Optimization of total saponin extraction from tam that roots using the desirablity methodology

In Vietnam, Tam that hoang (Panax stipuleanatus Tsai et Feng) belonging to Araliaceous family is a valuable medicinal plant that possesses many biologically active compounds. Among them, flavonoids, polysaccharides, and saponins are the major constituents of this plant.
Nội dung trích xuất từ tài liệu:
Optimization of total saponin extraction from tam that roots using the desirablity methodologyJOURNAL OF SCIENCE OF HNUE DOI: 10.18173/2354-1059.2016-0052Natural Sci. 2016, Vol. 61, No. 9, pp. 26-33This paper is available online at http://stdb.hnue.edu.vn OPTIMIZATION OF TOTAL SAPONIN EXTRACTION FROM TAM THAT ROOTS USING THE DESIRABLITY METHODOLOGY Nguyen Quang Tuyen and Dang Ngoc Quang Faculty of Chemistry, Hanoi National University of Education Abstract: In Vietnam, Tam that hoang (Panax stipuleanatus Tsai et Feng) belonging to Araliaceous family is a valuable medicinal plant that possesses many biologically active compounds. Among them, flavonoids, polysaccharides, and saponins are the major constituents of this plant. They have anti-oxidant, anti-microbial, anti-cancer, and tonic properties. In this paper, we studied the factors that affect the process of total saponin extraction from Tam that roots originated from Sapa town, Lao Cai province. The total saponin extraction from Tam that roots was optimized using the desirability methodology with the experimental order of Box-Behnken design. The concentrations of ethanol, the ratio of ethanol/material (v/m), the time of ultrasonic extraction with the frequency of 33 - 40 kHz are the three main considered factors. The optimized condition for total saponin extraction was found as follows: ethanol concentration 65%, ethanol/material ratio 55/1(v/m), and extraction time 75 minutes. In this condition, the maximum total saponin content was predicted to be 15.01%. Keywords: Extraction, saponin, Panax stipuleanatus.1. Introduction Extraction is the first step of great importance for the recovery and purification of activeingredients from plant materials. Traditional extraction techniques usually require many extractionhours and have low efficiency. Recently, there have been numerous reports on the application ofhigh intensity or power ultrasound in the extraction of various phyto-chemicals such as alkaloids,flavonoids, poly-saccharides, proteins and essential oils from various parts of plants and their seeds [1-3].The ultrasound-assisted extraction can be done at a low temperature to avoid the decomposition ofnatural compounds and essential oil. Even so there are several advanced extraction technologies,ultrasound-assisted extraction using ethanol solvent is a reasonable choice, since it is economicaland can be applied in industry without toxic residues [4-7]. Statistical experimental designs are proved to be more satisfactory and effective than othertraditional which only allow us to study one-at-a-time factor or mathematical methods because itcan be used to study many variables simultaneously, with less observation. This method has beenwidely used in different research fields such as biology, agriculture, food, and medicine etc.Received October 10, 2016. Accepted November 25, 2016.Contact Dang Ngoc Quang, e-mail address: quangdn@hnue.edu.vn26 Optimization of total saponin extraction from Tam that roots using the desirablity methodologyDesirability methodology is considered one of the effective methods for multiple objectiveoptimizations of technical factors [8]. In this study, the desirability methodology was employed tooptimize total saponin extraction from dried powder of the roots of Panax stipuleanatus usingethanol as solvent.2. Content2.1. Experiment2.1.1. Material Roots of Panax stipuleanatus, were collected in Sapa town, Lao Cai province, northern partof Vietnam in May 2013, with samples identified by Mrs. Pham Thanh Huyen from NationalInstitute of Medicinal Materials. Voucher specimens were deposited at the Faculty of Chemistry,Hanoi National University of Education (N0-9754; 9755). Materials were collected, cleanedsteamed at 100 0C for 5 minutes, and sliced, then dried at 60 0C for 5 days to the moisture of 5 - 6%,finally powdered with the size of particles of 0.1 - 1 mm. Dried samples were kept in two-layerpolyethylene bags, placed into glass bottles and stored in a dry and cool place. 2.1.2. Analysis methods Quantitative analysis of the total saponin was carried out using the analytical balance. Inaddition, the saponin content was determined by HPLC (Hitachi) with standard saponins(notoginsenosid R1, ginsenosid Rg1 and ginsenosid Rb1), equipped with a reversed phase column,phenomenex C18 (5 m × 250 mm × 4.6 mm) using acetonitrile and water as a solvent system.2.1.3. Research methods Extraction: Dried powder of Tam that roots (approximately 2 g) was placed in a 250 mLflask. The extraction was carried out in a thermo-regulatory ultrasound machine with an ethanolconcentration 50 - 90%, ethanol/material ratio (30 - 60/1, v/m), and extraction time 30 - 90minutes. The extract was filtered and solvents were removed by a rotary evaporator. The residuewas finally dried by a vacuum dryer. The total saponin was calculated by a percentage of thesample (% dry substance). Statistical experimental design method: Using the statistical Box-Behnken design with threefactors: ethanol concentration (%), ethanol/material ratio (v/m) and ultrasonic extraction times(minutes), (Table 1). This design has 15 trials including 3 ones for center points to response: totalsaponin (% dry substance), (Table 2). Table 1. The factors and their levels Levels Factors Symbols Units -1 0 +1 Ethanol A % 50 70 90 ...