بررسی و مقایسه ماده مؤثره اپی گالوکاتچین گالات در کالوس حاصل از کشت بافت گیاه دارویی (Camellia sinensis (L.) Kuntze) به روش نوین استخراج با پلیمر قالب مولکولی (MIP)
محورهای موضوعی : فیتوشیمیآذرم موحدی 1 , مازیار احمدی گلسفیدی 2 , مهدی علیزاده 3
1 - گروه زیست شناسی- دانشکده علوم پایه- واحد گرگان- دانشگاه آزاد اسلامی- گرگان- ایران
2 - معاون پژوهش و فناوری- دانشگاه آزاد اسلامی گرگان
3 - عضو هیئت علمی گروه علوم باغبانی/ دانشگاه علوم کشاورزی و منابع طبیعی گرگان
کلید واژه: کشت بافت, چای سبز, کالوس, پلیمر قالب مولکولی, اپی‌گالوکاتچین گالات,
چکیده مقاله :
به دلیل اهمیت اپی گالو کاتچین گالات EGCG” “در گیاه چای در صنایع دارویی و غذایی، به بررسی و مقایسه میزان EGCG در عصاره برگ جوان (دوبرگی) و مسن و کالوس گیاه پرداخته و اقدام به جداسازی و اندازه‌گیری EGCG در عصاره کالوس به روش پلیمر قالب مولکولی MIP در سال 1397در داروسازی گیاه اسانس شد. جهت تولید و تحریک کال‌زایی، کالوس گیاه چای برداشت شده از منطقه غرب مازندران در محیط‌های کشت MS، SH و WPM با مقادیر مختلف از هورمون‌های گیاهی کشت شد. برگ و کالوس با متانول 70 درصد به روش ماسراسیون عصاره-گیری شدند. برای خالص سازی EGCG از عصاره کالوس، مقدار نانولوله‌ی چند دیواره و عامل‌دار کربن (MWCNTs) در محدوده‌ی 10-1 میلی‌گرم، در ساخت CNTs-MIP بررسی و بهینه شد. EGCG در عصاره کالوس با CNTs-MIP ، استخراج و میزان آن با کروماتوگرافی فاز مایع تعیین شد. بیشترین حجم کالوس در محیط WPM و بیشترین میزان EGCG در محیط SH با غلظت بهینه از هورمون‌های BA (2 mg/L), 2,4-D (0.5 mg/L)، مشاهده شد. افزودن 8 میلی‌گرم از MWCNTs به ساختار پلیمر، میزان بیشتری از EGCG را استخراج نمود. EGCG در برگ جوان و مسن و کالوس گیاه به ترتیب 34/37، 19/7، 13/0 میلی‌گرم در گرم می‌باشد. میزان EGCG در کالوس با استفاده از CNTs-MIP، 91/0 میلی‌گرم در گرم محاسبه شد. با وجود مقدار کم EGCG در کالوس، با استفاده از CNTs-MIP مقدار این ماده با ضریب تغلیظ 8 برابری قابل اندازه‌گیری و بیانگر کارایی این تکنیک می باشد.
Due to the importance of constituent Epigallocatechin gallate (EGCG), in the tea plant in use of pharmaceutical and food industries, the amount of EGCG in young leaf, old leaf and callus extract was compared and measured. EGCG in callus extract was isolated and measured by Molecularly Imprinted Polymer (MIP) technique in 1397 in Giah Essence Phytopharm Company. In order to callus induction, callus of tea plant which was harvested from the western region of Mazandaran province cultured in WPM, SH, MS media with different amounts of plant hormones. Leaves and callus extracted with 70% methanol by maceration method. For optimization of extraction and purification, various quantities of functional multiwalled-CNTs 1-10 mg used in production of CNTs-MIP and the performance investigated. EGCG of callus extract extracted with CNTs-MIP and its amount determined with HPLC. The highest volume of callus in WPM medium ,and the highest amount of EGCG in SH medium observed with optimized concentrations of hormones BA(2 mg/l), 2,4_D(0.5 mg/l). The highest content of EGCG was extracted by adding 8 mg of MWCNTs in the polymer structure. EGCG in young and old leaves and callus were 37.34, 7.19 and 0.13 mg/g of dried weight, respectively. By CNTs-MIP, the EGCG content in callus was obtained 0.91 mg/g. Despite the trace amount of EGCG in callus, the amount of this substance can be measured with a concentration coefficient of 8 times by CNTs-MIP, demonstrated the efficiency of this technique.
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