Biochem Lab - Common Lab Apparatuses PDF

BIOCHEM CHAPTER 1-5 EFFICIENCY Laboratory - Prelim When you know how to use laboratory apparatuses efficiently, you can work more pro...

BIOCHEM CHAPTER 1-5 EFFICIENCY Laboratory - Prelim When you know how to use laboratory apparatuses efficiently, you can work more productively. Time and COMMON LABORATORY APPARATUSES resources are better utilized when you are skilled in Laboratories are essential hubs of scientific setting up and operating equipment. discovery and innovation. Scientists and researchers rely on these facilities to conduct experiments, analyze COST-EFFECTIVENESS data, and make groundbreaking discoveries that Mishandling laboratory equipment can lead to advance our understanding of the world. damage or breakage, resulting in the need for repairs One key aspect that contributes to the success and or replacements. Being familiar with the apparatuses efficiency of laboratory work is the knowledge and reduces the likelihood of accidents, saving on familiarity with common laboratory apparatuses. maintenance costs. The importance of understanding laboratory apparatuses extends beyond individual experiments. EXPERIMENTAL DESIGN Researchers who are well informed about the Understanding the capabilities and limitations of capabilities and limitations of different equipment can different laboratory apparatuses allows you to design design experiments more effectively. They can select experiments more effectively. You can select the most the most appropriate apparatus for specific tasks, appropriate equipment for specific tasks, ensuring the ensuring the success of their research objectives. success of your research. Proficiency in using laboratory apparatuses is also crucial for compliance with protocols and standard TROUBLESHOOTING operating procedures (SOPs.) Laboratories follow strict guidelines to maintain consistency and quality in In the event of an equipment malfunction or experimental procedures. Familiarity with the unexpected result, familiarity with the apparatuses equipment ensures researchers adhere to these enables you to troubleshoot and identify potential protocols, contributing to a standardized approach to issues more quickly. This aids in resolving problems scientific research. efficiently and minimizing downtime. Knowing and familiarizing yourself with laboratory apparatuses is of most importance in laboratory ADAPTABILITY setting for several reasons: Different laboratories may have variations of the same equipment or use slightly different models. Familiarity SAFETY with various types of apparatuses allows you to adapt Proper knowledge of the laboratory apparatuses and work comfortably in different laboratory settings. helps reduce the risk of accidents and injuries. When you understand how to handle and use equipment COLLABORATION correctly, you can minimize the chances of spills, When working with others in a laboratory, knowing breakages, and other hazardous incidents. the apparatuses promotes effective communication and collaboration. You can share knowledge and ACCURACY AND RELIABILITY techniques, facilitating teamwork and collective Familiarity with laboratory apparatuses ensures that problem-solving. you use them correctly, leading to more accurate and reliable experimental results. Precise measurements COMPLIANCE and proper usage of equipment contribute to the Many laboratory protocols and standard operating reproducibility experiments and the overall scientific procedures (SOPs) require adherence to specific validity of your work. equipment usage guidelines. Familiarity with the apparatuses ensures you comply with these protocols, maintaining consistency and quality in experimental procedures.  PROFESSIONAL DEVELOPMENT GRADUATED Used to measure volume very CYLINDER Demonstrating proficiency with laboratory precisely apparatuses showcases your skills and knowledge in LAB STAND Used to hold a variety of lab a scientific setting. This can boost your credibility and equipment open up opportunities for more complex research projects and collaborations. FUNNEL Used to pour liquids into containers with small openings.or to hold filter Knowing and familiarizing common laboratory paper apparatuses is vital for the success, safety, and efficiency of scientific research. It empowers BUNSEN Used to heat objects BURNER researchers to conduct experiments with confidence, ensuring accuracy and reliability in their results. ERLENMEYER A narrow-mouthed container used FLASK Whether it's safeguarding against accidents, to transport, heat or store promoting cost-effectiveness, or contributing to substances, often used when a advancements in scientific knowledge, the importance stopper is required of this knowledge cannot be overstated. WIRE GAUZE Used to suspend glassware over the Emphasizing the acquisition of such knowledge is Bunsen burner pivotal for nurturing a generation of competent scientists ready to tackle the challenges of tomorrow, GLASS ROD/ A skinny solid glass rod used in thus the need for students and anyone