Solid Liquid Extraction of Ganoderma Lucidum Essay

M. Dinithi Ishara Wijetunge 1. 0 Summary The solid-liquid extraction of Ganoderma Lucidum is carried out to in order understand the different techniques utilized. Furthermore, it is used to understand the effects of various physical factors on the level of extraction. An intense literature research was carried out on the specimen itself to understand its sources, bioactive constituents, growth and cultivation and medical properties. The basic meaning of the term solid-liquid extraction (SLE) is identified and the basic factors affecting is rate are outlined to be particle size, solvent, temperature and agitation.

The methods of SLE utilized in the experimental procedure are (i) conventional- hot plate magnetic stirrer method and non conventional (ii) ultra-sound bath and (iii) ultra sound probe method. The underlying theory of the conventional method is that SLE is favored for increasing temperatures and longer agitation periods. Moreover, for the non-conventional methods, the mechanism of ultrasound radiation in a liquid medium is understood, outlining clearly the meaning and uses of micro -bubble cavitation. The mechanism by which the SLE extraction occurs when subjected to ultrasoun d radiation is found.

The advantages of using ultra-sound assisted extraction equipment as opposed to conventional SLE methods are identified. Although non-conventional ultra-sound equipment is more effective, they are expensive thus their applicability for SLE is reduced. For each piece of equipment, the working mechanism is identified in relation to how it aids in the SLE of the G. Lucidum specimen. Finally the method of analysis is reviewed, where the working mechanism of the UV/VIS spectrometer is outlined. It is shown how the device utilizes the sample and reference cells to generate graphs of absorbance vs. avelength. In the experimental section, the preparation procedure of the process mixture of 0. 5g G. lucidum/100 ml distilled water is explained. Thereafter, the experimental procedure for using each equipment is outlined. The experimental difficulties faced were mentioned such as delay in recording results, disturbance of magnetic agitator (magnetic hot plate stirrer) when withdrawing samples etc. For each equipment a simple schematic diagram explaining its integral parts are included together with pictures of the actual equipment used during the laboratory session. 4|Page

We Will Write a Custom Essay Specifically
For You For Only $13.90/page!


order now

H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge 2. 0 Introduction This report is based in the solid liquid extraction of the mu shroom Ganoderma Lucidum. The solid liquid extraction of the plant tissue is carried out experimentally using conventional hot plat e magnetic stirrer and non-conventional methods using an ultrasonic bath and horn apparatus available in the laboratory. For each equipment used, specific process variables are varied in order to test and observe their effect on the solid -liquid extraction.

The overall objective of the experiments performed is: analyzing the extraction of bioactive constituents from the G. lucidum specimen using distilled water as the extractive solvent utilizing both conventional and non conventional methods. An individual objective was set for each equipment so as to test the affect of relevant parameters on the extent of SLE. For the magnetic hot plate stirrer, the objective was to test the effect of stirrer speed and time on the extent of SLE while other parameters such as concentration were kept constant. For the ultrasound bath the effect of 3 parameters were tested on the level of SLE. . e. the frequency, power and time while the temperature and concentration were maintained constant. Finally for the ultrasonic probe horn, the effect of amplitude and time were tested for their effects of SLE w hile the concentration was maintained constant. 5|Page H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge 3. 0 Literature Search 3. 1 Sources of Ganoderma Lucidum “Ganoderma Lucidum is a Woody basidiomycotina mushroom belonging to the family of Ganodermaceae of polyporales. (Deepalakshmi and Mirunali 2011) It is a form of annular mushroom. Its growth is common in Europe, Asia and North and the temperate region of South America. There are over 250 species of Ganoderma worldwide of which the species used in therapeutic applications is Ganoderma Lucidum. (Deepalakshmi and Mirunali 2011). It is considered as a herb which could be treated against all diseases and furthermore improve one’s longevity. (Wei Xu et al. 2009) Figure 3. 1. 1: Fruiting Body of Ganoderma Lucidum (Deral -Phytochi 2011) The following mushroom can be obtained by two methods: i) Natural resources: which grows on a large array of dead or dying trees. , eg. , Deciduous trees (e. g. oak, maple, elm, willow, sweet gum, magnolia and locust) and less commonly found on coniferous tree (e. g. , Larix, ptea, pinus). (Deepalakshmi and Mirunali 2011) Its growth is common in Europe, Asia and North and the temperate region of South America. However due to its increasing demand, G. lucidum is scarce in nature. (Boh at al. 2007) (ii) Artificial resources: Due to its scarcity in nature, most companies have to resort to artificial cultivation.

