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VIETNAM NATIONAL UNIVERSITY – HO CHI MINH CITY INTERNATIONAL UNIVERSITY SCHOOL OF BIOMEDICAL ENGINEERING Chemistry for BME Laboratory BM098IU REPORT LAB 4: SEPERATION METHOD Submitted by Hồ Anh Khoa – BEBEIU20168 Date Submitted: Date Performed: Lab Section: Course Instructor: TAs: 09/12/2022 02/12/2022 Friday Morning Truong Phuoc Long, PhD Tran Duong An Hoa Dao Ngoc Yen Khoa Nguyen Manh Khang GRADING GUIDELINE FOR LAB REPORT Number Content Score Comment Format (max 10%) - Font type Yes No - Font size Yes No - Lab title Yes No - Page number Yes No - Table of contents Yes No - Header/Footer Yes No - List of figures (if exists) Yes No - List of tables (if exists) Yes No - Lab report structure Yes No - References Yes No English Grammar and Spelling (max 5%) (Max 0% if references = 0%) - Grammar Yes No - Spelling Yes No Introduction: Data and Result Analysis (max 85%) 05 parts, 17% for each Experimental procedure: Results & Discussion: Assignment: Total Score Conclusion: Overall Signature _ International University i School of Biomedical Engineering Date: …/…/2022 BM098IU Table of Contents I Introduction 1 What is Separation? a) Separation by distillation b) Separation by extraction 2 Essential oil of Pomelo peel Tannin II Experimental procedures Materials and equipment a Materials b Equipment Experimental procedures a Experiment 1: Essential oil extraction from pomelo peel by hydro-distillation method b Experiment 2: Tannins extraction of tea leaves III Results and discussion Results a Essential oil extraction from pomelo peel by hydro-distillation method b Tannins extraction from tea leaves Discussion a Essential oil extraction from pomelo peel by hydro-distillation method b Tannins extraction from tea leaves IV Assignment 10 V Conclusion… 12 VI References 12 International University ii School of Biomedical Engineering BM098IU List of Figures Figure Distillation method Figure Pomelo peel Figure Epigallocatechin gallate Figure Ultrasonic P Elma Figure Water bath Figure Set up for experiment Figure Tannins after the filtration contained in the beaker Figure Filter the mixture Figure Filter paper folding technique Figure 10 Extracting essential oil from pomelo peel is contained in amber glass bottles Figure 11 Filtered tannins contained in test tubes Figure 12 Tannins extraction from tea leaves International University iii School of Biomedical Engineering BM098IU International University School of Biomedical Engineering BM098IU I Introduction In this lab we learn about three separation methods based on boiling point, density and solubility as well as the equipment used in separation After that, I was also guided in practical experiments related to each process: Using the hydro-distillation method to extract essential oil from pomelo peel, tannins extraction of tea leaves Background information: What is Separation? A separation is a technique that divides a chemical combination or solution into two or more different product mixes At least one of the separation products is a supplement in one or more of the elements of the source mixture A separation technique may separate a mixture into its pure ingredients in some situations Separations take use of variations in chemical or physical characteristics (Separation: Definition, methods, complete and incomplete TOPPR-guides) a) Separation by distillation Distillation is a separation process used to separate a solid from a liquid mixture It is essentially the process of heating a liquid to produce vapors and then condensing the vapors to recover the liquid The liquid obtained by undergoing vapor condensation is known as distillate Miscible liquids combine to produce a solution (Separation by distillation 2021) Figure Distillation method International University School of Biomedical Engineering BM098IU b) Separation by extraction The transferring of compound(s) from a solid or liquid into a different solvent or phase is referred to as extraction The most typical method in the chemical lab is liquid-liquid extraction, which takes place in a separatory funnel In the funnel, a dissolved component solution is put, and an immiscible solvent is added, resulting in two layers that are shaken together It is usual for one layer to be aqueous and the other to be organic Components are "extracted" when they are moved from one layer to the next Extraction is used in the chemical lab for a variety of reasons It is the most often