Research’s objective
This research aims to identify the factors influencing the extraction process of Polycias Fruticosa root and leaf, with the goal of enhancing the efficiency of pure flavonoid extraction from these plant parts.
The main objectives of this thesis are briefly summarized in the followings:
- To determine total flavonoid content in Polycias Fruticosa
- Qualitative analysis of total flavonoid content found in Polyscias fruticosa root and leaf
This study aims to explore the key factors affecting the extraction of total flavonoids from the roots and leaves of Polyscias fruticosa Important variables include extraction temperature, solvent type, concentration, extraction duration, and material size, all of which may significantly influence the efficiency of flavonoid extraction.
Research question
- Do the roots and leaves contain total flavonoids?
- If having, what factors can influence the extraction process from Polyscias fruticosa root and leaf?
Limitation
- Due to the outbreak of Covid-19 pandemic, I had trouble travelling which made it harder to do experiments in the laboratory
- The raw material was harvested in the different time so there will be difference chemical concentration.
LITERATURE REVIEW
Overview about Polycias Fruticosa (L.) Harms
2.1.1 Characteristics of Polycias Fruticosa (L.) Harms Polyscias fruticosa is also known as Ming aralia, which is a member of the
The Aralioideae subfamily includes the Polyscias genus, which belongs to the Araliaceae family within the Plantae kingdom Polyscias fruticosa is a perennial evergreen shrub that typically reaches heights of 0.8 to 1.5 meters and features smooth spines Its dark green leaves are glossy and exhibit a tripinnate structure The plant produces flat fruits measuring 3-4mm in length and 1mm in thickness, each with a distinct spout.
Polyscias fruticosa is a popular plant grown as an ornamental throughout in
Vietnam is home to the Polyscias fruticosa tree, which also grows in Laos and southern China This perennial tree, which can live for several decades, is typically harvested in autumn when its roots, stems, leaves, and branches are collected from trees that are over three years old After harvesting, these parts are sliced and dried for future use Polyscias fruticosa thrives in humid environments and prefers temperatures below 28°C, making it a popular choice for planting in temples, family gardens, and hospitals.
Polyscias fruticosa, valued both for its ornamental appeal and medicinal properties, is increasingly sought after for home gardens, office bonsai, and interior decor This versatile plant not only enhances aesthetic environments but also offers significant health benefits, including improved circulation, pain relief from rheumatism, and various medicinal effects such as diuretic, anti-depressant, anti-inflammatory, antipyretic, and enzyme inhibitory properties.
Polyscias fruticosa, there are alkaloids, glucosides, saponins, flavonoids, tannins, vitamin B1, amino acids including lysine, xystei, and methionine which are irreplaceable amino acids
2.1.2 Overview of the chemical composition of Polycias fruticosa (L) Harms Polycias fruticosa contains alkaloids, glucosides, saponins, flavonoids, tannins, vitamin B1, and essential amino acids such as lysine, cysteine, and methionine The Polycias fruticosa root has the greatest chemical components, whereas the leaves, branches, and stems have smaller quantities (Do Tat Loi, 2004)
The root bark and leaves are rich in various beneficial compounds, including saponins, alkaloids, vitamins B1, B2, B6, and C, as well as 20 amino acids, glycosides, phytosterols, tannins, organic acids, essential oils, trace elements, and 21% sugar Additionally, the leaves are noted for containing 1.65% triterpene saponins, specifically oleanolic acid (Do Huy Bich et al, 2006).
The National Institute of Medicinal Materials' Research Center has identified five polyacetylene compounds in the leaves of Panax ginseng, including panaxynol and panoxydol, which are unique to this plant and absent in other Panax and Araliaceae species While five polyacetylene chemicals were also found in the roots, only three—panoxydol, panaxynol, and heptadeca-1,8(E)-diene-4,6-diyn-3,10-diol—were consistent with those in the leaves These compounds are recognized for their strong antibacterial and anti-cancer properties (Do Huy Bich et al., 2006).
2.1.3 Overview of the pharmacological composition of Polycias fruticosa (L) Harms
Contemporary medicine highlights several significant benefits of the plant, such as its role as a general tonic, its ability to improve appetite and promote better sleep, and its effectiveness in aiding weight gain Additionally, it boosts vitality, enhances work capacity, supports recovery, activates nerve cells, and improves memory (Tran Thi Kim Tuyen, 2017).
