|Year : 2018 | Volume
| Issue : 2 | Page : 278-281
Novel eco-friendly “One-Pot” facile strategy for production of the natural quercetin from the plant: A model study
Elena Vladimirovna Maksimenko, Anna Vladimirovna Lekar, Salima Salimovna Khizrieva, Sergey Nikolaevich Borisenko, Elena Vladimirovna Vetrova, Nikolay Ivanovich Borisenko, Vladimir Isaakovich Minkin
Laboratory of Critical Fluid Technologies, Institute of Physical and Organic Chemistry, Southern Federal University, Stachky Avenue 194/2, Rostov-on-Don, Russian Federation, Russia
|Date of Web Publication||20-Jun-2018|
Nikolay Ivanovich Borisenko
Institute of Physical and Organic Chemistry of Southern Federal University, Stachky Ave., 194/2, Rostov-on-Don 344090
Source of Support: None, Conflict of Interest: None
| Abstract|| |
This work is aimed at the development of an “one-pot” technique for the production of the natural antioxidant quercetin (QR) from the buds of Sophora Japanese (Sophora japonica L.) using the subcritical water (SBW). For the first time, SBW that serves as a reactant and a solvent has been used to obtain QR in good yields starting from the buds of Sophora Japanese. High-performance liquid chromatography was used to determine the quantitative and qualitative compositions of the obtained products. For the first time, a new eco-friendly “one-pot” technique was proposed for production of QR from the buds of Sophora Japanese using SBW. This method of preparation of QR allows one to avoid the use of toxic organic solvents. The good yields of the targeted QR can be obtained for the period of time which is ten times shorter than needed by the traditional procedures.
Keywords: Flowers buds, one-pot, quercetin, Sophora Japanese, subcritical water Introduction
|How to cite this article:|
Maksimenko EV, Lekar AV, Khizrieva SS, Borisenko SN, Vetrova EV, Borisenko NI, Minkin VI. Novel eco-friendly “One-Pot” facile strategy for production of the natural quercetin from the plant: A model study. J Nat Sc Biol Med 2018;9:278-81
|How to cite this URL:|
Maksimenko EV, Lekar AV, Khizrieva SS, Borisenko SN, Vetrova EV, Borisenko NI, Minkin VI. Novel eco-friendly “One-Pot” facile strategy for production of the natural quercetin from the plant: A model study. J Nat Sc Biol Med [serial online] 2018 [cited 2021 Mar 8];9:278-81. Available from: http://www.jnsbm.org/text.asp?2018/9/2/278/234702
Quercetin (QR) (3,3 ', 4', 5,6-Pentahydroxyflavone) is a plant polyphenol containing in many fruits, vegetables, leaves, and grains. It relates to the vast family of flavonoids which are secondary metabolites in plants found multitude applications in the pharmaceutical, nutritional, and cosmetic industries because of their useful biological activities, particularly strong antioxidant, chelation, anticarcinogenic, cardioprotective, bacteriostatic, and secretory properties., This makes QR beneficial to human health and leads to constantly increasing interest in the use of QR as a dietary supplement to various traditional food products. The recent studies of the biological properties of QR have shown that it helps to reduce growing senescent cells in vivo and serves as an efficient senolytic drug.,
The widespread use of QR in the food industry and the promising prospects for application in medicine has already aroused considerable interest in the elaboration of low-cost and eco-friendly technologies for its production. At present, the conventional procedure for preparation of QR as a pharmacological substance or a dietary addition includes two basic steps [Figure 1]. The first one is associated with extraction of QR-containing glycosides (rutin (RT), rutin-7-O rhamnoside) from various plants and their subsequent purification. The second step involves the procedure of the hydrolysis of the purified QR glycosides to obtain the QR and perform its purification. These processes, as a rule, are separated in time and space.
|Figure 1: The principal reaction schemes for preparation of quercetin from flower buds of Sophora Japanese|
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Currently, within the framework of the concept of “green chemistry,” an avalanche-like growth is observed of the number of studies devoted to the development of “a one-pot synthesis” techniques and procedures. In chemistry, “a one-pot synthesis” is a strategy to improve the efficiency of a chemical reaction, whereby a reactant is subjected to successive chemical reactions in just one reactor. This approach is much greeted by chemists because it allows avoiding a lengthy separation process and purification of the intermediate chemical compounds, saves time and resources, and not infrequently increases chemical yield. It is thus interesting to apply the “one-pot” strategy for numerous tasks related to the extraction and transformation of plant's secondary metabolites. The successive implementation of this strategy could open up a perspective for the future development of inexpensive and environmentally friendly technologies to the production of new plant-based substances for pharmaceutical, nutritional, and cosmetic industries.