used method for separating chemicals from plant sources Extraction is the process of transferring chemicals from one liquid to another so that they can be more readily controlled or concentrated It also allows for the selective elimination of mixed components (Libretexts, 4: Extraction 2020) Essential oil of Pomelo peel Essential oils derived from pomelo peels are widely used in a variety of applications and have been identified as antibacterial and antioxidant alternatives It has been demonstrated that the bioactivity of essential oils is determined by their chemical makeup Pomelo peel contains a significant quantity of essential oil, around 0.4-1% of the peel's weight; this essential oil comprises a variety of volatile aromatics Pomelo essential oil is often composed of limonene (80-88%), -Pinene (0.8-1.2%), linalool (1.1.-0.7%), and terpinene (0.7-1.0%) Pomelo essential oil exhibits antibacterial properties against Staphylococcus aureus, Enterococcus faecalis, Staphylococcus epidermidis, Escherichia coli, Salmonella typhimurium, and Proteus vulgaris, according to research Essential oils are popular due to their health, beauty, and relaxation advantages They not only deliver a nice perfume but also assist decrease tension exceptionally efficiently by dispersing the aroma of essential oils to the outside, producing a fresh, clean area free of bacteria and germs Furthermore, the use of essential oils in aromatherapy reduces the usage of chemicals that can harm human health (Ngan et al., 2021) International University School of Biomedical Engineering BM098IU Figure Pomelo peel Tannin Tannins are a type of chemical compound that is part of a wider group of chemicals known as polyphenols Tannin particles are usually bigger than other groups of polyphenol chemicals Tannins may readily interact with other molecules such as proteins and minerals Tannins are naturally present in a variety of food and non-edible plants, including bark, leaves, spices, nuts, fruits, and legumes Tannins are chemicals found in plants that aid in insect control Tannins also add color and taste to a variety of plant meals Tea, coffee, wine, and chocolate are some of the richest and most prevalent sources of tannins The bitter flavor of these meals and beverages is generally attributable to their high tannin content Although tea is commonly thought to be a high-tannin source, the amount of tannins in tea varies depending on a variety of circumstances The four basic varieties of tea are white, black, green, and oolong, and they are all brewed from the leaves of the Camellia sinensis plant Tannins are present in all teas, although the concentration is heavily controlled by how and for how long the tea leaves are processed According to some sources, black tea contains the greatest concentration of tannins of any tea variety, whereas green tea has the lowest Tannin levels in white and oolong teas are typically International University School of Biomedical Engineering BM098IU medium, although the quantity in each can vary significantly depending on how the tea is manufactured Epigallocatechin gallate is one of the primary tannins present in green tea (EGCG) EGCG belongs to a class of chemicals known as catechins, which are thought to be one of the causes behind green tea's numerous health advantages (What are the tannins in tea and what are their benefits?) Figure Epigallocatechin gallate II Experimental procedures Materials and equipment a Materials • Pomelo peels • Tea leaves • Distilled water • Na2SO4 • 70 Ethanol • FeCl3 b Equipment • Test tube International University School of Biomedical Engineering BM098IU • Round-bottom flask • Beaker • Micropipette • Mortar and pestle • Heating mantle • Analytical balance • Ultrasonic P Elma • Water bath • Filter paper and funnel • Volumetric pipette • Cylinder Figure Water bath Figure Ultrasonic P Elma Experimental procedures a Experiment 1: Essential oil extraction from pomelo peel by hydrodistillation method International University School of Biomedical Engineering BM098IU Step 1: Weigh and cut 250 g of pomelo peel, then transfer to a L round-bottom flask with 500 mL of distilled water Step 2: Place the flask on a heating mantle then connect the still column and conde nser to the round bottom flask Step 3: Turn on the heating mantle and set the appropriate boiling point Once it starts to boil, turn the heat down Step 4: After completing the process of water-bath distillation, the essential oil is