The primary methods of Ganoderma cultivation is: ? Saw dust cultivation in bags and bottles – Since G. lucidum is a lignin-degrading whiterot fungus which grows on hardwoods and woody tissue, such as sawdust (Boh at al. 2007) ? On natural logs – This involves both long unsterilized logs used in the past and short sterilized logs popularized since the 1980’s. However short log cultivation is preferred since it gives a higher yield in a shorter cultivation time enabling quicker turnover of the capital. (Boh at al. 2007) Both cultivation techniques depend on the same environmental factors, ncluding temperature, humidity and oxygen (Boh at al. 2007) . A more recent development of G. Lucidum cultivation is 6|Page H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge the cultivation of its mycelia in bioreactors on either solid substrates and in liquid submerged culture” (Xu et al. 2007) 3. 2 Bioactive constituents of Ganoderma Lucidum The bioactive constituent of Ganoderma lucidum originates from its chemical composition. The Chemical constituents present include: (i)

Polysaccharides –water-soluble branched arabinoxyloglucan, water-soluble heteroglycan, several water-insoluble heteroglucans and ganoderans; (Khan and Abourshed 2010) (ii) Polysaccharide- peptide complex : lectins and glycopeptides with carbohydrate-topeptide ratio of approximately 10:1; (Khan and Abourshed 2010) (iii) Triterpenoids (mainly lanostane type) including ganoderic acids, lucidenic acids, lucidones, lucialdehydes, ganolucidic acids, lucidumol, ganoderal, ganoderiols, ganoderols, ganodermanontriol and ganodermatriol (Khan and Abourshed 2010) (iv) Proteins: water-soluble polypeptides and amino acids; (Khan and Abourshed 2010) (v)

Trehalose and other sugars including mannitol; betaine; denosine; (Khan and Abourshed 2010) (vi) Sterols: mainly ergosterol and ergosterol peroxide, b-sitosterol, 24-methylcholesta7,22-dien-3-b-ol, and other sterol esters like fungal lysozyme, acid protease ; (Khan and Abourshed 2010) (vii) Enzymes such as laccase, endopolygalacturonase, cellulase, amylase, etc. (Khan and Abourshed 2010) (viii) Alkanes including tetracosane and hentriacontane;(Khan and Abourshed 2010) (ix) Fatty acids including tetracosanoic, stearic, palmitic, onadecanoic , and behenic acids (Khan and Abourshed 2010) x) Major inorganic elements present include Ca, Mg, Na, Mn, Fe, Zn, Cu, and Ge (Khan and Abourshed 2010) (xi) Essential oil hydro distillate. (Khan and Abourshed 2010) It is important to note the two most important bioactive constituents with the most pharmacological uses are polysaccharides and triterpenoids. (Boh at al. 2007) The nutritional composition of a G. lucidum is affected by “differences among strains, composition of growth substrate, method of cultivation, stage of harvesting, specific portion of the fruiting bodies used for analysis. (Deepalakshmi and Mirunali 2011). 3. 3 Pharmacological Properties The pharmaceutical effects of triterpenoids are relief against hepatoprotective, hypertensive, hypocholesterolemic and histaminic effects. Also it improves immunity against tumors and other cancers (Lindequist et al. 2005), engiogenic activities and platelet aggregration in the body. In the case of polysaccharides, it too improves one’s immunity against tumors. “In addition, polysaccharides have a protective effect against free radicals and reduce cell damage caused by mutagens. (Khan and Abourshed 2010) 7|Page H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge 3. 4 What is solid- liquid extraction (SLE)? It is “the extraction of a soluble constituent from a solid by me ans of a solvent. ”(Richardson et al. 2002) Another commonly used term for SLE is “leaching”. The process consists of 3 integral parts: (i) Diffusion of the solvent through the pores of the solid (ii) The diffused solvent dissolves the solutes (i. e. transfer the solute to the liquid phase). iii) Transfer of the solution from porous solid to the main bulk of the solution. The slowest of the above three steps may act as the limiting step for the extraction rate . However, the first process has a negligible effect on the overall rate since it occurs so rapidly. .”(Richardson et al. 2002) 3. 3 Factor affecting the rate of SLE In general, there are 4 rate determining factors in SLE: (i) Particle size: As the particle size decreases, the interfacial surface area between the solid and liquid increases. As a result, rate of mass transfer during the extraction increase.

