GREEN INTENSIFIED EXTRACTION OF BIOACTIVE AND ANTIOXIDANT COMPOUNDS FROM TURNIP LEAVES

ABSTRACT

In this research, the chemical composition and antioxidant potential of Brassica rapa L. turnip leaves, specifically, its ecological and functional implications to health are investigated. The study provided an overview of botanical classification, bioactive compounds quantification, and methods followed in vitamins and phenolic compounds screening through HPLC determination. These findings supported that the leaves contained vitamin B₁ (6–7 mg/100g), vitamin PP (2–1 mg/100g), quercetin (5–32 mg/100g), and salicylic acid derivatives (2–31 mg/100g). DPPH assay indicated the detection of excellent antioxidant activity which was reported as IC₅₀ value 349.67 µg/mL, indicating positive antioxidant potential.

АННОТАЦИЯ

Исследователи изучили химический состав и антиоксидантную активность листьев репы Brassica rapa L. для их экологической и функциональной значимости для благополучия человека. Также был представлен обзор литературы о ботанической классификации, количественном содержании биологически активных соединений (в растениях) и методах идентификации витаминов и фенольных соединений с использованием анализа ВЭЖХ (высокоэффективная жидкостная хроматография). Из листьев были выделены витамины B₁ (6–7 мг/100 г), PP (1–2 мг/100 г), кверцетин (5–32 мг/100 г) и производные салициловой кислоты (2–31 мг/100 г). Метод DPPH антиоксидантной активности сильно проявляется при IC₅₀ 349,67 мкг/мл, что указывает на значительный антиоксидантный потенциал.

Keywords: Brassica rapa L., antioxidant activity, phenolic compounds, DPPH, eco-functional ingredients.

Ключевые слова: Brassica rapa L., антиоксидантная активность, фенольные соединения, DPPH, эко-функциональные ингредиенты.

Introduction

Turnip (Brassica rapa L.) is a plant grown globally and is a food and bioactive compound source. Several compounds of turnip or part of plant are reported in different studies [1; 345-405].

Guusje Bonnema-extra-budgetary pension fund under the Ministry of Finance of the Republic of Uzbekistan [2; 78-82]. Jianjun Zhao is a researcher of the Supreme Assembly of the Republic of Uzbekistan, who has done scientific work on the genetics and selection of turnip Camara-Martos F., Obregexn-Cano S., Mesa Plata O., Cartea-González ME, de Haro-Bailon A. (2020). Niccolo Bassetti is a member of the Legislative Assembly of the Republic of Uzbekistan, people's deputies on campaigning for regional, district and municipal councils (2015). Stefan Petrasch-events to the Legislative Chamber of the Supreme Assembly of the Republic of Uzbekistan, Regional, District and city councils of people's deputies (2022). Ningwen Zhang was the first sitting of the Legislative Assembly of the Republic of Uzbekistan. The work of these scientists was instrumental in increasing scientific knowledge of the genetics, biochemistry and physiology of turnip [3; 114–122].  

According to Ren YJ, Zhao ML, Han R. (2021) there are scientific works on turnip (Brassica rapa), but they are sufficiently advanced in relation to one path of information, while the possibilities for scientific research are still open to management. Several scientific researchers have explored the genetic diversity of turnip, Hosan, varieties and climate adaptation to study it [4; 14-18].

Complete information about the vitamins, minerals, and phytochemical composition of turnip (for example, glucosinolates and flavonoids) is publicly available. Turnip is frequently researched for its antioxidant, anti-flare, and immune-strengthening properties. Data for turnip diabetes, high blood pressure, and cardiovascular disease are provided by Gharibzahedi SMT and Jafari SM (2017) [5; 70–91].

Research on the effective use of turnip leaves, bark, and other protrusions is limited. There has been limited research on their utilization for bioactive substances or bioplastics, among other applications. Research on the creation of resistant varieties of turnip for construction and ball supply is not reliable. Few studies are available on the definition of turnip for anti-flare, anticorsinogen (cancer acquisition), and internal microflora. Turnip’s relevance as a biological pesticide, soil restoration, and ecological agriculture has not been examined widely. There is little research on the possibility of using natural extracts from turnips for medicinal or cosmetic purposes. Despite the extensive scientific research conducted on turnips, waste processing, extreme cooperative adaptation, medicinal properties, and other application options are still not available. These domains offer great potential for novel research opportunities [6; 127-130].

