Hospital Laboratory Safety Procedures PDF

## Experiment/Activity No. 1 ### Aim To understand and apply safety measures in a hospital laboratory with biological and chemical risks associated with biological hazards, ensuring a safe working environment for laboratory personnel and patients. ### Requirements - Personal Protective Equipment (...

## Experiment/Activity No. 1 ### Aim To understand and apply safety measures in a hospital laboratory with biological and chemical risks associated with biological hazards, ensuring a safe working environment for laboratory personnel and patients. ### Requirements - Personal Protective Equipment (PPE): lab coat, gloves, safety goggles, face masks, face shields, safety spill kit, fire extinguisher, first aid kit, spill kit - Safety equipment: fume hood, sharps container, biohazard waste container - Disinfection materials: 70% ethanol or isopropyl alcohol, autoclave, disinfectant (e.g. bleach) - Other materials: material safety data sheets, labels for chemicals and biological samples, sterile containers for biological specimen handling. ### Principle Hospital laboratories deal with biological samples, chemicals and various instruments that can pose risks to health and safety. Effective safety protocols and proper techniques are essential to avoid infections, contamination and accidents. The practical focus on the principles of infection control, chemical safety, equipment handling and emergency preparedness. ### Procedure #### 1. Personal Protective Equipment (PPE) - **Lab coat:** Always wear a clean lab coat to protect your skin and clothes from exposure to chemicals, biological agents, or other hazards. - **Gloves:** Put on disposable gloves before handling any biological sample, chemical or hazardous material. Replace gloves between tasks or if damaged. - **Safety goggles:** Use safety goggles to protect your eyes from splashes of chemicals or biological fluids. - **Face masks/Shields:** Wear a mask or face shield, especially when working with infectious materials, to prevent exposure to airborne pathogens. #### 2. Infection Control Measures - **Hand hygiene:** Wash hands thoroughly with soap and water before putting on gloves, and immediately after removing them. Use hand sanitizer when soap and water are not available. - **Sterilization:** Sterilize equipment and surfaces using an autoclave, disinfectant (e.g. bleach) or 70% alcohol to kill microorganisms. - **Biohazardous Waste Disposal:** Dispose of all biohazardous materials such as used gloves, pipettes and sample containers in biohazard waste bins. Ensure the waste is properly labeled for safe disposal. - **Sharps Disposal:** Place all sharp instruments (e.g. needles, scalpels) in puncture resistant sharps containers immediately after use to avoid injury. #### 3. Chemical Safety Measures - **Labeling:** Ensure all chemicals are clearly labeled with their name, concentration, and any relevant hazard symbols. Never use unlabeled containers. - **Storage:** Store chemicals according to the hazard class (e.g. acids, bases, flammables). Ensure proper ventilation when working with volatile chemicals. - **MSDS:** Refer to the Material Safety Data Sheets (MSDS) for each chemical to understand its hazard, safe handling instructions, and emergency procedures in case of exposure or spills. #### 4. Spill Management In case of a chemical spill, isolate the area and clean it up immediately using a spill kit. Wear appropriate PPE and dispose of the waste safely. #### 4. Handling of Biological Specimens - **Sample handling:** Handle all biological samples (e.g. blood, urine, tissue) with care, assuming they are potentially infectious. - **Transport:** Place samples in leak-proof, sterile containers and tools. Use sealed containers labeled with patient information and biohazard symbols. Avoid unnecessary handling to minimize the risk of contamination. - **Waste Disposal:** Dispose of biological waste following hospital regulations. Never mix biological waste with regular trash. #### 5. Fire and Electrical Safety - **Fire Extinguisher:** Familiarize yourself with the location and type of fire extinguishers. - **In case of a fire, follow the PASS procedure:** Pull the pin, aim at the base, squeeze the handle, sweep side to side. - **Fire Safety:** Keep flammable materials away from open flames. Ensure that heat sources and other heating devices are turned off when not in use. - **Electrical Equipment:** Inspect electrical equipment for damaged cords or faulty wiring before use. Do not operate equipment with wet hands, and avoid overloading electrical outlets. #### 6. Emergency Procedures - **First Aid:** Familiarize yourself with the location of first aid kits and emergency equipment. If an accident occurs, report it immediately and follow first aid procedures. - **Post-Exposure Protocols:** If exposed to biological agents (e.g. through