Also, the distance that the solute must diffuse within the solid decreases. (Richardson et al. 2002)In the case of the experiment the G. Lucidum specimen used is in a ground up form (similar to saw –dust). (ii) Solvent: The liquid should have a low viscosity allowing it to flow freely through the particle medium. As the extraction proceeds, more and more solute will diffuse into the solvent thus reducing the rate of extraction with time. This is because the concentration gradient drops and also the liquid becomes more viscous. Richardson et al. 2002) (iii)Temperature: An increase in temperature increases the solubility of the solute being extracted, thus increasing the rate of extraction. Furthermore, in increase in temperatur e increases the diffusion coefficient of the solute within the solvent. (Richardson et al. 2002) (iv)Agitation of the fluid: “Agitation of the solvent is important because this increases the eddy diffusion and therefore the transfer of material from the surface of the particles to the bulk of the solution. ” (Richardson et al. 002) Also, the agitation process reduces the sedimentation of fine particles thus allowing a more effective use of the interfacial surface. 3. 4 Methods of SLE For the following experiment, the method of extraction used can be classified according to: (1) Conventional methods: – using a hot-plate stirrer apparatus. (2) Non- conventional extraction techniques: – using an ultra sound bath apparatus – Using an ultrasonic horn equipment 3. 4. 1 Conventional method: hot plate stirrer (magnetic stirrer) “Hot-water extraction is the most widely used technology for polysaccharide extraction ” (Quan Huang et al. 2010).

However, it is not the most efficient technique of SLE as it results in lower yields compared to the non-conventional methods mentioned above. 8|Page H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge The hot plate magnetic stirrer is used to heat up liquids and stir them with a magnetic stirring knob. This agitation of the solid-liquid mixture is the factor that assists in the extraction of components from the G. lucidum particles. The equipment consists of two knobs; one that controls the stirrer speed and another which controls the temperature.

The ma gnetic stirrer is used agitate non-viscous solutions with the aid of magnetic stirrer. The hot plate stirrer can be used in a temperature range between 40-200 oC. (Gelosa and Sliepcevich 2000) However, the extraction of bioconstituents from plant tissue using hot-water extraction results in “lower yields, longer extraction times and high temperatures. ” (Quan Huang et al. 2010) Therefore according to most literature, the solution for this is to resort to more novel extraction technologies such as ultra sound assisted extraction.

This sort of non- conventional extraction technique can accelerate the extraction of the bioactive constituents, while giving higher yields in a shorter period of time. (Quan Huang et al. 2010) 3. 4. 2 Non-conventional method: Ultrasonic extraction Ultrasound is a form of sound wave, and like all sound waves it requires a medium to travel. When sound waves travel, they create expansion and compression cycles in the medium (Garcia and de Castro 2003). During expansion, the ultrasound wave produces a negative pressure within the liquid.

If the ultrasonic wave is strong enough, this expansion results in micro bubbles. When these vapour bubbles form, grow and undergo implosive collapse, the phenomenon is known as “cavitation”. Cavitation is a nucleation process, which occurs at “pre-existing weak points of the liquid” (Garcia and de Castro 2003) e. g. in gas filled crevices in the suspended particulate matter within the liquid. At a certain point, the bubbles can no longer absorb any energy from the ultrasonic waves, thus they collapse. Figure 3. 4. 2. : Change in bubble size during the cavitation process until bubble collapse (Suslick 1994) This results in a rapid adiabatic compression process which produces very high temperatures. In regions where these bubbles collapse it creates what is known as a “sonochemical hot spot” (Suslick 1994). A hot spot of this nature has a temperature of approximately 5000 oC (similar to the surface temperature of the sun) and a pressure of1000 atm (similar to the pressure at the bottom of the ocean) with a lifetime less than a microsecond. Furthermore these hotspots have heating |Page H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge and cooling rates above 10 billion° C per second. (Suslick 1994) The temperature produced at these cavitation regions does not however affect the bulk of the liquid because the bubbles are very small in comparison and the heat from the implosion dissipate relatively quickly. (Suslick 1994) Figure 3. 4. 2. 2: The process of cavitation, from bubble formation till collapse (Suslick 1994) 3. 5 Theory of Ultra sound assisted Extraction (UAE)

Another commonly used term for UAE is leaching. When a solid liquid mixture is subjected to an ultrasonic source, after a certain period of time cavitation occurs within the liquid. “When cavitation occurs in a liquid close to the solid surface, the dynamics of cavity collapse change dramatically. ” (Garcia and de Castro 2003) In pure liquids, since the surrounding is uniform, the cavitation bubbles are spherical in shape. However when cavitation occurs adjacent to a solid surface, the collapse of the cavity is “asymmetric” producing jets of liquids at very high speeds.