In the present work, as a model source of flavonoids, we used flower buds of Japanese Sophora flowers. Sophora Japanese (Sophora japonica L.) is often used as a plant material for the production of flavonoids and, in particular, rutin [Figure 1] from the legume family, which is widespread in Asia and cultivated in Southern regions of Russia. Leaves, flowers, and flower buds of Japanese Sophora are widely employed in traditional Chinese medicine because they contain a wide range of biologically active compounds: flavone glycosides, isoflavones, chromones of coumarins, saponins, triterpene glycosides, phospholipids, alkaloids, amino acids, polysaccharides, and fatty acids. For the extraction of flavonoids from the plant matrix, including buds of Sophora, various two-step methods  have been developed.
As already noted above, the two basic steps are required to obtain of the natural antioxidant QR from plants, including the step of extracting QR glycosides (for example, rutin) and subsequent hydrolysis thereof. Typically, traditional extraction of buds of Sophora is carried out using various organic solvents. The most common technique is the extraction with 70%–95% ethanol usually taken from 20 min to 4 h  and affording rutin in the yields from 0.2% to 18%. Also common is extraction with methanol occurring under reflux  and requiring infusion for 48 h  or 18 h with preliminary shaking for 6 h.
The process can be accelerated by the ultrasound irradiation. The yields of rutin in the methanol extracts are higher (15%–18%) than those achieved in the ethanol extraction procedures. The conventional synthesis of QR from the plant involves the acidic hydrolysis of the glycosides usually catalyzed by mineral acids (phosphate, sulfate, and hydrochloric acids). The reactions are lasting –3 h. Often used is also enzymatic hydrolysis of (reaction time [RT] is in the range of 12–24 h)., It should be noted that conventional acid hydrolysis of RT requires 2–3 h. The products of the hydrolysis are needed of careful purification. Thus, the conventional methods of preparation of RT require significant time costs.
An alternative to the conventional methods based on using acids to catalyze hydrolysis is to perform the reactions in SBW. In recent years, SBW has been used as a cheap, environmentally friendly solvent for extraction, synthetic transformations, and the recycling of different organic wastes. The hydrothermal reaction has been attracting much attention because of the fascinating physical and chemical characteristics of water near its critical point. In these conditions, water exhibits a much lower dielectric constant and a much larger ion product. The ion product or dissociation constant is about 3 orders of magnitude higher near the critical point (in the temperature, range from 220°C to 270°C) than it is for ambient liquid water. Under these conditions, there is a high H3O + and OH − ion concentration. As such, some acid-catalyzed organic reactions can be carried out without the addition of acids. However, the ion product decreases greatly above the critical point. This fact makes subcritical water an ideal reaction medium for the hydrolysis of organic compounds.
In this study, it is for the first time proposed to perform extraction of QR glycosides simultaneously with the subsequent hydrolysis as a “one-pot” process in a medium of subcritical water (SBW). This approach makes possible to avoid the use of flammable and costly, toxic organic solvents. In this regard, the purpose of the presented paper was the development of an expedient eco-friendly procedure for producing of QR in good yields starting from flowers buds of Sophora Japanese (S. japonica L.).
| Materials and Methods|| |
The flowers buds of S. japonica were purchased from JSC “Azbuka trav” (Russia). Acetonitrile (high-performance liquid chromatography [HPLC] grade) and water for the mass -spectrometry were purchased from JSC “Vekton” (Russia). Rutin (C27H30O16, Mw 610.25 ≥96.6%) was purchased from “Merk.” QR (C15H10O7, Mw 302.10, ≥98.2%) was purchased from JSC “Diaem” (Russia).
HPLC analysis was performed using a Agilent 1200 LC system, including a quaternary pump, a temperature controlled column compartment, an autosampler, and a diode-array detector. Eclipse XDB reversed-phase column C8 150 mm × 2.1 mm, 3.5 μm, was used for HPLC analysis. The mobile phase composition was as follows: CH3 CN: 0.5% H3 PO4, 78:22; column temperature 30°C; mobile phase flow rate, 0.14 mL/min; ultraviolet-detector wavelengths, 360 nm.
The conventional way for preparation of QR from the buds of S. japonica included two basic steps. The step one is to obtain of rutin from the flower buds through extraction with using the organic solvents and subsequent purification. The step two involves the procedure of hydrolysis of the extraction of rutin that obtained from the flower buds, to release the QR using the organic solvents and the mineral acids and subsequent purification. The extraction of rutin from the buds of S. japonica included the several stages. A 1 g sample of the dry buds of S. japonica (particle size: 0.5–1.0 mm) was boiled four times under reflux. At the first stage, dry buds were boiled with hexane 30 ml for 90 min. At the next three stages, 30 mL of 80% aqueous solution of ethanol was boiled for 90 min. The obtained extracts were filtered, combined, and analyzed by HPLC.
