PRACTICE SCHOOL MODULE Submitted to &f/ ons! aS RAJIV GANDHI PROUDYOGIKI VISHWAVIDYALAYA BHOPAL (M.P.) In Partial Fulfillment of the Requirements for the Degree of Bachelor in Pharmacy (B. Pharm) Submitted By TANISHQ CHOUHAN Enrollment No:- 0847PY211092 LNCT SCHOOL OF PHARMACY VILLAGE KANADIYA, INDORE (M.P.) (Approved by AICTE, PCI New Delhi) (Affiliated to RGPV, Bhopal)LNCT SCHIOOL OF PHARMACY VILLAGE KANADIYA, INDORE (M.P.) (Approved by AICTE, PCI New Delhi) (Affiliated to RGPV, Bhopal) Certificate : This is to certify that Mr./Ms. TANISHQ CHOUHAN Enrollment No.847PY211092 In Partial Fulfillment of the Requirements for the Degree of Bachelor in Pharmacy (B. Pharm) by Rajiv Gandhi Proudyogiki Vishwavidyalaya, Bhopal had satisfactory completed 150 hr. training in ZEW/TH DRUGS LIMITED, DEPALPUR, INDORE, from 16-10-2024 to 30-10-2024 in academic session 2024-25. Place: - Indore Mr. Jinendra Sardiya Mr. Amit Joshi Incharge Principal Practice School Module‘(Formerty- Zenith Drage Private Limited) CONTACT Us: ZENIT! 2 3 © ntogrennnarugs com \ vovoxzas 4so74 CIN: U24234MP2000PLCON4ANS (g) Plant Ada. 7295 & 721,724, TUN, 7811, Murer (Orangpura, ‘Dhar Road. Near Kari, MOORE 453081 (MP) SYDMA Registered Office Address - K. Mo, 72/5, Village Muradaura, Depalpur, Indore, Madhya Pradesh - 433008 ( O ) ZENITH DRUGS LIMITED CERTIFICATE OF COMPLETION Mr.Tanishq chouhan Student of L.N.C.T. School of Pharmacy College, Kanadiya (Indore) has completed Internship during 16/10/2024 to 30/10/2024. We found him sincere, hardworking, and dedicated and result oriented. He worked well as part of the team during his tenure. We take this opportunity to thank him and wish all the best for his future. Date: 30/10/2024@ Acknowledgment This project consumed amount of work, research, & dedication. Still, implementation would not have been possible if I did not have a support of many individuals & organizations. Therefore I would like to extend our sincere gratitude to all of them. It is pleasure to express my deep sense of gratitude of thankfulness to Dr. Amit Joshi Sir, principle, LNCT School of Pharmacy, Indore. For his valuable guidance felicitous advice during the course of my practice school training practical. I wish to express my deep sense of gratitude to my incharge Mr. Jinendra Sardiya, Associated Professor, LNCT School of Pharmacy, Indore for her cooperation & valuable guidance throughout my B. Pharma practice school training practical. I am cordially grateful to my beloved parents, my family members & my friends who always covered their shade of love & blessing & provide their valuable moral support directly spirit & corporation. In order to widen my knowledge, to have new experiences in the field of health care, did training ZENITH DRUGS Pvt.Ltd. This training course is extended over a period of 150 hours, beginning on 16 october 2024 and ending on 30 october 2024. THANKS AGAIN TO ALL WHO HELPED ME. By TANISHQ CHOUHANPRACTICE SCHOOL ON PHARMACEUTICAL ANALYSIS (150 HOURS) ‘COURSE MODULES ‘TOPICS DURATION (RS) Module-1 Introduction to Pharmaceutical Analysis General introduction of analysis in Pharmacy. ‘Types of analysis in pharmacy. Different techniques of analysis. Importance of analysis in pharma industry.lmportance of Pharmacopeia in analysis Introduction to Standard operating procedures in analysis 1 Study of SOP 2.Preparation of SOP 3.lmportance of SOP Preparation of Reagents and Standard solutions Importance of reagents and standard solutions in analysis. 20hrs ‘Module-2 Theory, Principle, Instrumentation, Application of —_ following —_ Analytical Instruments 1 Analyticla balance 2.pH meter 3.UV Visible spectroscopy 4. IR spectroscopy 5.Fluorimetry 6.High performance liquid chromatography 7.HPTLC 8,.DSC 9.Dissolution 10.Particle size analyzer 11.NMR 12.Mass spectroscopy 12. Analytical method validation. ‘25hrs‘Module-3 Operation and Handling of Analytical Instruments 1, Analytical balance 2. pH meter 3. UV Visible spectroscopy 4. IR spectroscopy 5. Fluorimetry 6. High performance liquid chromatography 7.HPTLC 8.DSC 9.Dissolution 10.Particle size analyzer 4Shre ‘Module-4 Preparation of Analytical Methods For Various Drugs 1. Selection of drugs 2. Collection of Monographs 3. Literature Search 4, Selection of equipment 5. Method Development 6. Method Validation 7. Quantification of Drugs, 25hrs ‘Module-S Calibration of Analytical Inetruments 1. Definition and importance of calibration in pharmacy. 