Polycias fruticosa has the following effects in Oriental Medicine: detoxification, diagnostic, typhoid, urinary tract, lung cooling, hemoptysis, dysentery, rheumatism, limb aches and pains (Tran Thi Kim Tuyen, 2017)
The root of Polycias fruticosa is renowned for its tonic properties, effectively strengthening the body and addressing issues such as weakness, poor digestion, and insufficient milk production in postpartum women Additionally, it serves as a remedy for coughs, uterine discomfort, and acts as a diuretic and anti-toxic agent The leaves are utilized to treat various ailments, including colds, fevers, swollen boils, allergic rashes, and wounds through peeling and rubbing In contrast, the stems and branches provide relief from rheumatism and back pain In India, this plant is recognized for its astringent and antimalarial effects, while its roots and leaves are employed to alleviate kidney and bladder stones, as well as dysuria For wound treatment, leaf powder mixed with salt is applied.
Overview of flavonoids
Flavonoids is a group of natural substances with variable phenolic structures, are found in fruits, vegetables, grains, bark, roots, stems, flowers, tea and wine (A N Panche, 2016) Flavonoids are phenolic chemicals having a C 6 -
The C3-C6 structure is a fundamental framework consisting of two benzene rings, A and B, connected by a three-carbon chain This structure represents a class of natural chemicals commonly found in medicinal plants (Bui Hong Hanh, 2013).
Flavonoids were discovered in 1938 by a Hungarian scientist named Dr Albert Szent-Gyorgyi, who referred to them as “vitamin P”
In the 1930s, Albert Szent-Gyorgyi and his team discovered that fresh extracts from citrus fruits like oranges and lemons were more effective than vitamin C alone in preventing scurvy They attributed this enhanced effect to other compounds in the extracts, naming them "citrin" or "Vitamin P" due to their role in reducing capillary permeability However, later research revealed that these compounds, including hesperidin and eriodictyol, did not meet the criteria for a vitamin, rendering the term obsolete (Bui Hong Hanh, 2013).
2.2.3 Structure and classification of flavonoids 2.2.3.1 Structure of flavonoids
Flavonoids are polyphenolic chains made up of 15 Carbon atoms and two benzene rings joined by a three-carbon line
The above structure may be represented as a C 6 -C 3 -C 6 system
Flavonoids have a chemical structure that is based on a 15 Carbon framework with a 2 nd , 3 rd , or 4 th B aromatic chromane
Figure 2.2 Structure of flavonoid aromatic ring Flavonoids are made up of two aromatic rings and one pyran ring:
Aromatic ring A is the aromatic ring on the left
Aromatic ring B is the aromatic ring on the right
A pyran ring is an intermediate ring that contains an oxygen atom (Bui Hong Hanh, 2013)
Flavonoids are categorized into various subgroups based on the structure of their C ring, particularly where the B ring attaches, as well as the levels of unsaturation and oxidation present in the C ring One notable subgroup is isoflavones, which are a specific type of flavonoid characterized by their unique structure.
The B ring is connected to the C ring at position 3, while connections at position 4 classify compounds as neoflavonoids Additionally, compounds with the B ring linked at position 2 can be categorized into various subgroups based on the structural features of the C ring, including flavones, flavonols, flavanones, flavanonols, flavanols (or catechins), anthocyanins, and chalcones (A N Panche, 2016).
Flavonoids are classified into three major groups based on the structure of the Carbon chain in the C 6 C 3 C 6 skeleton:
Flavan, flavan 3-ol (catechin), flavan 4-ol, flavan 3,4-diol, flavone, flavanol, flavonol, chalcon, anthocyanin, anthocyanidin, aurone are examples of eucaflavonoid (2-phenylbenzopyrans)
Isoflavan, isoflavan-4-ol, isoflavones, isoflavanone, rotenoid are all isoflavonoid (3-benzopyrans)
Calophylloid, neoflavan, and other neoflavonoid (4-benzopyrans) (Bui Hong Hanh, 2013)
Figure 2.3 Basic skeleton structure of flavonoids and their classes
2.2.4 Overview of properties of flavonoids 2.2.4.1 The physical properties of flavonoids
Physical characteristics play a crucial role in the isolation, analysis, and identification of flavonoid molecules Flavon derivatives and flavols typically exhibit light yellow to yellow hues, while chalcones and auros range from dark yellow to orange-red Other compounds such as isoflavones, flavanones, isoflavanols, flavanonols, leucoantocyanidins, and catechins are generally colorless In contrast, anthocyanidins display a spectrum of colors—yellow-orange, red, and purple—depending on the environmental pH (Nguyen Minh Thang, 2009).