The hydrolysis of RT from extract of S. japonica buds was performed using hydrochloric acid. A sample of 0.10 g of extract (content of RT 38.6%) was dissolved in 2 ml of 80% methanol and 0.25 ml of concentrated hydrochloric acid (density 1.179) is added. The hydrolysis is carried out for 45 min at 100°C. At the end of the hydrolysis, the obtained precipitate is filtered through paper and washed with distilled water to a neutral pH. The washed precipitate is dried at 80°C for 2–3 h; the weight of the precipitate is 0.0268 g. Quantity of the QR in precipitate is 55.6%.
The “one-pot” way for preparation of QR from the buds of S. japonica by SBW, development in this paper, included the one single step: treatment of the flower buds using the medium of SBW.
The treatment's procedure of the flower buds in the SBW was performed using self-made reactor (autoclave). The reactor has inner volume 10 mL. The buds (0.1 g) were put into self-made stainless steel reactor. Into reactor was filled with 7 mL of the water's solution of sulfuric acid (0.25% ). The reactor was hermetically closed and put into a drying oven, where it was kept at a certain temperature (accuracy ± 1°C) for 30 min. After that, the reactor was cooled down to room temperature (15 min) in a tank filled with cold water. Its content was quantitatively transferred into a paper filter, filtered, and washed with distilled water to a neutral pH and after that washed with 80% EtOH until the color disappears. The aliquots of the solution obtained were diluted to the concentration required for analysis by HPLC/MS.
| Results and Discussion|| |
In accordance with the tasks of this study, the products containing QR were obtained from flowers buds of Sophora Japanese using two different schemes [Figure 1]. The scheme 1 conforms to the conventional two-step way of preparation of QR and includes traditional extraction by ethanol following by the hydrolysis of the obtained extract with HCl as the catalyst. The second scheme pictures the “one-pot” production of QR with the use of SBW. The obtained targeted products were analyzed for the contents of QR.
At the first stage, the amounts of RT and QR contained in the test sample were determined using a conventional extraction by ethanol. It was found that 1 g of the used raw material of the flower buds contains 191.1 mg of RT and 4.8 mg of QR. After treatment of the extract obtained by the two-step procedure with the use of HCl, the content of QR was increased to 91.1 mg/g.
At the next stage, the yields of QR were studied using SBW for realization of the “one-pot” technique. Our previous studies showed that the conversion of RT into QR, using the SBW medium, was most complete at the temperatures range of 100°C–250°C., Therefore, here, we studied in detail the temperature dependence of the yield of the RT and QR in medium of SBW in this temperature range. Also studied was the composition of the products obtained by the conventional procedure, starting from the flower buds of Sophora Japanese.
The dependence of the amount of RT and QR in the products obtained from flower buds of Sophora Japanese in SBW (without any additives) demonstrates an increase in the yield of QR (from 0.6 to 27.3 mg/g) when elevating the temperature to 200°C and corresponding decrease in the yield of RT. The decrease in the amount of RT, as was previously shown, is caused by the hydrolysis processes in which SBW serves as the catalyst. The further increase in temperature led to a decrease in the amount of QR. Analysis of the data obtained showed that at the temperatures range of 200°C–210°C, the yield of QR was the highest but still lower than the theoretical yield (if the stoichiometry of hydrolysis is taken into account in this case). At a temperature of 230°C, RT was not detected by HPLC in either the precipitate or solution. Hence, the hydrolysis was complete, and the decrease in the yield of QR at the temperature above 210°C is caused by its thermal destruction. The QR yield in the temperature range of 220°C–250°C does not exceed 30 mg/g, whereas they should grow to 90 mg/g. It can be assumed that an increase in temperature above 210°C leads to a significant decrease in the yield of QR due to its thermal destruction. The results obtained are in accord with the previous data obtained in the study of the conversion of rutin to QR in SBW.,
These data allow selecting the temperature of 200°C as the temperature at which the effect of the thermal degradation of QR on its yield is negligible. However, since the dissociation constant of SBW reaches a maximum in the higher temperature (between 220°C and 270°C), the acidity of SBW at 200°C is insufficient for complete hydrolysis of RT. Therefore, we assumed that increase in the yield can be achieved with the use of catalytic amounts of strong mineral acids. To determine the effect of trace amounts of acids, the dependence of the QR yield from buds of Sophora on the acid concentration (H2 SO4) and the processing time (in the interval of time from 10 to 60 min) was studied [Figure 2].