working in the STIRRING ROD chemistry to mix chemicals and laboratory to know and familiarize themselves with liquids common laboratory apparatuses. APPARATUS DESCRIPTION TEST TUBE Used to hold many test tubes filled RACK with chemicals TONGS Used to pick up or hold hot objects SAFETY Protects the eyes from flying objects GOGGLES TEST TUBE This is used to clean the inside of or chemical splashes BRUSH test tubes or graduated cylinders BEAKER A wide-mouthed container used to BEAKER TONG Used to transport a hot beaker transport, heat or store substances MORTAR AND This is used to grind substances to TEST TUBE A small glass container used to view PESTLE powder. chemical reactions or to heat small amounts of a substance GLOVES Important to wear in order to protect the skin from chemical substances. TRIPLE BEAM A device to measure the mass of an BALANCE object or substance THERMOMETER A device used for measuring the temperature of a substance. LAB COAT Protects the scientist and the scientist's clothes from hazardous or CENTRIFUGE Used to separate solids out of hot chemicals liquids that are held in test tubes by rapid rotation DROPPER Used to dispense a very small amount of a liquid EVAPORATING This is used to separate water and DISH solids from a solution by allowing STRIKER Used to light a Bunsen burner the water to evaporate off into the RING CLAMP Attaches to a lab stand and used to air hold a variety of lab equipment  6. Watch the solvent rise up the paper, carrying and SEPARATING PLANT PIGMENT COMPONENTS BY separating the pigments as it goes. PAPER CHROMATOGRAPHY 7. At the instant the solvent reaches to the top, remove the paper and let it dry. Chlorophyll has a tendency to conceal the presence 8. Observe the bands of pigment. Identify and label of other pigments in leaves. However, during autumn, the pigment bands on the dry strip. chlorophyll disintegrates, allowing xanthophyll, 9. Each of the pigments will have an Rf value, which carotene, and newly formed anthocyanin to reveal refers to the speed at which it moves over the paper their distinct colors. To analyze the mixture of compared with the speed of the solvent. This can be pigments in a leaf, a method called paper acquired by dividing the distance traveled by the chromatography is employed pigments by the distance traveled by the solvent. CHROMATOGRAPHY BUFFER SOLUTION PREPARATION AND PH MEASUREMENT Is a process of separating mixtures into individual components and is literally translated as "color writing." BUFFER In this technique, the components of the mixture in a A buffer, in the context of chemistry, refers to a liquid medium are separated based on absorption and solution that resists changes in its pH level when an capillarity. acidic or basic substance is added to it. The paper serves as a medium for absorption, and Buffers play a crucial role in maintaining the stability capillarity draws the substances up the paper at and proper functioning of various biological, chemical, different rates. and industrial processes. As a result, the pigments get separated on the When an acidic substance is introduced into a buffer paper and are visible as colored streaks. The resulting solution, the weak base component of the buffer pattern of separated components on the paper is reacts with the additional H+ ions (hydrogen ions) to referred to as a chromatogram. neutralize them, preventing a significant change in pH. Conversely, if a basic substance is added to the PROCEDURE buffer, the weak acid component reacts with the OH- 1. Cut a strip of filter paper or chromatography paper ions (hydroxide ions) to maintain the pH within a so that it just fits inside a 15 cm test tube. Cut a point relatively narrow range. at one end, then draw a pencil line just near the Buffers are essential in many biological systems, pointed end of the paper. Prepare 3 setups of this. such as in blood, where they help maintain a constant 2. Tear 3 different types of plant leaves into smaller pH level necessary for the proper functioning of pieces. Put each type of leaves into separate mortars. enzymes and other biomolecules. In our blood, Add about 5 mL ethyl alcohol to the leaf pieces. carbonic acid is the most important buffer. This 3. Crush leaves with the pestle, using a circular solution maintains our blood pH (7.35- 7.45) to motion, until the mixture is finely ground. The liquid facilitate transport of oxygen from the lungs to the formed is now called the pigment extract. cells. 4. Use a glass rod to touch a drop of the pigment They are also utilized in chemical laboratories to extract to the center of the pencil line on the paper stabilize reactions by maintaining a constant pH strip. Repeat as many as 20 times, to the pigment environment. In industrial processes, buffers are often spot. Remember to let the dot dry after each drop is used to control pH levels and ensure the efficiency of added. The drying keeps the pigment dot from chemical reactions. spreading out and builds up too much. 5. Pour 5 mL chromatography solvent into the test tube. Fit the paper inside, holding it steadily so that it does not touch the sides of the tube. Adjust the paper so that the point just touches the solvent. The pigment dot must be above the level of the solvent.  