a needle stick or splash), wash the affected area with soap and water. Report the incident and follow hospital post-exposure protocols. - **Spill Response:** For spills involving chemicals or biological materials, contain the spill, use the appropriate spill kit, and alert your supervisor. #### 7. Equipment Safety - **Centrifuge:** Ensure centrifuge is balanced before use. Never open the lid while the rotor is moving. Regularly inspect for wear and tear. Follow hospital guidelines for autoclave operation! Ensure that items are packaged correctly and that the autoclave runs for the appropriate time at the correct temperature. - **Microscope and Diagnostic Equipment:** Clean and disinfect diagnostic equipment regularly to prevent cross-contamination. #### Observation - Check that all chemicals and biological samples are properly labeled and stored correctly. - Observe the correct handling of biological and chemical waste disposal. - Inspect electrical equipment and verify that fire safety equipment is easily accessible. #### Conclusion This practical demonstrates that strict adherence to safety measures in a hospital laboratory is essential to preventing accidents, contamination and exposure to hazardous materials. Using PPE, appropriate handling of biological materials, following infection control protocols, handling chemicals with care and being prepared for emergencies are all vital components of a safe hospital laboratory environment. ## Experiment/Activity No. 2 ### Aim To understand and practice the responsibilities of a medical laboratory technician in performing common laboratory tasks such as specimen handling, testing, and result reporting with a focus on accuracy, patient safety and quality control. ### Principle A medical lab technician plays a crucial role in the healthcare system by performing various diagnostic tests. Proper handling of specimens, adherence to standard operating procedures (SOPs), and quality control are essential to ensure accurate and reliable test results. These results guide physicians in diagnosing and managing diseases. ### Materials / Equipment 1. Urine sample collection container (for urinalysis) 2. Urine test strips (for chemical analysis) 3. Microscope (for microscopic examination) 4. Centrifuge (for urine sediment preparation) 5. Personal Protective Equipment (PPE): gloves, lab coat, face mask 6. Labeling materials (for proper sample identification) ### Procedure #### 1. Pre Analytical Responsibilities - **Patient Identification:** Confirm the patient’s identity using at least two unique identifiers (e.g. name and date of birth) to avoid sample mix up. - **Specimen Collection:** Instruct the patient on the proper method of collecting a midstream urine sample to prevent contamination. Ensure that the urine sample is collected in a clean, sterile container. - **Specimen Labeling:** Accurately label the sample container with the patient’s information (name, date and time of collection). - **Sample Handling:** Ensure that the urine sample is delivered promptly to the laboratory and stored at the appropriate temperature if not immediately processed. #### 2. Analytical Responsibilities - **Physical Examination:** Record the color and clarity of the urine sample (e.g. pale yellow, clear, or cloudy). - **Chemical Examination:** Use test strips to check for parameters such as pH, glucose, protein, ketones, nitrites and leukocyte esterase. Follow the manufacturer’s instructions and SOPs for test strip handling. Compare the test strip results with the reference chart and document findings accurately. - **Microscopic Examination (if indicated):** Centrifuge a portion of the urine sample to prepare sediment. Place a drop of the sediment on a slide and examine it under a microscope. Look for red blood cells (RBCs), white blood cells (WBCs), epithelial cells, bacteria, and crystals. Record the findings systematically, especially if abnormal components are present. - **Quality Control:** Perform routine quality control tests to ensure the reagents (e.g. test strips) and instruments (e.g. microscope, centrifuge) are functioning properly. Record quality control data, and follow corrective action if QC tests fall outside the acceptable range. #### 3. Post Analytical Responsibilities - **Result Verification:** Review the test results to ensure they are complete and accurate. Double check any abnormal findings and repeat tests if necessary. - **Documentation:** Accurately record all test results in the laboratory information system (LIS). Ensure that all patient information and test data are recorded clearly and correctly. - **Result Reporting:** Report test results to the relevant healthcare provider in a timely manner. Maintain patient confidentiality and ensure that sensitive data are handled securely. - **Sample Disposal:** Dispose of the sample and any testing materials (e.g. used test