DETERMINATION OF WATER SOLUBLE VITAMINS IN Ziziphora clinopodioides L. PLANT

ABSTRACT

This article describes the determination of water-soluble vitamins in the plant Ziziphora clinopodioides L. by High-Performance Liquid Chromatography (HPLC), information about the importance of the identified vitamins in the human body.

АННОТАЦИЯ

В статье описано определение водорастворимых витаминов в растении Ziziphora clinopodioides L. методом высокоэффективной жидкостной хроматографии (ВЭЖХ), приведены сведения о значении выявленных витаминов в организме человека.

Keywords: Ziziphora clinopodioides L., essential oils, flavanoids, macro and micronutrients, vitamins, food additive.

Ключевые слова: Ziziphora clinopodioides L., эфирные масла, флавоноиды, макро- и микроэлементы, витамины, пищевая добавка.

Introduction

Ziziphora clinopodioides L. is a perennial herb about 40 cm tall, belonging to the family of sedums. It blooms in June-July, bears fruits in July-August, grows on the slopes of the mountain district, on gravel and stony brown soil at an altitude of up to 2400 meters above sea level. Mainly the above-ground part of the plant (grass) is used. It is better than black pepper and laurel in its healing and pleasant taste. [1] This plant has been used in folk medicine since ancient times. It is found almost everywhere in the world. There are 7 species of it in our country, and it grows in mountainous regions of Tashkent, Samarkand, Jizzakh, Namangan, Kashkadarya and Surkhandarya regions [2]. Tincture of Ziziphora clinopodioides L. is used as an expectorant, anti-cold, analgesic, sedative, anthelmintic. One of the reasons for the widespread use of Ziziphora clinopodioides L. is its importance in the natural treatment of hypertension, which affects almost 60% of the population. Bod is an effective tool in the treatment of rheumatism, and in the elimination of flu viruses, which are considered to be highly contagious, and which flare up in the spring and autumn months [3].

Ziziphora clinopodioides L. contains triterpene, saponins, ursol, oleanol, chlorogen, flavonoids, coumarins [4], oxycoumarins [5], amino acids, macro and micro       elements (Ca, Si, Al, Fe, Mg, Co, Zn, Cu, Mn, Mo, Cr and selenium accumulates) [6,7]. In addition, Ziziphora clinopodioides L. contains essential oils [8], GC-MS analysis shows pulegone (5.19% - 57.85%), limonene (0.26% - 12.79%), 1,8-cineole (up to 27.4%), bornylacetate (0.47% - 9.37%), piperitone (0.70% - 9.05%) and menthol derivatives [9].

The aim of our research is to determine the vitamins contained in Ziziphora clinopodioides L. by the HPLC (high-performance liquid chromatography) method.

Experimental part

Acetate buffer system and acetonitrile were used as an eluent in the determination of water-soluble vitamins with the HPLC. Chromatographic conditions:

• Chromatograph Agilent-1200 (equipped with an autodoser)
• Column Exlipse XDB C 18 (obraschenno-faznyy), 5 μm, 4.6 x150mm
• Diode array detector (DAD), 204 nm, 254 nm, 290 nm identified.
• Selection of eluent (Flow rate 1ml/min., time, thermostat temperature 250^o  С, n=5, S=5 μl)

Table 1.

5-10 g of leaves of Ziziphora clinopodioides L. are taken on an analytical scale and placed in a 300 ml flat flask. It is extracted with 50 ml of 40% ethanol solution. The extract was heated under magnetic stirrer under reflux for 1 h with vigorous stirring and then stirred at room temperature for 2 h. The mixture is cooled and filtered. 25 ml of 40% ethanol was added to the remaining part and re-extracted 2 times. The filtrates were combined and filled to the mark with 40% ethanol (5-10%) in a 100 ml volumetric flask. The resulting solution is centrifuged at 7000 rpm for 10 minutes. The resulting solution was taken from the top for analysis. Working solutions of water-soluble vitamins with a concentration of 1 mg/ml were prepared. For this purpose, 50.0 mg of each vitamin standard is accurately drawn on an analytical balance and dissolved in 40% ethanol in a 50 ml volumetric flask and filled to the mark. First, working standard solutions and then prepared working solutions were introduced into the chromatograph. The obtained results are presented in Figure 1 and Table 2

Figure 1. Chromatogram of water-soluble vitamins in Ziziphora clinopodioides L. plant

Table 2.

Vitamins determined by the obtained chromatogram spectra

Results

Based on the above, vitamin B-9 dominates among water-soluble vitamins in Ziziphora clinopodioides L. plant. This vitamin is needed in the human body to maintain red and white blood cells (blood production), convert carbohydrates into energy (metabolism), and synthesize and maintain the body's genetic building blocks (DNA and RNA). In addition, it is necessary to ensure the healthy, normal development of the fetus and to prevent some serious birth defects.

Conclusion

Ziziphora clinopodioides L. plant contains water-soluble vitamins B₂, B₆, B₉, vitamin B₉ is the most abundant, and vitamin B₂ is the least. creation of food supplements made from used holly serves to satisfy people's need for vitamins, to strengthen their health and to extend their life.

References:

1. Askarov I.R. “Tabobat qomusi” . Tashkent “MUMTOZ SO‘Z” 2019.
2. I.F. Karimova, G.A. Salimova “Kiyik o‘t (zizifora)ning morfologiyasi, dorivorlik xususiyatlari va madaniylashtirish asoslari”. Scientific Journal Impact Factor 2021: 5.723
3. Askarov I.R. “Sirli tabobat”. Tashkent “Fan va texnologiyalar nashriyot-matbaa uyi” – 2021
4. K.Cmejkal, M.Malanik, K.Zhaparkulova, Z. Sakipova, L.Ibragimova, G. Ibadullaeva, M.Zemlicka. “Kazakh Ziziphora Species as Sources of Bioactive Substances” Molecules journal 2016, 21, 826
5. Franc¸ ois Senejouxa, C. Girarda, P. Kerramb, Haji Akber Aisac, A. Berthelota, Franc¸ oise Bévalota, C. Demougeota, “Mechanisms of vasorelaxation induced by Ziziphora clinopodioides Lam. (Lamiaceae) extract in rat thoracic aorta”. Journal of Ethnopharmacology · October 2010.
6. Kasymov F.U., Kyazimov I.M., Dembitsky A.D. and other. Компонентный состав эфирных масел видов Зизифора // Химия природных соединений. -1988.- № 23 (5) - Р. 636-637.
7. Woolf A. Essential oil poisoning // J Toxicol Clin Toxicol. - 1999. - №37(6). -P. 721-727.
8. Senejoux F., Demougeot C., Kerram P. et al. Bioassay-guided isolation of vasorelaxant compounds from Ziziphora clinopodioides Lam. (Lamiaceae)//Fitoterapia. - 2012. - №83. - P. 377-382.
9. Аghajani Z., Assadian F., Masoudi Sh et al. Cemical composition and In Vitro Antibacterial Activities of The Oil Ziziphora clinopodioides and Z. capitata SUBSP. capitata from Iran// ChemNat Comp. - 2008. - № 44(3) - P. 387-389.