Solvent solubility: Flavonoid' solubility varies based on the OH group and other substituents Flavonoids are found in plants mostly in two forms:
Flavonoid aglycol is a flavonoid free form The property of flavonoid aglycol is dissolved in organic solvents such as ether and ethanol Water does not dissolve flavonoid aglycol
Flavonoid glycosides are flavonoids that have been connected to sugar - glucid Flavonoid glycosides are dissolved in water Flavonoid glycosides are insoluble in non-polar organic solvents
Flavonoids possess a significant capacity for absorbing Ultraviolet (UV) radiation, attributed to their unique structure featuring a conjugated double bond system formed by two benzene rings (A and B) and a pyran (C) ring These compounds exhibit two primary absorption bands: the first band occurs at wavelengths exceeding 290 nm, while the second band is found within the 220 to 280 nm range (Nguyen Minh Thang, 2009).
In our research on flavonoid extraction, we selected ethanol as the solvent due to its rapid denaturing effect and ability to break down cell membranes, creating optimal conditions for flavonoid extraction and antioxidant exposure Additionally, ethanol's unique structure, featuring both a polar and a non-polar end, aligns well with the characteristics of flavonoids.
2.2.4.2 The chemical propeties of flavonoids
Flavonoids possess a complex chemical structure, leading to significant variations in their chemical reactivity influenced by factors such as the location of hydroxyl (OH) groups, the presence of conjugated double bond systems, and various substituents (Dias et al., 2021) The essential responses of flavonoids are outlined below.
The OH- group reacts in three ways: oxidation, hydrogen bond creation, and esterification
Diazotization is an aromatic ring reaction
The complexation of carbonyl groups with metals such as Fe, Zn, and Mg involves an oxidation-reduction reaction, resulting in products that exhibit distinct orange, pink, or red colors This reaction specifically applies to flavonoids containing a C=O group at position C4 and a double bond between C2 and C3 (Nguyen Minh Thang, 2009).
2.2.5 The biological values of flavonoid Antioxidant activity
Flavonoids are recognized for their antioxidant properties, which can be attributed to their molecular structure, particularly the number of hydroxyl substituents present; a higher number correlates with increased antioxidant activity (Mercia Marques Juca, 2018) These compounds have been integral to Eastern medicine for centuries, valued for their protective characteristics Common sources of flavonoids in Eastern medicine include scutellaria root, fennel berries, licorice, and green tea (Falcones Ferreyra et al, 2012).
Inflammation is a natural response to harm, but it requires careful management to prevent overactivity of the immune system Chemical flavonoids play a significant role in mitigating excessive inflammation, while various plant-derived substances, including polysaccharides, lectins, peptides, saponins, and oils, can stimulate the immune system and exhibit immuno-modulatory effects Research has explored the impact of flavonoids on various immune cells, including B and T lymphocytes, macrophages, natural killer (NK) cells, basophils, neutrophils, eosinophils, and monocytes (Mercia Marques Juca, 2018).
Flavonoids, a type of polyphenol found in plants, play a crucial role in antimicrobial defense, exhibiting antibacterial properties against various pathogens in vitro Additionally, they possess antioxidant capabilities and have demonstrated significant anticancer effects (Mercia Marques Juca, 2018).
Flavonoids exhibit antiviral activity primarily through their antioxidant properties, which include blocking enzymes, damaging viral cell membranes, limiting virus penetration, and enhancing host defense mechanisms Among the flavonoid class, compounds like catechins, quercetin, epicatechins, and theaflavins are particularly valued for their potent antiviral effects (Mercia Marques Juca, 2018).
Flavonoids possess antibacterial properties attributed to their hydroxyl phenolic groups, which bind to proteins and inhibit bacterial enzymes, disrupting their production pathways.
Biofilms play a crucial role in antibacterial action, as they are produced by pathogenic bacteria and contribute to various health issues These structures are essential in bacterial pathogenesis and antibiotic resistance, making biofilm inhibitors important for controlling infectious diseases Research has shown that flavonoids can effectively inhibit biofilm formation in bacteria such as Streptococcus mutans, Aeromonas hydrophila, and Escherichia coli Specific flavonoids, including naringenin, kaempferol, and quercetin, have demonstrated the ability to suppress biofilm development in E coli Additionally, a study by Lee et al (2011) found that afloretin, a natural flavonoid, serves as a non-toxic inhibitor of E coli biofilms.