strips, slides) in accordance with biohazard waste protocols. #### Result Physical Examination: Color - Pale yellow, clarity - clear. #### Chemical Examination pH: 6.0, Protein: Negative, Glucose: Negative, Leukocyte esterase: Negative, Nitrites: Negative. #### Microscopic Examination No RBCs, no WBCs, no bacteria, and no crystals seen. #### Quality Control - Test strips were validated as functional, no major abnormalities detected. The results show a normal urinalysis with no indication of infection or other abnormalities. The technician’s responsibility in ensuring correct specimen handling, accurate testing, and reliable reporting was successfully demonstrated. #### Conclusion This practical highlights the key responsibilities of a medical laboratory technician including proper sample collection, accurate testing, and reliable reporting; adherence to laboratory safety procedures, attention to quality control, careful documentation, and ensuring that test results are accurate, timely and useful for patient care. ## Experiment/Activity No. 3 ### Aim To prepare dilute sulfuric acid (H₂SO₄) from concentrated sulfuric acid and to describe its exothermic nature. ### Requirements - Concentrated sulfuric acid (H₂SO₄) - Distilled water - Measuring cylinder - Beaker (500mL) - Glass rod - Thermometer - Dropper - Protective gear: (gloves, safety goggles, lab coat) ### Theory Sulfuric acid (H₂SO₄) is a strong, corrosive acid that is widely used in industrial processes and laboratories. It is important to handle it carefully, especially when diluting it due to its highly exothermic reaction with water. The dilution of concentrated sulfuric acid should be done by adding acid to water, not the other way around, to avoid splashing and minimize the risk of injury. ### Procedure #### 1. Preparation of Apparatus Wear safety goggles, gloves, and a lab coat before starting the experiment. Ensure that the glassware (beaker, measuring cylinder, etc.) is clean and dry. #### 2. Adding Water Measure 200mL of distilled water using the measuring cylinder and pour it into a 500mL beaker. #### 3. Dilution of Concentrated Sulfuric Acid Using a dropper, carefully add 20mL of concentrated sulfuric acid slowly into the water in the beaker. Always add acid to water, not water to acid. Stir the solution gently with a glass rod as you add the acid to dispense the heat generated. #### 4. Monitoring Temperature While adding acid, observe the temperature rise using a thermometer. The reaction is exothermic, so the temperature will increase. #### 5. Allow solution to cool Once the acid has been fully added, allow the solution to cool down to room temperature. #### 6. Labeling Properly label the prepared dilute sulfuric acid for future use. #### Observation A noticeable rise in temperature will be observed as the acid is added to water. No fumes or gases are emitted if the procedure is performed properly. #### Result Dilute sulfuric acid is successfully prepared. The temperature rise during the dilution process indicates the exothermic nature of the dilution process. #### Conclusion Concentrated sulfuric acid must be added to water slowly to prevent accidents due to its highly exothermic reaction. Proper safety precautions are essential when handling sulfuric acid to avoid injury or damage. The experiment was successful in producing dilute sulfuric acid. ## Experiment/Activity No. 4 ### Aim To prepare a standard solution of sodium hydroxide (NaOH) with a specific molarity and use it for titration purposes. ### Requirements - Sodium hydroxide (NaOH) pellets - Distilled water - Volumetric flasks (100mL or 250mL, depending on the desired volume of solution) - Funnel - Beaker (50mL or 100mL, depending on the desired volume of solution) - Weighing balance - Glass stirring rod - Pipette and pipette filler to dropper ### Procedure #### 1. Calculation Calculate the required mass of NaoH needed for the desired molarity (M) and volume (V) of the solution using the formula: ``` Mass of NaOH (g) = M \times Molar mass of NaOH (40g/mol) \times V (L) ``` For example: To prepare 100mL of 0.1M NaOH solution, the required mass of NaOH would be: ``` Mass of NaOH (g) = 0.1 \times 40/mol \times 0.1L ``` Therefore, the required mass of NaOH would be 0.4g. #### 2. Weighing Weigh the calculated mass of NaOH pellets on a weighing balance. #### 3. Dissolving NaOH Transfer the NaOH pellets into a clean beaker. Add about 50mL of distilled water to the beaker. Stir the solution using a glass stirring rod until the NaOH dissolves completely. Be careful, as the dissolution of NaOH is exothermic and may release heat. #### 4. Transfer to Volumetric Flask Once the NaOH solution has fully dissolved, transfer the solution to a volumetric flask using a funnel. Rinse the beaker and funnel with distilled water to ensure all the NaOH solution is transferred to the flask. #### 5. Dilution Add