obtained, separated and anhydrous with Na2SO4 Step 5: Calculate the Yield of pomelo peels oil (Y) using the formula below: Y (%) = Volume of essential oil (mL) Amount of raw material (g) × 100 Step 6: Store essential oils in amber glass bottles and store at 4°C for further experiments Figure Set up for experiment b Experiment 2: Tannins extraction of tea leaves Step 1: Preparation of tea leaves Cut the tea leaves into small pieces Then put the chopped tea leaves on the mortar and pound with a pestle The experimenter must prepare 5g of small International University School of Biomedical Engineering BM098IU tea leaves divided equally into beakers by analytical balance and label beakers from to Step 2: For the first beakers, add 100 mL of distilled water For the last beaker, add 100mL of 70 ethanol Step 3: Divided into cases: Place the first beaker in water bath at 100°C, the second and fourth at room temperature and the third beaker in ultrasonic cleaner for 40 minutes Figure Tannins after the filtration contained in the beaker Step 4: After 40 minutes, use a funnel and filter paper to filter the mixture in each beaker Figure Filter the mixture International University School of Biomedical Engineering BM098IU Figure Filter paper folding technique Note: • Make sure you use the right size filter paper • The size of the paper should be less than that of the bottom of the funnel (It must cover all the holes) • Filter paper must have no creases or folds Step 5: Use a volumetric pipette to draw mL of the tannin solution and add to test tubes Then add mL FeCl3 to each test tube Observe the result in each test tube The color turns blackish or bluish, it indicates the presence of tannin III Results and discussion Results a Essential oil extraction from pomelo peel by hydro-distillation method Figure 10 Extracting essential oil from pomelo peel is contained in amber glass bottles International University School of Biomedical Engineering BM098IU b Tannins extraction from tea leaves Figure 11 Filtered tannins contained in test tubes Figure 12 Tannins extraction from tea leaves Discussion a Essential oil extraction from pomelo peel by hydro-distillation method In the experiment, we obtained about mL in 100g of tea leaves The Yield of pomelo peels oil: Y (%) = mL 100 g × 100 = 2% b Tannins extraction from tea leaves International University School of Biomedical Engineering BM098IU In this experiment, a distinct difference in the color of tannin extracted from tea leaves could be observed in each test tube For test tube 1, the color changed from green to light black indicating the presence of tannin but very little For the remaining three test tubes, the color turned black clearly indicating that the tannin content contained in the test tubes was very high IV Assignment What is the principle of distillation? What are the applications of distillation? • Principle of distillation: The distillation process is based on the fact that different liquids boil at different temperatures When a combination is heated, the material with the lower boiling point begins to boil first, converting into vapors that may then be collected separately • Applications of distillation: - Recovery of Solvents - Extraction of the Volatile Oils - Manufacturing of official preparations - Purification What is the difference between distillation and fractional distillation? Boiling Point Fractional Distillation: Fractional distillation is used to separate liquids with similar boiling points Simple Distillation: This method is used to separate liquids having boiling point differences of at least 50 degrees • Apparatus Fractional Distillation: A complicated apparatus with a fractionating column is used in fractional distillation 10 International University School of Biomedical Engineering BM098IU Simple Distillation: A simple apparatus is employed, consisting of a flask to retain the mixture, a condenser, and a flask to collect the purified components • Repetition Fractional Distillation: The procedure must be performed multiple times to obtain the pure components Simple distillation is a one-time procedure • Solvent separation from a Solute Fractional distillation is ineffective for separating a solvent from a solute Simple distillation is a method for separating a solvent from a solute (Pabasara, 2017) Which factors affect the rate of sedimentation of a particle during centrifugation? Please explain the reasons The centrifugal sedimentation method determines