Overview of methods for the total flavonoid extraction
The flavonoid extraction process utilizes principles similar to those of polyphenol extraction, typically employing solvents such as methanol, ethanol, acetonitrile, acetone, or their combinations with water The choice of solvent polarity is crucial and varies based on the specific types of flavonoids being extracted (Milena Tzanova et al).
For less polar flavonoids like isoflavones, flavonones, and flavones, suitable solvents include acetone, chloroform, methylene chloride, and diethyl ether In contrast, more polar flavonoid fractions typically require alcohol or a combination of alcohol and water as solvents.
The extraction process is carried out by different methods including:
The traditional extraction methods: Soxhlet extraction apparatus, thorough extraction, and progressive infiltration
The mordern extraction methods: Extraction with supports from
2.3.1 The traditional extraction methods 2.3.1.1 The Soxhlet extraction apparatus
Soxhlet extraction is a widely utilized technique for extracting analytes from solid materials, and its conventional procedure has been a staple in analytical laboratories since its inception.
1879 To this day, the Soxhlet extraction technique is still used to compare the effectiveness of current extraction techniques
Soxhlet extraction is a high-temperature continuous extraction method that utilizes a Soxhlet apparatus In this process, a soil sample is placed in a porous measuring tube, while the extraction solvent is heated in a bottom flask The solvent evaporates, passes through the sample tube, condenses in a condenser, and then drips back, making this method efficient in terms of solvent use and processing time However, there are notable drawbacks: the flavonoid extraction must be heat stable, plant samples need to be dried, and the method requires the use of toxic and flammable organic solvents.
Medicinal herbs are soaked in a solvent, allowing the extraction process to occur over a specified duration based on the type of herb During this process, the extract drips to the bottom while fresh solvent is continuously added from the top, ensuring a slow and steady flow without stirring Typically, the solvent layer in the extraction vessel is maintained about 3-4 cm above the surface of the medicinal material.
Simple thorough method: A thorough method in which the active components in medicinal plants are extracted using a fresh solvent until they are exhausted
Fractional thorough method (re-thorough): This is a method approach that uses dilute fresh batches (new medicinal herbs) or extract batches with varying degrees of extraction
Conserve solvents (re-exhausted) Disadvantage of thorough method:
There are general drawbacks of fractional thorough method, including low productivity and physical labor
The process is more difficult than the immersion method
Consumption of solvents (simple thorough method)
During the extraction process, ground plant material is placed in a sealed container with an appropriate solvent The samples are maintained at room temperature for a minimum of three days, with frequent shaking to enhance the extraction This method allows the solvents to soften and break down the plant's cell walls, effectively releasing soluble phytochemicals.
Advantage of the maceration extraction method: The simplest technique, requiring no specific laboratory equipment
Disadvantage of the maceration extraction method: Massive solvent volume, lengthy processing time, and future purification required When it comes to purity, superior extraction technologies should be considered
There are four common types of modern extraction methods (Celeste De Monte et al, 2014)
Ultrasonic waves, with frequencies between 20 kHz and 10 MHz, can penetrate solids, liquids, and gases undetected by humans In the extraction process, high-energy gas bubbles are generated, disrupting cell wall structures and enhancing the release of intracellular materials To extract phenolic compounds from plants, ultrasonic waves are utilized through transducers and ultrasonic baths Key parameters such as amplitude, frequency, and wavelength, along with the devices' power and intensity, significantly influence the extraction efficiency Additionally, the design of the ultrasonic bath and the transducer shape play crucial roles in optimizing the extraction process.
Advantage of Ultrasonication Assisted Extraction (UAE)
Ease of use, low cost, high efficiency, low organic solvent consumption and shorter extraction time
On a large scale and industrial level, it may be employed as a simple and dependable process in a wide range of organic solvents for diverse phenolic compounds
Disadvantage of Ultrasonication Assisted Extraction (UAE): It must be done on a huge scale
Microwaves are electromagnetic waves with frequencies ranging from 30 to
At 300 MHz, electromagnetic induction generates a heating effect by continuously moving polar molecules within matter This movement leads to the breakdown of cell walls, resulting in the release of active chemicals into the surrounding environment.