|Figure 2: The yields of reaction time and quercetin starting from 1 g of the flower buds of Sophora Japanese (mg). Conditions: (1) Subcritical water 200°C, t = 60 min, (2) subcritical water 200°C +0.1% H+, t = 60 min, (3) subcritical water 200°C +0.1% H+, t = 30 min, (4) subcritical water 200°C +0.1% H+, t = 20 min, (5) subcritical water 200°C +0.25% H+, t = 10 min (6) subcritical water 200°C + 0.25% H+, t = 30 min, (7) subcritical water 200°C +0.25% H+, t = 40 min, (8) Two-stage conventional procedure|
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As can be seen from the data pictured in [Figure 2], the use of trace amounts of acid made it possible to achieve the yield of QR obtained by the proposed “one-pot” technique comparable to that obtained by the employment of conventional techniques at the significantly (by 10 times) decreased temporary costs. The important advantage of the proposed procedure free of the use of toxic organic solvents is that it allows preparation of the final products in sufficiently pure state which excludes an additional stage of their purification. The comparison of the effectiveness of the developed and traditional methods is given in [Table 1].
|Table 1: Comparison of the effectiveness of the different techniques for preparing quercetin from flower buds of Sophora Japanese (sample 1 g)|
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| Conclusions|| |
A novel eco-friendly “one-pot” facile technique was proposed for the preparation of the natural antioxidant QR from flower buds of S. japonica using SBW.
The method way requires no use of toxic and flammable organic solvents. The suggested procedure is significantly (by 10 times) faster than the commonly used conventional techniques.
The proposed technique has a potential for the future development of low cost and enviromentally friendly technologies for the production of the new plant-based substances for the pharmaceutical, nutritional, and cosmetic industries.
This research was supported by the Internal Grant of the Southern Federal University (Project No BнГp-07/2017-04).
Financial support and sponsorship
This research was supported by the internal grant of the Southern Federal University (Project No BнГp-07/2017-04).
Conflicts of interest
There are no conflicts of interest.
| References|| |
Hirpara KV, Aggarwal P, Mukherjee AJ, Joshi N, Burman AC. Quercetin and its derivatives: Synthesis, pharmacological uses with special emphasis on anti-tumor properties and prodrug with enhanced bio-availability. Anticancer Agents Med Chem 2009;9:138-61.
Rashid Q, Azhar M, Naseem A, Khan M, Singh P. The specificity of non-heparin based conformational activators of antithrombin for factor Xa inhibition. J Nat Sci Biol Med 2011;2:49.
Zhu Y, Tchkonia T, Pirtskhalava T, Gower AC, Ding H, Giorgadze N, et al.
The Achilles' heel of senescent cells: From transcriptome to senolytic drugs. Aging Cell 2015;14:644-58.
He X, Bai Y, Zhao Z, Wang X, Fang J, Huang L, et al.
Local and traditional uses, phytochemistry, and pharmacology of Sophora japonica L.: A review. J Ethnopharmacol 2016;187:160-82.
Lekar AV, Filonova OV, Borisenko SN, Maksimenko EV, Vetrova EV, Borisenko NI,et al
. Extraction of bioflavonoid mixture from onion husk by subcritical water. Russ J Phys Chem B 2013;7:829-36.
Mihaylova D, Schalow S. Antioxidant and stabilization activity of a quercetin-containing flavonoid extract obtained from Bulgarian Sophora japonica L. Braz. Arch Biol Technol 2013;56:431-8.
Xu J, Zhang H, Cheng G. Sophora flowers (Flos Sophorae Immaturus). Talanta 2007;73:932-7.
Hermans-Lokkerbol A, Ingkaninan K. Separation and detection of the antioxidant flavonoids, rutin and quercetin, using HPLC coupled on-line with colorimetric detection of antioxidant activity nitra nuengchamnong. Naresuan Univ J 2004;12:25-37.
Velikorodov AV, Shevtsova IA, Fedorovich VV. Study of chemical structure of fruits and inflorescences of Chinese scholar tree, introduced in the Astrakhan region. Exp Chem 2010;2:164-9.
Paniwnyk L, Beaufoy E, Lorimer JP, Mason TJ. The extraction of rutin from flower buds of Sophora Japonica. Ultrason Sonochem 2001;8:299-301.
Graefe EU, Wittig J, Mueller S, Riethling AK, Uehleke B, Drewelow B, et al.
Pharmacokinetics and bioavailability of quercetin glycosides in humans. J Clin Pharmacol 2001;41:492-9.
Wang J, Zhao LL, Sun GX, Ling Y, Wu FA, Chen Z, et al
. A comparison of acidic and enzymatic hydrolysis of rutin. Afr J Biotechnol 2011;10:1460-6.
Galkin AA, Lunin VV. Subcritical and supercritical water: A universal medium for chemical reactions. Russ Chem Rev 2005;74:21-35.
Vetrova EV, Maksimenko EV, Khizrieva SS, Bugaeva AF, Borisenko NI, Minkin VI. Asimple way for the preparation of natural antioxidant quercetin from rutin by subcritical water. J Nat Sci Biol Med 2017;8:213-5.
[Figure 1], [Figure 2]