PROCEDURE MONOSACCHARIDES These are the simplest form of carbohydrates and MEASUREMENT OF PH cannot be further broken down into smaller sugars. 1. Get 7 test tubes. They consist of a single sugar unit. 2. Put 1 mL of 0.1M HCl in the first test tube and dip a Common monosaccharides include glucose (also pH paper. known as dextrose), fructose, and galactose. 3. Repeat the procedure with 0.1M acetic acid, 0.1M sodium acetate, 0.1M carbonic acid, 0.1M sodium DISACCHARIDES bicarbonate, 0.1M ammonia, and 0.1M sodium Disaccharides are composed of two monosaccharide hydroxide. units linked together by a glycosidic bond. They are 4. Record your results in your notebook. formed when two monosaccharides undergo a 5. If a pH meter is available, determine the pH of the condensation reaction, releasing a water molecule. solutions listed above. Record your results and Examples of disaccharides are sucrose (glucose compare them with the result you obtained in using +fructose), lactose (glucose +galactose), and maltose the pH paper. (glucose +glucose). BUFFER SYSTEM POLYSACCHARIDES 1. Get four 50 mL beakers. Label them as A, B, C and Polysaccharides are complex carbohydrates made D. Prepare buffer systems in four beakers as follows: up of long chains of monosaccharide units linked a. 10 mL 0.1M CH3COOH (Acetic Acid) + 10mL together. These chains can be branched or M 0.1NaC2H3O2 (Sodium Acetate) unbranched. b. 5 mL 0.1M CH3COOH (Acetic Acid) + 15 mL Polysaccharides serve as a storage form of energy in M 0.1 NaC2H3O2 (Sodium Acetate) plants and animals and also play structural roles in c. 10 mL 0.1 M H2CO3 (Ammonium Chloride) + cells and tissues. 10 mL 0.1 NaHCO3 (Ammonia) Some common polysaccharides include starch d. 5 mL 0.1 M H2CO3 (Ammonium Chloride) + (found in plants), glycogen (stored in animals, 15 mL 0.1 NaHCO3 (Ammonia) including humans), and cellulose (a major component 2. Measure the pH of each buffer system using either of plant cell walls). pH paper or pH meter. Record your results. 3. Divide each of the buffers (A-D) into halves (10mL CARBOHYDRATES each) and place half of each in a clean, dry 50 mL beaker. Carbohydrates are an important source of energy 4. To the first half, add 1mL of 0.1 M HCL. Mix and because they can be easily broken down during determine the pH. To the second half add 1 mL of 0.1 digestion into glucose, which is used by cells to M NaOH. Mix and determine the pH. produce adenosine triphosphate (ATP), the primary 5. Record your answers. energy currency of the body. However, not all carbohydrates are equal in terms of TEST FOR CARBOHYDRATES: nutritional value. Simple carbohydrates, such as IODINE AND SELIWANOFF'S TEST those found in sugary snacks and processed foods, can lead to rapid spikes in blood sugar levels and are generally considered less healthy. CARBOHYDRATES On the other hand, complex carbohydrates found in Carbohydrates are a class of organic compounds whole grains, vegetables, and legumes provide a composed of carbon, hydrogen, and oxygen atoms. more sustained release of energy and are often a They are one of the essential macronutrients found in better choice for maintaining a balanced diet. various foods, providing a major source of energy for living organisms, including humans. Carbohydrates can be divided into three main categories based on their chemical structure:  TESTS FOR CARBOHYDRATES DETERMINING PURITY IN Tests for carbohydrates serve several purposes, BIOCHEMICAL RESEARCH mainly in the fields of biology, biochemistry, and food In biochemistry, it is essential to ensure the purity of science. carbohydrate samples used in experiments. Various These tests are designed to detect the presence of tests help researchers assess the quality and quantity carbohydrates in various substances and to of carbohydrates isolated or synthesized for research characterize their properties. Some common reasons purposes. for conducting carbohydrate tests include: ENVIRONMENTAL AND IDENTIFYING THE PRESENCE OF CARBOHYDRATES INDUSTRIAL APPLICATIONS Carbohydrate tests are used to determine whether Carbohydrate tests find applications in carbohydrates are present in a given sample or environmental monitoring, such as assessing the biological material. This information can be crucial for presence of organic matter in water samples. They are understanding the composition of food products, also used in industrial processes, like determining the biological fluids, or tissues. sugar content during fermentation in the production of alcoholic beverages. NUTRITIONAL ANALYSIS Carbohydrates are a primary source of energy in the Overall, carbohydrate tests play a critical role in human diet. Food scientists and nutritionists use understanding the role of carbohydrates in biological carbohydrate tests to quantify the amount of systems, evaluating their nutritional significance, and