particle size by changing the speed of particles moving due to centrifugal force When a rotational force is applied to particles, bigger particles settle quicker and smaller particles settle slower - Factors affect the rate of sedimentation of a particle during centrifugation • Density: The centrifuge's spinning causes more dense particles to move to the outer edge • Temperature: The energy contained within a spinning centrifuge raises the temperature within the centrifuge • Distance of particles displacement: As the particles' distance from the center of rotation rises, so does their relative centrifugal force • Speed: As the particles in the band travel through the density medium, zones of equal size emerge as the quicker sedimenting particles move ahead of the slower ones When extracting, using dry leaves or fresh leaves can get more essential oil? Please explain the reasons 11 International University School of Biomedical Engineering BM098IU In the drug product cell is semi-permeable, so when the leaf is fresh, due to the action of the protoplast, only the solvent is permeated into the cell, causing the drug product to expand, and the solute in the cell cannot diffuse out Therefore, for fresh leaves, in the extract, one must find a way to destroy the primary material of fresh medicinal materials that need to be dipped in alcohol at high temperature or high alcohol content, the primary membrane is broken, facilitating the passage of solutes through the cell membrane How can different gases be separated from the atmosphere? Fractional distillation: Fractional distillation is a kind of distillation that involves the separation of liquid phases This procedure requires numerous distillations and condensations, and the mixture is typically separated into its constituent elements The separation occurs when the mixture is heated to a temperature where portions of the mixture begin to evaporate (How can we obtain different gases from air? - byju's) V Conclusion In this lab, I got a clear and detailed understanding of three different separation methods based on boiling point, density, and solubility as well as the equipment used in separation I then ran them through real-world experiments involving each process: A hydro-distillation process is used to extract essential oils from pomelo peel and tannins from tea leaves VI References Ngan, T T K., Tran, T H., Minh, L T N., Long, H B., & Le, X T (2021) Application of pomelo essential oil ( Citrus Grandis L ) in effective scenting of diffused products E3S Web of Conferences, 306, 04020 https://doi.org/10.1051/e3sconf/202130604020 Separation: Definition, methods, complete and incomplete - TOPPR-guides (n.d.) Retrieved December 15, 2022, from https://www.toppr.com/guides/chemistry/is-matteraround-us-pure/separation/ 12 International University School of Biomedical Engineering BM098IU Separation by distillation GeeksforGeeks (2021, July 21) Retrieved December 15, 2022, from https://www.geeksforgeeks.org/separation-by-distillation/ Libretexts (2020, August 13) 4: Extraction Chemistry LibreTexts Retrieved December 15, 2022, from https://chem.libretexts.org/Courses/SUNY_Oneonta/Chem_221%3A_Organic_Chemistr y_I_(Bennett)/2%3ALab_Textbook_(Nichols)/04%3A_Extraction What are the tannins in tea and what are their benefits? vinmec.com (n.d.) Retrieved December 15, 2022, from https://www.vinmec.com/vi/news/health-news/nutrition/whatare-the-tannins-in-tea-and-what-are-their-benefits/ Pabasara (2017, February 27) Difference between fractional distillation and simple distillation: Process, apparatus, uses Pediaa.Com Retrieved December 15, 2022, from https://pediaa.com/difference-between-fractional-distillation-and-simple-distillation/#ef How can we obtain different gases from air? - byju's (n.d.) Retrieved December 15, 2022, from https://byjus.com/question-answer/how-can-we-obtain-different-gases-from-air/ 13 ...GRADING GUIDELINE FOR LAB REPORT Number Content Score Comment Format (max 10%) - Font type Yes No - Font size Yes No - Lab title Yes No - Page number Yes No - Table of... temperature and the third beaker in ultrasonic cleaner for 40 minutes Figure Tannins after the filtration contained in the beaker Step 4: After 40 minutes, use a funnel and filter paper to filter the... scenting of diffused products E3S Web of Conferences, 306, 040 20 https://doi.org/10.1051/e3sconf/2021306 040 20 Separation: Definition, methods, complete and incomplete - TOPPR-guides (n.d.) Retrieved