r.
After the synthesis process is complete, the compound is purified twice using methanol and then dried. The obtained product appears brown in color. Figure 1 below shows three different samples of ellagic
acid, illustrating the differences between unpurified and purified ellagic acid.between unpurified and acid.
1
2
3
Figure 1. Samples of Ellagic Acid: 1. Initial sample after synthesis (1.5 g); 2. Sample after the first purification step (1.2 g); 3. Sample after the second
purification step (1 g)
In each purification step of ellagic acid, a noticeable decrease in the amount of the compound was observed,
with the final purified product amounting to 1 gram. Further studies were conducted to carry out both qualitative and quantitative analyses of the isolated ellagic acid.
Figure 2. HPLC Chromatogram of ellagic acid
Under the effect of anisaldehyde, a pink-colored compound appeared at the finish line of the plate, two yellow-green compounds appeared in the middle, and dark green compounds appeared at the start line, as shown in Figure 1. When comparing the coloration spots of both samples on the plate to the standard, it was determined that the absorbance at the 8th spot of the standard plate corresponds to ellagic acid.
The high-performance liquid chromatography (HPLC) analysis of ellagic acid was performed at wavelengths of 254.4 nm and 276.4 nm, using an isocratic eluent flow. According to the analysis results, the retention time of ellagic acid in the column was found to be between 10 and 11 minutes. The chromatogram obtained during the analysis is shown in Figure 3.
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The amount of pure compound in the ellagic acid mixture extracted from Maclura pomifera fruit was determined to be 88.18%. The analysis was repeated five times, and since no deviations were observed in the chromatogram, it was accepted as the specific
chromatogram for ellagic acid extracted from Maclura pomifera fruit. The key metrological parameters of the developed HPLC method for determining the amount of ellagic acid are presented in Table 2.
Table 2.
Metrological parameters for quantitative determination of ellagic acid
№ Compound Series Ellagic acid content in the obtained sample (%) Metrological Characteristics
1 12.10.2023 88,3 f=4 Sx=0,7 S=1,58 Хог=88,18 ЛХ=1,32 £o'r=3,99 % Р=0,01
2 17.11.2023 87,1
3 03.02.2024 88,9
4 07.04.2024 89,4
5 10.06.2024 87,2
The results of the metrological evaluation indicate that each sample of the mixture has a high level of purity, which enables the effective use of the extracted compound in further research and product development.
Conclusion. Based on the studies conducted on the extraction and chromatographic analysis of ellagic acid from Maclura pomifera fruit, it can be concluded that various methods have been used for ellagic acid
extraction, and the method we propose shows higher efficiency compared to other methods. The qualitative and quantitative analysis of the extracted ellagic acid was carried out using thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), and it was determined that the ellagic acid content in the extracted mixture is 88.18%.
References:
1. Gardner, E.M., Sarraf, P., Williams, E.W., & Zerega, N.J. C. (2017). Phylogeny and biogeography of Maclura (Mo-raceae) and the origin of an anachronistic fruit. Molecular Phylogenetics and Evolution, 117, 49-59. doi:10.1016/j.ympev.2017.06.021
2. Orazbekov, Y., Ibrahim, M.A., Mombekov, S., Srivedavyasasri, R., Datkhayev, U., Makhatov, B., Ross, S.A. (2018). Isolation and Biological Evaluation of Prenylated Flavonoids from Maclura pomifera. Evidence-Based Complementary and Alternative Medicine, 2018, 1-8. doi:10.1155/2018/1370368
3. Jones JM, Soderberg F: Cytotoxicity of lymphoid cells induced by Maclura pomifera (MP) lectin. Cell Immunol, 42, 319-326, 1979. DOI: 10.1016/0008-8749(79)90197-7
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ТЕХНИЧЕСКИЕ НАУКИ_январь. 2025 г.
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