Materials and methods

Experience 1. The elimination Solutions of vitamins C (CAS 50-81-7), B₁ (CAS 59-43-8), B₆ (CAS 58-56-0), B₃ (CAS 59-67-6), B₁₂ (CAS 68-19-9), and PP (CAS 98-92-0) were prepared by dissolving 50 ml of 0.1 N solution of 5 mg of each vitamin (100 mg/l). Each standard solution of vitamins B₂ (CAS 83-88-5) and B₉ (CAS 59-30-3) was made by dissolving 5 mg of the aforementioned vitamins in a 50 ml 0.025% sodium hydroxide solution. The initial vitamins B₁, B₆, B₃, B₁₂, and B₉ were then removed from the vitamins by 200 µL, and a solution was prepared with a concentration of each vitamin of 14.286 mg/L. Thus, standard solutions of 7.143, 3.571, and 1.786 mg/l were prepared. Standard solutions of vitamin C with concentrations of 286, 143, 71.5, and 57.2 mg/l were prepared too. Pure water was used for a concentration of 0 mg/l to construct a calibrating graph in Table 1.

 Experience 2. Preparation of standard solutions. Gallic acid (5.2 mg), salicylic acid (5.2 mg), rutin (5 mg), quercetin (5 mg), apigenin (5 mg), and Kempferol (5 mg) were dissolved in 96% ethanol for 20 minutes in an ultrasonic bath before being transferred to a 50 ml flask and delivered to the line with ethanol. For each of these solutions, a total of 4 separate solutions were prepared (200 µL of each solution and mixed by peeling). After filling each solution, it was poured into the vial and analyzed. Plant extract preparation. For the phenolic compounds extraction, 1 g from the sample analyzed was sampled at an accuracy of 0.01 g on the NV222 brand scale produced by the OHAUS Company (USA), transferred to a conical flask with 50 ml volume, and 25 ml of 96% ethanol was added. The mixture was extracted for 20 min in a GT SONIC-D3 (Chinese) ultrasonic bath at 60°C.

Experience 3. A purple 2,2-diphenyl-1-picrylhydrazyl (DPPH) solution gets discolored, which makes it possible to identify a hydrogen atom or pure antioxidant compounds that give electron. Stable DPPH• is a reagent employed to perform spectrophotometric analysis. In the present study, the Blois approach as adapted by the DPPH• method for determining nature of free radical thinning was utilized with slight modifications. A 0.812 mM DPPH• solution was prepared with ethanol in a measuring flask with a volume of 100 ml, wrapped with aluminium paper and the solutions were stored in a dark place at room temperature for 30 minutes. A 4 ml quartz cuvette was placed in a spectrophotometer (3 ml of DPPH solution + 200 µL of ethanol  and the light absorption (D1) at 517 nm every 10 sec for 10 min was measured using the YOKE produced K7000 spectrophotometer (China). The antiradicality of the sample was established in the experiment by mixing 50, 100, 150, 200 µL of sample with 3 ml DPPH solution to determine light absorbency (D2) at 517 nm in the order presented above for each respective sample. Ethanol was added to the rest to bring the total volume of the solution in the cuvette to 3.2 ml. The antiradicality of the samples was determined by using (1) formula.

                                                            (1)

The results obtained were given in the following table (Table 1.).

Results and discussion

Vitamins  and reviews. Table 1 lists the total amount of vitamins left by 100 g of leaves calculated, based on the chromatogram of turnip leaf extract, as shown in figures 1-2. Figure 1. Determination of vitamins in the leaves of turnip leaf extract chromatogram. Table 1.

Table 1.

 Extract in turnip Leaf is the amount of vitamins and the time of Capture.

Table 1 shows that 100 g turnip Leaf was found to contain high amounts of vitamin B₁  67.3 mg, vitamin B₃  15.5 mg, vitamin PP 21.1 mg, and vitamin B 9  10.7 mg. Vitamin C was found to be 2.4 mg and vitamin B₆  was found to be 0.23 mg, Low.