Advantage of Microwave – assisted extraction (MAE)
Reduced time and expense, great efficiency, and little usage of organic solvents
Extract many compounds concurrently in a short amount of time
Disadvantage of Microwave – assisted extraction (MAE):
Microwave radiation can be reactive and are difficult to implement on a wide scale
In this process, the solvent operates above its critical temperature and pressure, eliminating surface tension and exhibiting both liquid and gas characteristics This unique state enhances its efficiency in extracting phenolic chemicals from plants The low viscosity and high diffusivity of supercritical fluids enable rapid and effective extraction of a wide range of phenolic compounds.
Advantage of Supercritical fluid extraction (SFE)
It is speedy, simple to operate, and selective
Heat-labile chemicals are permanent in this process, that needs little or no solvent
Disadvantage of supercritical fluid extraction (SFE): Expensive and only appropriate for high-value materials
PLE uses high pressure to keep solvents liquid above their boiling point
As a result, hydrophobic molecules in the solvent have a high solubility and diffusion rate, and the solvent penetrates deeply into the substrate
- Reduced extraction time and solvent consumption, as well as improved repeatability
- Automation, which allows for faster extraction with less solvent
Disadvantages: Must have appropriate equipment, which is more expensive
Factors influencing flavonoid extraction process
To achieve optimal extraction of desired compounds from various materials, it is essential to select the appropriate solvents The choice between organic or inorganic, as well as polar or non-polar solvents, depends on the specific substance being extracted Careful selection of the solvent is crucial for maximizing the concentration of the extract.
The extraction of materials relies on specific solvents, with their concentration significantly influencing the quantity of desired substances obtained Both high and low solvent concentrations can lead to either insufficient extraction of target compounds or the separation of undesirable chemicals Therefore, the selection of a solvent is crucial, taking into account its solubility, polarity, and concentration levels.
Extraction time plays a crucial role in the extraction process, influencing the quantity of material extracted It is essential for the extraction time to be sufficiently long to ensure effective separation of compounds from the substance.
As a result, the extraction time is chosen based on the nature of the extraction and the kind of solvent
The solvent-to-material ratio significantly impacts extraction efficiency by enhancing the diffusion of soluble components into the solvent Increasing this ratio boosts the concentration difference, facilitating the extraction process However, excessively high solvent volumes can complicate post-extraction processing Therefore, selecting the optimal solvent ratio is crucial for maximizing extraction efficiency, tailored to the specific substances targeted from the raw materials.
Crushing the material disrupts its tissue structure, facilitating chemical extraction The particle size affects the contact area with the solvent, impacting extraction efficiency; smaller particles enhance efficiency but can lead to blockages in capillary tubes due to chemical deposits, resulting in contamination and complications in subsequent processes.
Research situation in the world and Vietnam
In 1998, Bensita Mary Bernard et al discovered that n-Butanol extract from
Polycias fruticosa leaves had anti-inflammatory properties and can decrease edema in mice
M.B Bensita et al discovered the antibacterial activity of polyacetylene chemicals in Polycias fruticosa leaves in a paper published in the journal Ancient Science of Life in 1999 This antibacterial ability outperforms saponins
George Asumeng Koffuor et al wrote in 2016 on the potential of ethanol extracts from Polycias fruticosa leaves to cure asthma
Vo Xuan Minh investigated and suggested the technology of extracting saponins from Polycias fruticosa and generating a number of dosage-form products from this substance in 1992
Nguyen Thi Ngoc Thuy et al (2020) explored the extraction of total triterpenoid saponins from Polycias fruticosa leaves using cellulose enzymes, finding that enzyme-treated samples outperformed untreated ones The study identified the material-to-solvent ratio as a key factor for effective extraction and the ability to inhibit amylase Optimal conditions included a water-to-enzyme ratio of 1:30, an enzyme-to-substance ratio of 1%, and a processing time of 40 minutes The use of ultrasonic methods significantly enhanced the extraction of beneficial components from Polycias fruticosa.
MATERIALS, RESEARCH CONTENTS AND METHODOLOGY 18 3.1 Material and research scope
Material
province after 3 years of cultivation The fresh root and leaf samples are cleaned and removed from broken slides After that these materials have been drying for
8 hours at 60 o C until the moisture content reaches under 10% The root and leaf were preserved in PE bags in the refrigerator and used during the period of this research.
Research scope
Research was carried out in the laboratory scale.
Workplace and time to proceed
Location: Laboratory under the Institute of Life Sciences, Thai Nguyen University
Implementation time: February 2022 to October 2022
Chemicals, equipment
Research content
This article presents a qualitative analysis of the total flavonoid content in the roots and leaves of Polyscias fruticosa It also explores the various factors influencing the extraction process of flavonoids from these plant parts, providing insights into optimal extraction methods for maximizing flavonoid yield.