The potential energy within the bubble is converted to kinetic energy causing the liquid jet of approximately 400 km/h to penetrate across the bubble. (Suslick1994) The effects of this kinetic activity in UAE is helps in the intensification of mass transfer within the solid liquid mixture, improvement of cell disruption, enhanced penetration and more efficient capillary effects(Vinatoru et al. 1997; Ozcan 2006) 3. 6 SLE extraction of G. lucidum using UAE Plant tissue such as G. lucidum is surrounded by cell wall s. The extraction process from dried material such as the G. ucidum involves 2 stages: (i) swelling and hydration of the plant tissue by the solvent (in this case water) and (ii) mass transfer of soluble constituents from the material into the solvent by diffusion and osmosis when the cell wall breaks (also known as rinsing). Both phenomena are favored by ultrasonic radiation. Furthermore, ultrasound aids in the enlargement of the pores of the cell wall thus improving the mss transfer. Ultrasound increases the swelling index of a material during the sonication process. The extractive value is much higher under ultrasound sonication rather than mechanical stirring.

A reduction in the particle size of the specimen increases the number of cells directly exposed to the ultrasonic cavitation. (Vinatoru 2001) For the G. Lucidum, the cavitation process, allows the extraction of mainly its carbohydrates and triterpenes together with other useful components mentioned earlier . (Vinatoru, 2001; Li et al. 2003) 10 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Figure 3. 6. 1: Schematic of the mass transfer process in plant cells during solid-liquid extraction (Vinatoru 2001) 3. 7 Advantages of Ultrasound assisted Extraction

In a journal article regarding ultrasound assisted extraction, Soni et al. (2001) identifies the advantages of UAE applications in solid-liquid extraction. 1) Ultrasound-assisted extraction is more effective and efficient in comparison to conventional extraction techniques. 2) Using ultrasound for solid-liquid extraction results in an increase of extraction yield and faster kinetics. 3) Ultrasound can also reduce the operating temperature allowing the extraction of thermolabile compounds which are those substances which is subject to destruction, decomposition or change in response to heat effects. ) Compared to other non- conventional leaching techniques such as microwave-assisted extraction, the ultrasound apparatus is cheap er and its operation is easier. 3. 8 Devices used for Ultrasonic Assisted Extraction There are two common devices for ultrasound application that is, the ultrasonic bath and the ultrasonic probe units. Figure 3. 8. 1: (From L to R) Side view of cavitation caused by ultrasonic horn (Industsonomechanics 2010), Top view of cavitation effects in water inside an ultrasonic bath (Sonixiv2012) 11 | P a g e H83 CEL Chemical Engineering Laboratory

Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge The more commonly used equipment is the ultrasonic bath due to its affordability. However, the use of this apparatus has 2 main disadvantages which can affect the reproducibility of the experimental results. They are: (i) lack of uniformity in the distribution of the ultrasound energy of since only small fraction of the liquid volume is in the immediate vicinity of the ultrasound source and (ii) it is not energy efficient since there is a decline of power with ti me.

In comparison, although ultrasonic probes are more expensive, they are more advantageous for use than the bath apparatus. That is, (i) they produce more effective cavitation since its energy is concentrated on a localized sample point. (Ozcan 2006) Table 3. 8. 1: Comparison of the ultrasonic probe and ultrasonic bath (Jianyong et al. 2001; Soni et al. 2001) 3. 9 Method of sample analysis; UV spectrometry In a conventional UV spectrometer, two samples are placed. One pure solvent (usually distilled water) sample which acts as the reference and another which is the sample of the process fluid to be analyzed.

The samples are held in a small square-section cell. The sample is scanned for approximately 30 s. During this period, radiation across the whole of the ultraviolet/visible range is scanned and radiation of the same frequency and intensity is simultaneously passed through a reference cell containing onl y the reference fluid. Then, photocells in the device detect the radiation transmitted and the spectrometer records the absorption by comparing the difference between the intensity of the radiation passing through the sample and the reference cells. (Faust, 1992) Figure 3. 9. How an ultraviolet spectrometer works (Faust, 1992) 12 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge 3. 10 Theory of UV/VIS spectrometry: Absorption laws To explain the absorption of molecules using a UV/VIS spectrometer, the “Beer -Lambert law” is used. It is a combination of the Beer’s law which shows that the absorption is proportional to the to the concentration of absorbing molecules and Lambert’s law which shows that the fraction of radiation absorbed is independent of the intensity of the radiation. Clark 2007). The Beer-Lambert law is represented by the following equation: (Faust 1992) (Faust 1992) Where: I0 – intensity of the reference cell I – intensity of the light passing through the sample cell. – molar absorption constant (mol-1dm3cm-1) – path length of absorbing solution (cm) – concentration of absorbing species (moldm-3) 13 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge 4. 0 Experimental Objectives of the experiment: ? ?