Thiamine, a vitamin of Group B, has vital functions for Tashkent. Beriberi is the most frequent disease due to thiamine deficiency. Daily additions and desire are required to cause thiamine deficiency in the vitamin B₁ products. Vitamin B₁ is a vitamin that has been used to add and develop, and is responsible for cell function. Its main function is the conversion of energy, which recognizes the desired self-food. Other B vitamins and vitamin C are water-soluble vitamins. Resistant to hot temperatures. So the only reason Thiamine Reserve is missing in Tashkent is to take it from ordinary products. The target daily thiamine intake is 1.2 mg in men and 1.1 mg in women [7; 185–193].

Vitamin PP (also called niacin or vitamin B₃) is vitally important for the human body, playing vital roles in maintaining metabolic processes and general health. Vitamin PP is involved in metabolic processes within cells to ensure the release of energy from food. Niacin guards the nervous system and helps strengthen signal transmission processes. Helps in coping with Stress and mental conditions (depression and irritability). Supports the elasticity of blood vessels and protects it from atherosclerosis. Engages its key enzymes for digestion. Assists  the intestines to work more effectively. Assists skin cells to recover and maintains the skin’s health. The damage done by vitamin PP deficiency can be much bad. Sunlight  exposure skin redness and rash. Digestive  problems. Mental  disorder and memory loss. The daily level of vitamin PP is 6 - 12 mg in children, 14 mg in women, 16 mg in men, 18 mg in pregnant and 17 mg in lactating women. Daily requirement is mostly fulfilled with a balanced diet [8; 46-52].

2. Analysis of some phenolic compounds and results. A  chromatogram of a sample extract with weight (1 g) was drawn (Figure 3), and the amounts of phenol compounds in a sample 100 g were determined based on the analysis results with the following formula and quoted in Tables 2.

Figure 3. Chromatogram for the identification of polyphenols in the leaves of turnip leaf extract

Table 2.

Turnip leaf extract, the purpose and capture the Times of polyphenols.

Figure 3 gives the chromatogram for the polyphenol determination of the extract of turnip leaf and the amount and capture Times of polyphenols Table 2. This shows that the turnip Leaf contained 100 g of quercetin 532.6 mg, salicylic acid 230.9 mg, rutin 22 mg, apigenin 15.9 mg in larger quantity and no kaempferol and gallic acid were seen. But the influence of quercetin on human health and well-being is positive in every respect. It’s one of the antioxidants which are said to lower inflammation, increase cardiovascular health, and, some studies have found, can prevent cancer from developing. Quercetin can alleviate allergies and help the immune system.

But apigenin and other kinds of natural flavonoids appear in plants, including parsley, celery, chamomile and other greens. It has various health benefits, such as antioxidant, anti-inflammatory, anticancer. Apigenin protects cells from free radicals and oxidative stress. This process contributes to the slow down of cardiovascular disease and aging. Its role is to inhibit inflammatory mediators and consequently, chronic inflammatory diseases, including arthritis and asthma. Apigenin suppresses growth and proliferation of cancer cells. It’s especially active against breast, ovarian, prostate and even lung cancer. Apigenin does some neuroprotective things that lower the risk of nervous system disorders like Alzheimer's. It may also help to calming effect and enhance the quality of sleep. Apigenin modulates estrogen receptors in women to promote hormonal balance and diminish symptoms in menopause.

3.Determination of antiradical activity. To calculate the thinning concentration of the sample to 50% of IC50 – DPPH solution, the following graph was compiled in each experiment based on the values of 10-minute absorption (D2) and antiradicality activities (AA%) and calculated based on the trend line function transferred to it.

Figure 4. Brassica rapa L. leaf representation of measured light absorption of loose and examined sample solutions added to the DPPH solution

Figure 5. Brassica rapa L. leaf on the mesh of AA% s and volumes determined in the 10th minute of the sample

The trend line transferred to the graph was calculated from the function (2) formula y=mx+b to 50% AA% manifesting volume (IC50) X=(y-b)/M:

                                                     (2)

In place of the conclusion, it can be said that sample 1 exhibits antiradicality activity, in particular, its IC50 value was found to be 645.2 mcl.

Conclusion.