distilled water to the volumetric flask until the bottom of the meniscus touches the calibration mark. Stopper the flask and shake gently to ensure thorough mixing. #### 6. Label Clearly label the flask with the molarity and the date of preparation. #### Result A standard NaOH solution of the desired molarity (e.g. 0.1M) is prepared and ready for use in titration or other analytical experiments. #### Conclusion The preparation of NaOH solution involves careful weighing of the required amount of NaOH, dissolving it in distilled water, and proper dilution to the desired concentration. The solution can be used for various quantitative experiments such as acid-base titrations. ## Experiment/Activity No. 5 ### Aim To demonstrate the principle, working and maintenance of a pH Meter. ### Principle of a pH Meter A pH meter works on the principle of electrochemical measurement using a combination electrode. The electrode measures the voltage (or potential difference) generated between a glass electrode and a reference electrode when they are immersed in a solution. This potential difference is directly related to the hydrogen ion concentration (H⁺ ions) in the solution, which determines the pH. The relationship between the potential difference and pH is governed by the Nernst equation: ``` E = E<sup>o</sup> - (0.0591/n) * log[H<sup>+</sup>} ``` where: - E = Measured potential (in volts) - E^o = Standard electrode potential - n = Number of electrons exchanged - [H⁺] = Hydrogen ion concentration. ### Working of a pH Meter - **Calibration:** Before measurement, the pH meter must be calibrated using standard buffer solutions (usually of pH 4, pH 7 and pH 10). - **Measurement:** The glass electrode is sensitive to hydrogen ions and interacts with them in the solution. It develops a potential relative to the ion concentration. The reference electrode (usually filled with a solution of known pH) provides a stable reference voltage. The meter measures the potential difference between these two electrodes. This voltage is converted to a pH value by the pH meter circuitry. - **Display:** The meter displays the pH value, which ranges from 0 (very acidic) to 14 (very basic), with 7 being neutral. ### Maintenance of a pH Meter To ensure accurate and reliable results, regular maintenance of the pH meter is crucial: - **Cleaning:** After each use, rinse the electrode with distilled water to remove any residual sample or buffer solution. For stubborn deposits, clean the electrode by soaking it in a cleaning solution recommended by the manufacturer (e.g. diluted hydrochloric acid or pepsin for protein residues). - **Proper Storage:** Always store the electrode in a suitable storage solution (often a 3M KCl solution). Never store it dry, as this can damage the glass membrane and reference electrode. - **Calibration:** Regularly calibrate the meter using fresh buffer solutions. Calibrate at least once a day if you're using the meter frequently. - **Electrode Inspection:** Periodically inspect the electrode for any signs of damage, such as cracks in the glass bulb or depletion of the reference solution. Replace the electrode if necessary. - **Check for Response Time:** If the pH meter shows slow or unstable readings, it might be time to clean or replace the electrode. - **Avoid Contamination:** Always use fresh solutions and avoid cross-contamination between buffers or samples. Clean the electrode thoroughly between measurements. By adhering to these maintenance practices, the accuracy and lifespan of the pH meter will be prolonged. ## Experiment/Activity No. 6 ### Aim To demonstrate the proper techniques for cleaning glassware. ### Materials - Dirty glassware (e.g. beakers, flasks, test tubes) - Distilled water - Detergent solution or soap - Acetone or ethanol - Chromic acid / sulfuric acid mixture (for stubborn stains) - Soft bristled brush - Clean lint free cloths - Gloves and lab coat ### Procedure #### Step 1: Removal of Gross Contamination 1. Wear gloves and lab coat. 2. Remove large debris from glassware. 3. Rinse with distilled water. #### Step 2: Soaking 1. Fill sink or large container with detergent solution. 2. Soak glassware for 30 minutes to 1 hour. #### Step 3: Scrubbing 1. Use a soft bristled brush to scrub away stubborn stains. #### Step 4: Rinsing 1. Rinse thoroughly with distilled water. 2. Repeat rinsing process 2-3 times. #### Step 5: Drying 1. Dry glassware with clean lint free cloths. 2. Inspect for remaining streaks or residue. #### Step 6: Final Cleaning 1. Use acetone or ethanol to remove remaining residue. 2. Use chromic acid / sulfuric acid mixture for stubborn stains. #### Safety Precautions 1. Wear gloves and lab coat. 2. Avoid using abrasive materials. 3. Handle glassware carefully to avoid breakage. #### Result - Clean and sparkling glassware. - Free of streaks and residue. - Ready for use in laboratory experiments.