ارزیابی تاثیر ترکیب حلال و کاربرد تیمارهای کمکی در استخراج عصاره زرین‌گیاه (dracocephalum kotschyi boiss) به کمک حلال‌های یوتکتیک عمیق

در این مطالعه، استخراج ترکیبات موثره زرین‌گیاه با استفاده از حلال‌های یوتکتیک در سه مرحله انجام شد. در مرحله اول، تاثیر ترکیبات مختلف حلال شامل کربوکسیلیک‌اسیدها (سیتریک‌اسید و مالییک‌اسید)، قندها (گلوکز و فروکتوز) و قندالکل‌ها (اتیلن‌گلیکول و گلیسرول) بر کیفیت عصاره استخراج‌شده ارزیابی شد. نسبت ترکیبات گیرنده پیوند هیدروژنی به اهداکننده پیوند هیدروژنی (کولین کلراید) 1:1 (مولار) درنظر گرفته شد. در مرحله دوم، به‌منظور بهبود استخراج عصاره، نمونه تحت تیمار با امواج الکتریکی پالسی در دو شدت میدان 2000 ولت و 5000 ولت قرار گرفت و در مرحله سوم استخراج ترکیبات با حلال منتخب از مرحله اول در شرایط اعمال و عدم اعمال امواج فراصوت در صفر، 15 و 30 دقیقه تحت اعمال امواج فراصوت با توان 200 وات صورت پذیرفت. نتایج نشان داد که عصاره مستخرج با حلال حاوی اتیلن‌گلیکول به‌دلیل بالا بودن فعالیت آنتی‌اکسیدانی و عصاره مستخرج با حلال حاوی مالییک‌اسید به‌دلیل بالا بودن ترکیبات فنولیک به‌عنوان بهترین حلال‌ها انتخاب شدند. محتوای ترکیبات موثره عصاره مستخرج با حلال اول 69.5 % بیشتر از حلال دوم ارزیابی شد و در این مرحله حلال یوتکتیک حاوی کولین‌کلراید-اتیلن‌گلیکول برای استخراج عصاره زرین‌گیاه انتخاب شد. با افزایش ولتاژ امواج الکتریکی پالسی بر نمونه و مدت زمان اعمال امواج فراصوت در مرحله استخراج، محتوای ترکیبات فنولیک استخراج شده آن افزایش یافت. مجموع وزنی ترکیبات شناسایی شده درعصاره مستخرج با حلال یوتکتیک حاوی اتیلن‌گلیکول با اعمال ‌تیمارهای کمکی 25275.41 و بدون اعمال ‌تیمارهای کمکی 17000.05 میکروگرم بر گرم ارزیابی شد. اعمال پیش‌تیمار امواج الکتریکی پالسی 5000 ولت و استفاده از امواج فراصوت به‌مدت 30 دقیقه در مرحله استخراج، تاثیر معنی‌داری در افزایش قابلیت استخراج ترکیبات موثره زرین‌گیاه نشان دادند. با استفاده از روش رسوب ضد حلال، 53% کل ترکیبات فنولیک استخراج شده بازیابی و حلال یوتکتیک به سیستم بازگردانده شد.

effect of solvent composition and auxiliary treatments on the extraction of dracocephalum kotschyi boiss extract using deep eutectic solvents

introductionrecovery of active ingredients from plants is generally carried out using solid-liquid extraction. selecting an appropriate solvent is one of the parameters that strongly affects the extraction performance and the type of extracted compounds. so far, various solvents have been used to extract these compounds. in addition to the need for large amounts of solvent, the high consumption of organic solvents causes problems such as environmental pollution and possible destruction of active compounds. green solvents have been considered for the extraction process to reduce the consumption of non-toxic solvents and protect the environment. natural deep eutectic solvents (nades) are a new class of solvents used in extraction that consist of a mixture of two or more biodegradable natural compounds with low or no toxicity. the important features of these solvents are their low toxicity, biocompatibility, simple preparation, and low cost. given the significant advantages of these solvents, in recent years, the use of nades in the extraction of phenolic and antioxidant compounds has been considered. applying appropriate auxiliary treatments to the sample or the sample-solvent combination during the extraction process can improve the performance of the extraction process. the use of ultrasonic waves allows for better extraction by creating shear force, disrupting the integrity of the cell wall, and better penetration of the solvent into the tissue. pulsed electric waves also create pores in the cell membrane without causing minimal damage to the other parts, while maintaining consistency and structure, accelerating the permeability and transport of water and dissolved substances from the cell membrane, and allowing the extraction of active substances under better conditions.materials and methodsthis study was conducted in 3 phases to extract the compounds of the golden plant using natural deep eutectic solvents. in the first phase, effect of the type of solvent used including carboxylic acids (citric acid and malic acid), sugars (glucose and fructose), and sugar alcohols (ethylene glycol and glycerol) on the quality of the extracted material was evaluated. in the second phase, to improve the possibility of extracting the active ingredient of this plant, the sample was treated with pulsed electric waves, and in the third phase, the extraction of phenolic compounds from the sample was carried out with the solvent type selected from the first phase under the conditions of applying and non applying ultrasonic waves. identification of the compounds present in the extract was carried out on the selected samples using hplc. finally, the ability to recover the extracted compounds was carried out from the best sample. statistical analysis of the results was carried out using a completely randomized design - factorial test with sas version 9 software.results and discussionthe extract from the eutectic solvent containing ethylene glycol due to its high antioxidant activity and the extract from the eutectic solvent containing maleic acid due to its high phenolic compounds were selected as the best solvents for making the dracocephalum kotschyi extract. considering the total number of identified compounds, the content of identified compounds in the extract with a eutectic solvent containing ethylene glycol was 17000.05 μg/g, and in the extract with a eutectic solvent containing maleic acid was 10029.1 μg/g. therefore, the content of active compounds of an extract with a eutectic solvent containing ethylene glycol was about 70% higher than the content of active compounds of an extract with a eutectic solvent containing maleic acid, and this solvent was selected for further studies. the study of the effect of applying electric pulses and ultrasonic waves on the extraction process shows that by increasing the intensity of the electric pulse and the duration of using ultrasonic waves, the content of phenolic compounds and antioxidant properties of the extract increased. electric pulses accelerate the permeability and transport of water and dissolved substances by creating pores in the cell membrane. the change in the properties of the cell wall membrane in such a way that the substances inside the cell can be quickly and easily removed from the cell, causes the extraction to be carried out in the minimum time and energy required. the total weight of phenolic compounds identified in the extract with a eutectic solvent containing ethylene glycol with the application of auxiliary was 25275.41 μg/g, without the application of auxiliary treatments was 17000.05 μg/g, and in the sample extracted with ethanol was 21652.89 μg/g. therefore, the application of auxiliary treatments was effective and superior to the ethanol solvent in increasing the extraction of compounds from the plant. the polyphenol content extracted with ethylene glycol-choline chloride with and without auxiliary treatments was determined as 53 and 45 percent, respectively.conclusionthe solvent containing choline chloride-ethylene glycol had the best conditions for extracting the active compounds of dracocephalum kotschyi. applying a 5000 w electric pulse pretreatment and using ultrasonic waves for 30 minutes in the extraction stage had a significant effect in increasing the extractability of the active compounds. using the anti-solvent (water) precipitation method, 53% of the phenolic compounds were recovered and the eutectic solvent was returned to the system.