carbohydrates in foods, which helps in evaluating their ensuring the quality and safety of various products in nutritional content and caloric value. multiple fields of science and industry. QUALITY CONTROL IN THE FOOD INDUSTRY Below are common tests/ experiments conducted to determine the presence/ absence and types of Food manufacturers use carbohydrate tests to carbohydrates present in a sample. For each of the ensure that their products meet certain standards and tests, determine the purpose of the test and the specifications. For example, checking sugar levels in principle behind the test, then perform the steps that fruit juices or determining starch content in processed follow. foods. PROCEDURE MEDICAL DIAGNOSIS Carbohydrate tests are utilized in clinical settings to A. IODINE TEST FOR POLYSACCHARIDES diagnose and monitor certain medical conditions. For 1. Take 2mL of each of the samples in separate test instance, blood glucose tests are commonly used to tubes and take 2mL of distilled water in another tube diagnose diabetes and monitor blood sugar levels ni as control. diabetic patients. 2. Add 2-3 drops of iodine solution to all the tubes 3. Observe the appearance of color in the test tubes, CARBOHYDRATE METABOLISM STUDIES and record them on your notes. Carbohydrate tests are valuable in research related 4. Heat the test tubes in the water bath until the color to carbohydrate metabolism. Scientists use these tests disappears. to investigate how the body processes carbohydrates 5. Take the test tubes out for cooling. Observe what and to study various metabolic disorders. happens after the test tube has cooled down.  B. SELIWANOFF'S TEST BARFOEDS’S TEST 1. Take clean, dry test tubes and add 1mL of each of The Barfoed's test is a chemical test used to the test samples and 1mL of distilled water in another distinguish between monosaccharides and certain as blank. disaccharides. 2. Add 2mL of Seliwanoffs' reagent to all the test Specifically, it helps to differentiate between tubes. reducing monosaccharides (those that have a free 3. Heat the solution in a boiling water bath for two aldehyde or ketone group) and non-reducing minutes. disaccharides (those that lack a free aldehyde or 4. Observe the formation of color and note it down ketone group) like sucrose. During the Barfoed's test, the sample containing the TEST FOR CARBOHYDRATES: sugar is mixed with Barfoed's reagent, which is a BENEDICT AND BARFOED'S TEST solution of copper acetate in acetic acid. The mixture is then heated in a boiling water bath for a specified period, usually 2-3 minutes. BENEDICT’S TEST If the sugar is a reducing monosaccharide, such as Benedict's test is a chemical test used to detect the glucose or fructose, it will react with copper ions in presence of reducing sugars in a solution. Barfoed's reagent. Reducing sugars are a class of sugars that have a This reaction leads to the reduction of copper ions to free aldehyde or ketone functional group and can form a brick-red precipitate of copper (I) and oxide donate electrons to other compounds, reducing them (Cu2O).The intensity of the color will vary depending in the process. on the amount of reducing sugar present. Examples of reducing sugars include glucose, On the other hand, non-reducing disaccharides like fructose, maltose, and lactose. sucrose will not react with Barfoed's reagent and, In the Benedict's test, the reducing sugar is mixed therefore, will not produce a brick-red precipitate. with Benedict's reagent, which is a solution of copper sulfate (CuSO4) and sodium citrate in a strong PROCEDURE alkaline medium (usually sodium carbonate or sodium hydroxide). The mixture is then heated in a boiling water bath for a few minutes. Benedict's Test If the solution contains reducing sugars, they will 1. Get the 6 different sugars prepared by your lab react with the copper ions in the Benedict's reagent. instructor. This reaction results in the reduction of the copper 2. Take 5 mL of Benedict's qualitative reagent in 6 ions to form a red or reddish-brown precipitate of separate test tubes. copper oxide (Cu2O). The intensity of the color can 3. In each test tube with the Benedict's reagent, add 8 vary depending on the amount of reducing sugar drops of the prepared sugars. present in the solution. 4. Mix the solutions well. Benedict's test is commonly used in biochemical and 5. Hold the test tube on flame and boil for 2 minutes. food analysis to determine the presence and 6. Allow the solution to cool. approximate concentration of reducing sugars in 7. Look for the precipitates. various samples, such as urine, blood, and certain food products. Barfoed's Test It is a simple and widely used test for the qualitative 1. Get the 6 different sugars prepared by your lab detection of reducing sugars. instructor. 2. In separate test tubes, put 1 mL each of the sugars. 3. Add 2 mL Barfoed's solution. 4. Boil the tubes in a boiling water bath for 2 minutes. 5. Observe the presence of precipitates in the test tube. 6. Record the color of the precipitates formed