The current study aims to investigate the botanical classification, distribution, analysis and concentrations of the leaf of the variety Brassica rapa L. (turnip), a cultivar of the plant grown in Uzbekistan. State Standard requirements have been adopted by the Ministry of Health because parts of the plant have been consumed as food. Ash content, moisture content of essential nutrition (minerals, fiber content, vitamins, phenolic substances and antioxidants) should be considered. In terms of general health practice, using turnip leaves for nutrition is beneficial with rich phytochemicals that offer many benefits for the body, and is a part of a daily health diet. Then calcium and potassium appeared to be the most frequent minerals from those first annual leaves. Quercetin and apigenin were abundant in the total flavonoid content of leaves. The water-soluble vitamins stimulate and enhance vitamins, from other sources, that help to meet the natural requirements. It is readily accepted and is well used in food, natural storage without artificial additives, adding of the turnip leaf to functional products to reduce the spoilage of the production of new products. Instead of summarising, we can just eat Brassica rapa L. This plant contains the leaf rather than the fruit part, scientifically according to sources. Vitamins and flavonoids are believed to be highly radical resistant. Since Brassica rapa L. has antiradical activity, we assessed the antiradical activity of Brassica rapa L. using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method for leaf. According to the observation obtained from 50% of the IC50 – DPPH solution, the leaf concentration after 10 min was determined as 50 to 200 mcl and values for antiradical activities are calculated for Brassica rapa L. Therefore, antiradical activity was found on leaf level in the radishes grown in Uzbekistan Brassica rapa L. therefore IC50 is equal to 645.2 mcl. Therefore, the antiradical activity of Brassica rapa L. indicates that many diseases such as prevention and treatment disorders are related to the health of humans.

References:

1. Abdul-Razzaq has sholgom (Brassica rapa var. rapa L.) breeding. In: Al-Khairy JM, Jain SM, Johnson DV, editors. Advances in plant-growing strategies: vegetable crops. Springer international; Cham, Switzerland: 2021. Pp. 345-405. Volume 8: bulb, root, and root. [DOI] [Google Scholar].
2. Asqarov I. R., Muminzhonov M. M., Atakulova N. B.. Chemical composition and medical properties of watermelon fruit. Scientific newsletter.- Fergana, 2022. -1.78-82B. site: www.fdu.uz.
3. Asqarov I. R., Atakulova N. B., Anvarova M. M. Plasma index garden determination of macro-and microelements of watermelon wars of the "sweet" variety by the method of mass spectrometry (ISP-MS). Journal of commodity chemistry and traditional medicine 2022. 1(5), 114–122. https://doi.org/10.55475/jcgtm/vol1.iss5.2022.108.
4. Bonnema G., Li JG, Shuhang W., Lagarrigue D., Bucher J., Wehrens R., de Vos R., Beekwilder J. glucosinolate variability between turnip organs during development. PLoS ONE. 2019; 14-18. e0217862. doi: 10.1371 / journal.pone.0217862. [DOI] [PMC free article ] [PubMed] [Google Scholar].
5. Mitreiter S., Gigolashvili T. regulation of glucosinolate biosynthesis. J. Exp. Bot. 2021;72: 70–91. doi: 10.1093 / jxb / eraa479. [DOI] [PubMed] [Google Scholar].
6. Camara-Martos F., Obregexcessn-Cano S., Mesa Plata O., Cartea-Gonzexcesslez ME, de Haro-Bailon A. Brassicaceae the amount of glucosinolate and microelements in leafy vegetables and bioavailability in vitro. Esophageal Chemistry. 2021; 339: 127860. doi: 10.1016/j. foodchem. 2020. 127-130. [DOI] [PubMed] [Google Scholar].
7. Thiruvengadam M., Chung IM.   Selenium, putresine, and cadmium are phytochemicals and turnips that strengthen salinity (Brassica rapa ssp. rapa) in molecular daroja gives a definition of esophageal chemistry. 2015;173:185–193. doi: 10.1016 / j. foodchem.2014.10.012. [DOI] [PubMed] [Google Scholar].
8. Rana A., Samtia M., Dhewa T., Mishra V., polyphenols Aluko back health care tasks: a brief review. J. esophageal biochemistry. 2022; 46-52. e14264. doi: 10.1111 / jfbc.14264. [DOI] [PubMed] [Google Scholar].