To choose the most appropriate solvents for extraction process of total flavonoid content in the Polyscias fruticosa root and leaf
To determine of concentration of solvent on total flavonoid content extraction process
To investigate on the effect of solvent/material ratio on the total flavonoid content extraction process
To investigate on the effect of extraction time on the total flavonoid content extraction process
To investigate on the influence of raw material size on the total flavonoid content extraction process
To investigate on the effect of magnetic stirrer on the total flavonoid content extraction process.
Research methods
3.5.1 Experimental design method Experiment 1: Qualitative analysis of total flavonoids found in the Polyscias fruticosa root and leaf
To extract flavonoids from Polycias fruticosa, grind the root and leaf, then soak the powder in 70% ethanol for 20 hours at room temperature After filtering, collect 1ml of the extract and add 1ml of 10% Pb(CH3COO)2 solution Allow the mixture to stand for 1-2 minutes to observe any reactions The presence of a yellow precipitate indicates that flavonoid components are present in the extract.
Experiment 2: To investigate the most appropriate solvent for total flavonoid extraction process
To investigate the influence of extraction solvent type, two different solvents are used: water (H 2 O) and ethanol (C 2 H 5 OH) The experiments are set up according to the following table:
Table 3.3 The investigation of the appropriate solvents for total flavonoid extraction
Formula Experimental criteria Extraction conditions
1 Extraction of 0.1g of leaf ground powder and 0.5g of root ground powder with distilled water (H 2 O) and ethanol (C 2 H 5 OH)
Ratio: 100:1 (ml/g) for leaf and 50:1 (ml/g) for root Time: 20 hours
At the end of the process, determine the total flavonoid content, and select the optimal solvent The results of experiment 2 are used for the next experiment
Experiment 3: To investigate the influence of solvent concentration on total flavonoid extraction process
To investigate the influence of extraction solvent concentration on the flavonoid content at 70%, 80%, and 90% concentrations The experiments are set up according to the following table:
Table 3.4 The investigation of the influence of solvent concentration on total flavonoid extraction
Formula Experimental criteria Extraction conditions
1 0.1g of leaf ground powder and 0.5g of root ground powder were soaked with solvent determined at different concentration of 70%, 80% and 90%
Ratio: 100:1 (ml/g) for leaf and 50:1 (ml/g) for root Time: 20 hours
The solvent is determined in experiment 2
At the end of the process, determine the total flavonoid content, select the optimal solvent concentration The results of experiment 3 are used for the next experiment
Experiment 4: To investigate the effect of solvent time on total flavonoid extraction process
To investigate the influence of extraction solvent time on the flavonoid content at for 16; 18 20; 22 and 24 hours The experiments are set up according to the following table:
Table 3.5 The investigation of the influence of solvent time on total flavonoid extraction
Formula Experimental criteria Extraction conditions
1 0.1g of leaf ground powder or
0.5g of root ground powder samples were soaked with solvent for 16, 18, 20, 22, and
Ratio: 100:1 (ml/g) for leaf and 50:1 (ml/g) for root Time: 20 hours
The solvent is determined in experiment 2
The solvent concentration is determined in experiment 3 Temperature: room temperature
At the end of the process, determine the total flavonoid content, select the optimal extraction time The results of experiment 4 are used for the next experiment
Experiment 5: To investigate the influence of solvent/material ratio on total flavonoid extraction process
To investigate the influence of solvent/material ratio on the flavonoid content at 5:1, 10:1, 30:1, 50:1, 70:1 and 100:1 The experiments are set up according to the following table
Table 3.6 The investigation of the influence of solvent/material ratio on total flavonoid extraction process
Formula Experimental criteria Extraction conditions
1 Extraction of 0.1g of leaf and
0.5g of root ground powder with solvent/material ratio determined at 5:1 (ml/g), 10:1 (ml/g), 30:1 (ml/g), 50:1 (ml/g), 70:1 (ml/g) and 100:1 (ml/g)
The solvent is determined in experiment 2
The solvent concentration is determined in experiment 3 The extraction time is determined in experiment 4 Temperature: room temperature
At the end of the process, determine the total flavonoid content, select the optimal solvent/material ratio The results of experiment 5 are used for the next experiment
Experiment 6: To investigate the influence of raw material size on total flavonoid extraction process
To investigate the influence of raw material size with size 1mm, 1 < size 3mm, 3