Extraction of bioactive constituents from Ganoderma Lucidum using distilled water as the extractive solvent using conventional and non-conventional extraction methods To study the effects of temperature, mixing speed, extraction time, ultrasonic power and frequency on the extent of solid-liquid extraction. Solid-Liquid extraction procedures: (1) Conventional method – Hot plate magnetic stirrer (2) Non- conventional method – Ultra sound bath apparatus – Ultrasonic horn equipment Preparation procedure ? ? ? ? ? ? ? Weigh 0. 5 g (error ±0. 002) of ground G. ucidum specimen using the electrical mass balance Measure 100 ml of pure distilled water into to a beaker Add the G. lucidum specimen to the beaker of 100 ml distilled water Stir using mixing rod for approximately 30 s. Cover the resulting mixture with paraffin paper (to avoid any liquid loss due to evaporation during the extraction process. ) Make a small hole in the middle of the paraffin sheet to allow a pipette to be added in order to withdraw liquid samples. The final sample: 0. 5 g Ganoderma Lucidum sample in 100 ml of the solvent water. 0. 5 g G. ucidum sample Figure 4. 1: Measuring a sample of 0. 5g in the electronic pan balance in the laboratory (±0. 001 error) 14 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Equipment 1. Conventional SLE extraction using hot plate magnetic stirrer The equipment consists of 2 knobs; one that controls the heat level (or temperature) and a second which controls the agitation. A magnetic stirrer was immersed in the solution of solid liquid mixture of G. lucidum and water.

When the beaker containing the SLE mixture is placed on the platform, the agitation action caused by the magnetic fields aids in the extraction of bioactive constituents from the mushroom tissue. Factor varied during the experiment are: – Speed of mixing (6 rpm and 12 rpm) – Time (15 minutes and 30 minutes) The criteria that were controlled at a constant value are: – the concentration (0. 5 g G. lucidum/ 100 ml water) Figure 4. 2: Simple schematic diagram of a hot plate stirrer (magnetic stirrer) (Mitchell 2008) Figure 4. 2: Actual image of the Hot plate stirrer (brand name: Cimarec) taken during laboratory session 5 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Experimental procedure: ? ? ? Perform the experiment on the hot plate stirrer at 2 different speeds; (i) at 6 rpm and (ii) at 12 rpm to test the effect of increasing agitation speed on the rate of solid-liquid extraction For each speed, the experiment is run for 15 minutes with sample extraction at 3 minute intervals and again for 30 minutes with sample extraction at 6 minute intervals. (Two experiment run-times were used in order to test the effect of time on the rate extraction. For the entire experiment using ultrasonic water bath, a total of 24 samples were collected. Experimental difficulties: When extracting the liquid sample for analysis in the UV spectrometer, the agitation pr ocess is affected. This is because when the pipette is inserted to the solid liquid mixture, it obstructs the movement of the magnetic stirrer. This reduces the reliability and accuracy of the results as the agitation won’t be uniform. A method of reducing this effect is to immerse the pipette in such a way that it does not knock against the stirrer during extraction of samples. 2.

Non conventional SLE extraction using Ultrasound Bath An ultrasound bath consists of 3 main components: (i) A tank – Which holds the water bath. The ultrasonic waves travel through the liquid medium held in the tank. The solid-liquid mixture in a beaker which needs to be subjected to the ultrasonic waves is immersed in the water bath with the aid of a clamp support. (ii) A power generator – power generator converts a standard electrical frequency (5 -60 Hz) into alternating frequency (over 20 kHz) (Ozcan 2006) (iii) An ultrasonic transducer – is a component which is bonded to the base of the ultrasonic bath.

It is of “magnetostrictive type or piezoelectric type” (Ozcan 2006). The task of the transducer in this case is to convert the electrical frequency to ultrasonic waves. These ultrasonic waves are then transferred to the water in the tank. For the ultrasonic bath, the criteria that were being varied are: -the frequency (35 kHz or 130 kHz) -the power (50% or 100%) – the time (15 minutes and 30 minutes) The criteria that were controlled at a constant value are: -the concentration (0. 5 g G. lucidum/ 100 ml water) -the temperature (40oC) 16 | P a g e H83 CEL Chemical Engineering Laboratory

Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Figure 4. 3: Schematic of ultrasound bath (Fuchs 1999; Ozcan 2006) 0. 5 g G. lucidum/100 ml water mixture Cavitation (bubbles) within the water bath Figure 4. 4: Actual image of the Ultrasonic bath (brand name: Elma) taken during laboratory session Experimental procedure: ? Clamp the process beaker so that the entire solid-liquid mixture in the beaker is subjected to the ultrasonic waves generated in the water bath. ? In order to test the effect of power, set the power knob at 100% and again at 50% while all other criteria are kept constant. . e. the frequency: 35 kHz, temperature:40oC, and another run with the frequency:130 kHz and temperature: 40oC. ? For each set of conditions, run the experiment twice. That is, for 15 minutes with samples extracted via a pipette every 3 minutes. And for 30 minutes with samples extracted every 6 minutes. ? In order to test the effect of frequency, set the frequency at 35 kHz and then again at 130 kHz while all other criteria were kept constant. i. e. the power: 100%, temperature: 40 oC , and another run with the power: 50% and temperature: 40oC 17 | P a g e H83 CEL Chemical Engineering Laboratory

Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge ? For each set of conditions, the experiment is run twice. That is, for 15 minutes with samples extracted via a pipette every 3 minutes. And for 30 minutes with samples extracted every 6 minutes. ? For the entire experiment using ultrasonic water bath, a total of 42 samples were collected. Experimental difficulties: Temperature control was the most difficult aspect of the experiment. It was very difficult to maintain the temperature at 40oC as the process proceeded. Thus the thermometer was constantly used to measure the temperature.

In order to overcome this problem, whenever the temperature exceeded 40oC the temperature was set to about 35 oC until the temperature returned to 40oC. After this, the temperature was set again at 40oC 3. Non conventional SLE extraction using Ultrasonic Horn An ultrasonic horn is a device commonly used for elevate the oscillation displacement amplitude provided by an ultrasonic transducer . The ultrasonic horn used in the laboratory is a conventional horn which is used in small scale process laboratory investigations. They are able to produce very high ultrasonic amplitudes and power densities within a liquid. Sonomechanics 2011) For the ultrasonic horn, the criterion that was being varied is the amplitude (20 % and 40 %) Figure 4. 5: Schematic of an ultrasound horn (Mason and Povey 1998; Ozcan 2006) Figure 4. 6: Schematic representation of the ultrasonic probe experimental setup (Sivakumar et al. 2006 18 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Horn tip Process beaker Ice bath Figure 4. 7: Actual image of ultrasonic probe experiment taken during laboratory session Experimental procedure: ? ? ? ? ?

Place the process beaker containing the solid-liquid mixture in a larger beaker containing ice cubes which act as a temperature controlling jacket. Immerse the horn probe into the process beaker so that 3/4th of the tip is covered with the process fluid. Set the ultrasonic amplitude to 20 % and then again at 40 % For each set of conditions, run the experiment twice. That is, for 15 minutes with samples extracted via a pipette every 3 minutes. And for 30 minutes with samples extracted every 6 minutes. For the entire experiment a total of 24 samples were taken. Experimental difficulties:

Due to the size of the ultrasonic probe, there was very littl e space to insert the pipette through the paraffin paper in order to extract the process fluid. To overcome this problem, a second hole was made in the paraffin sheet in order to take samples at the required time intervals. Also, the ice in the temperature controlling jacket kept melting; therefore it needed to be replaced. 4. UV/VIS spectrometer for sample analysis The following apparatus (brand-name: Lambda 35 UV/VIS spectrometer) is used to measure the absorbance of bio-active constituents in the samples taken during each experiment.

Experimental procedure: ? ? Pour distilled water into once square section-cell. Wipe any excess moisture on the sides of the cell and place the cell in the section reserved for the reference cell. Pour the relevant sample into another cell. Wipe thoroughly and place in the sample section. 19 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge ? ? ? Close the cover, and run the program. The anal ysis ends in approximately 30 s. The next sample for the relevant batch of samples must then be placed.

Once a certain batch of samples has been completely analyzed, the software generates a graph of absorbance against the wavelength. Figure 4. 8: (From L to R) UV/VIS spectrometer used in laboratory, view of sample and reference slots in the device. Experimental difficulties: Due to unavailability of the UV/VIS spectrometer, most samples could not be analyzed within 24 hours of collection. Some samples had to be shelved for 2 -3 days. Thus this could have affected the reliability and accuracy of the absorbance results obtained. 20 | P a g e H83 CEL Chemical Engineering Laboratory

Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge 5. 0 Results 5. 1 Results for conventional SLE – using hot plate magnetic stirrer Investigating effect on absorbance due to change in magnetic stirrer speed at a constant concentration Table 5. 1. 1: Absorbance values for SLE using hot plate stirrer at 6 rpm Speed/ rpm Concentration (g/100 ml) SLE- 15 Time/ min 15 Wavelength/ nm 255. 70 6 0. 5 SLE- 9 Time/ min 30 Wavelength/ nm 255. 70 RESULTS Absorbance (A) 1. 4327 2. 1304 2. 1777 2. 3210 2. 3598 2. 3698 Time/ min 0 3 6 9 12 15 Time/ min 0 6 12 18 24 30 Absorbance (A) 1. 937 2. 1606 2. 2826 2. 5821 2. 4215 2. 8726 Effect of time on absorbance at constant stirrer speedof 6 rpm 3. 5 Absorbance (A) 3 2. 5 2 1. 5 1 0. 5 0 0 5 10 15 20 25 30 35 Time (min) Graph 5. 1. 1: Effect of time on absorbance at constant stirrer speed of 6 rpm 21 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Table 5. 1. 2: Absorbance values for SLE using hot plate stirrer at 12 rpm Speed/ rpm Concentration (g/100 ml) SLE- 16 Time/ min Wavelength/ nm 12 0. 5 SLE- 10 Time/ min 30 Wavelength/ nm 255. 70 15 255. 70 RESULTS

Absorbance (A) 1. 5003 2. 2676 2. 3960 2. 4120 2. 4602 2. 4520 Time/ min 0 3 6 9 12 15 Time/ min 0 6 12 18 24 30 Absorbance (A) 2. 4932 2. 6412 2. 7119 2. 6861 2. 6122 2. 6320 Effect of time on absorbance at constant stirrer speed of 12 rpm 3 Absorbance (A) 2. 5 Speed 12 rpm; time=30 min 2 Speed 12 rpm; time 15 min 1. 5 1 Poly. (Speed 12 rpm; time=30 min) 0. 5 0 0 5 10 15 20 25 30 35 Poly. (Speed 12 rpm; time 15 min) Time (min) Graph 5. 1. 2: Effect of time on absorbance at constant stirrer speed of 12 rpm 22 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum

Author: W. M. Dinithi Ishara Wijetunge Effect of stirrer speed on absorbance at a constant time of 15 min 3 Absorbance (A) 2. 5 2 1. 5 Speed 6 rpm; time=15 min 1 Speed 12 rpm; time=15 min 0. 5 0 0 5 10 15 20 Time (min) Graph 5. 1. 3: Effect of change in stirrer speed on absorbance at constant time of 15 min Effect of stirrer speed on absorbance at a constant time of 30 min 3. 5 Absorbance (A) 3 2. 5 Speed 6rpm; time=30 min 2 Speed 12 rpm; time=30 min 1. 5 1 0. 5 0 0 5 10 15 20 25 30 35 Time (min) Graph 5. 1. 4: Effect of change in stirrer speed on absorbance at constant time of 30 min 3 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Comparison of all parameters 3. 5 Absorbance (A) 3 2. 5 Speed 6 rpm; time=15 min 2 Speed 12 rpm; time=15 min 1. 5 Speed 6rpm; time=30 min 1 Speed 12 rpm; time=30 min 0. 5 0 0 5 10 15 20 Time (min) 25 30 35 Graph 5. 1. 5: Effect of change in speed and change in time on the absorbance 5. 2 Results for non-conventional SLE – using an ultrasonic bath Investigating effect on absorbance due to change in power at constant frequency, temperature and concentration

Table 5. 2. 1: Absorbance values for SLE using ultrasonic bath at 50% power and 35 kHz frequency Frequency/ kHz 35 Power (%) 50 Temperature/oC Concentration (g/100 ml) SLE- 2 Time/min Wavelength/nm Time/min 0 3 6 9 12 15 40 0. 5 SLE- 3 15 255. 70 Absorbance (A) 2. 1186 2. 1793 2. 2416 2. 2314 2. 3601 2. 5028 Time/min Wavelength/nm RESULTS Time/min 0 6 12 18 24 30 30 255. 70 Absorbance (A) 1. 9262 2. 2507 2. 2846 2. 2727 2. 3452 2. 4385 24 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Table 5. 2. : Absorbance values for SLE using ultrasonic bath at 100% power and 35 kHz frequency Frequency/ kHz 35 Temperature/oC 40 Power (%) 100 Concentration (g/100 ml) 0. 5 SLE- 6 SLE- 1 Time/min 15 Time/min 30 Wavelength/nm 255. 70 Wavelength/nm 255. 70 RESULTS Absorbance (A) Time/min Time/min Absorbance (A) 1. 8367 0 1. 8491 0 3 2. 5515 6 2. 503 6 2. 5417 12 2. 8915 9 2. 4872 18 2. 8140 12 2. 5318 24 3. 1896 15 2. 7079 30 3. 7026 Effect of Change in Power on absorbance with time 4 3. 5 Absorbance (A) 3 Power 50%; time=15 min 2. 5 Power 50%; time=30 min 2 Power 100%; time=15 min 1. 5 1 Power 100%; time=30 min 0. 0 0 5 10 15 20 25 30 35 Time(min) Graph 5. 2. 1: Effect of change in power (and constant frequency of 35 kHz) 25 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Table 5. 2. 3: Absorbance values for SLE using ultrasonic bath at 50% power and 130 kHz frequency Frequency/ kHz 130 Power (%) 50 SLE- 7 Time/min 15 Wavelength/nm 257. 72 Absorbance (A) 1. 5308 1. 7888 2. 7274 1. 9867 2. 0723 2. 1998 Time/min 0 3 6 9 12 15 Temperature/oC Concentration (g/100 ml) SLE- 8 Time/min Wavelength/nm RESULTS Time/min 0 6 12 18 24 30 40 0. 30 255. 70 Absorbance (A) 1. 766 1. 8107 2. 0492 2. 0533 2. 176 2. 2159 Table 5. 2. 4: Absorbance values for SLE using ultrasonic bath at 100% power and 130 kHz frequency Frequency/ kHz 130 Power (%) 100 SLE- 4 Time/min Wavelength/nm Time/min 0 3 6 9 12 15 Temperature/oC Concentration (g/100 ml) SLE- 5 15 Time/min 255. 02 Wavelength/nm RESULTS Absorbance (A) Time/min Absorbance 1. 7263 0 1. 8567 1. 921 6 1. 903 2. 0450 12 1. 9788 2. 0523 18 2. 0763 2. 1089 24 2. 1562 2. 2202 30 2. 2983 40 0. 5 30 255. 70 (A) 26 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum

Author: W. M. Dinithi Ishara Wijetunge Effect of Change in Power on absorbance with time (Frequency 130kHz) 3 Absorbance (A) 2. 5 Power 50%; time=15 min 2 Power 50%; time=30 min 1. 5 1 Power 100%; time=15 min 0. 5 Power 100%; time=30 min 0 0 5 10 15 20 25 30 35 Time(min) Graph 5. 2. 2: Effect of change in power with time (and constant Frequency 130 kHz) Investigating change in absorbance due to change in frequency at constant power, temperature and concentration Table 5. 2. 5: Absorbance values for SLE using ultrasonic bath at 35 kHz frequency and 50% power Frequency/ kHz 35 Power (%) 50 40 Temperature/oC

Concentration (g/100 ml) SLE- 2 0. 5 SLE- 3 Time/min 15 Time/min 30 Wavelength/nm 255. 70 Wavelength/nm 255. 70 Time/min RESULTS Absorbance Time/min (A) Absorbance (A) 0 2. 1186 0 1. 9262 3 2. 1793 6 2. 2507 6 2. 2416 12 2. 2846 9 2. 2314 18 2. 2727 12 2. 3601 24 2. 3452 15 2. 5028 30 2. 4385 27 | P a g e H83 CEL Chemical Engineering Laboratory Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Table 5. 2. 6: Absorbance values for SLE using ultrasonic bath at 130 kHz frequency and 50% power Frequency/ kHz 130 Power (%) 50 40 Temperature/oC Concentration (g/100 ml) SLE- 7 0. SLE- 8 Time/min 15 Time/min 30 Wavelength/nm 257. 72 Wavelength/nm 255. 70 RESULTS Time/min Absorbance (A) Time/min 0 1. 5308 0 1. 766 3 1. 7888 6 1. 8107 6 2. 7274 12 2. 0492 9 1. 9867 18 2. 0533 12 2. 0723 24 2. 176 15 2. 1998 30 2. 2159 Absorbance (A) Effect of Change in Frequency with time (Power 50%) 3 Absorbance (A) 2. 5 Frequency 35 kHz; time=30 min 2 Frequency 35 kHz; time=15 min 1. 5 Frequency 130 kHz; time=15 min 1 Frequency 130 kHz; time=30 min 0. 5 0 0 5 10 15 20 25 30 35 Time (min) Graph 5. 2. 3: Effect of change in frequency at 50% power 28 | P a g e H83 CEL Chemical Engineering Laboratory

Solid Liquid Extraction of Ganoderma Lucidum Author: W. M. Dinithi Ishara Wijetunge Table 5. 2. 7: Absorbance values for SLE using ultrasonic bath at 35 kHz frequency and 100% power Frequency/ kHz 35 Temperature/oC 40 Power (%) 100 Concentration (g/100 ml) 0. 5 Time/min 15 Time/min 30 Wavelength/nm 255. 70 Wavelength/nm 255. 70 SLE- 6 SLE- 1 RESULTS Absorbance (A) Time/min Time/min Absorbance (A) 0 1. 8491 0 1. 8367 3 2. 5515 6 2. 503 6 2. 5417 12 2. 8915 9 2. 4872 18 2. 8140 12 2. 5318 24 3. 1896 15 2. 7079 30 3. 7026 Table 5. 2. 8: Absorbance values for SLE using ultrasonic bath at 130 kHz

x

Hi!
I'm Ruth!

Would you like to get a custom essay? How about receiving a customized one?

Check it out