2. Calibration of Glassware 3. Calibration of following instruments: 1. Analytical balance 2. pH meter 3. UV Visible spectroscopy 4. IR spectroscopy 5. Fluorimetry 6. High performance liquid chromatography 7. HPTLC 8. DSC 25hrs Module-6 Good Documentation Practice 1. Writing of Laboratory Note Book 2. Preparation of Protocols 3. Preparation of Validation Reports 10hrs7 ore Figure Titles vase Figure 1 | Analytical balance 12 Figure2 | Ph Meter 4 Figure 3 | UV-Visible Spectroscopy Is Figure 4 | IR Spectroscopy 7 Figure 5 | Fluorimetry 18 Figure 6 | High-Performance Liquid Chromatography(HPLC) 20 Figure 7 | High-Performance Thin Liquid Chromatography(HPTLC) 2 Figure 8 | Differential Scanning calorimetry (DSC) 2B Figure 9 | Dissolution Tester 4 Figure 10 | Particle Size Analyzer 25 Figure 11 | Nuclear Magnetic Resonance (NMR) Spectroscopy 26 Figure 12 | Mass Spectroscopy 27MODULE - 01 + Introduction to pharmaceutical Analysis General Introduction - The pharmaceutical analysis is a branch of chemistry, which involves the series of process for the identification, determination, quantitation, and purification, @ Types of analysis in pharmacy : 1. Chromatography - A common technique that separates and identifies species in organic, inorganic, and biological materials. High performance liquid chromatography (HPLC) is a type of chromatography that is often used in the pharmaceutical industry. 2. Spectroscopic techniques - A powerful and cost-effective tool for the qualitative and quantitative analysis of pharmaceutical and_ biological materials 3. Mass Spectrometry - A sensitive and reproducible technique that separates complex pharmacological mixtures into their components a Titrimetric techniques - A classical method that determines the volume of a sample after a chemical reaction has occurred. 5. Electrophoresis - A versatile technique that separates analytes within a capillary based on their electrophoretic mobilities. @ Importance of Pharmacopeia in analysis : 1. Quality standards - Pharmacopoeias define the quality standards for medicines and the substances used to make them, including purity criteria for raw materials and containers,mown Provide analysis methods - Pharmacopoeias include methods for quality control analysis. . Help ensure quality - Pharmacopoeias help ensure that medicines are reliable, effective, and safe for patients. |. Help make medicines available - Pharmacopoeias help ensure that medicines are widely available to patients around the world . Help reduce risk of side effects - Pharmacopocias are updated to reflect changes in the drug market, which can help reduce the risk of unwanted side effects. Introduction to Standard operating procedures in analysis : Study of SOP (Standard Operating Procedure) Preparation of SOP (Standard Operating Procedure) Importance of SOP (Standard Operating Procedure) . Study of SOP (Standard Operating Procedure) - A study of standard operating procedures (SOPs) may evaluate the effectiveness of SOPs in improving the quality and safety of a process or procedure. For example, a study may evaluate the clinical effectiveness of integrating SOPs with teamwork training (TT) to improve the quality and safety of surgery. Preparation of SOP (Standard Operating Procedure) - ¥ Determine the scope and format: Decide what the SOP will cover and how it will be presented. v Identify the stakeholders and creators: Determine who will be responsible for creating and using the SOP. ¥ Gather information: Collect all the information needed to create the SOP. Y Write the SOP: Include the following in the SOP: 93. 1. A title page and table of contents 2. The specific procedures 3. Standards, regulatory requirements, roles and responsibilities, and inputs and outputs 4. Health and safety warnings 5. A troubleshooting section 6. A complete list of all equipment and supplies ¥ Review and validate: Review the SOP and validate it. ¥ Implement and train: Implement the SOP and train the end-users. ¥ Regular review and updates: Review and update the SOP regularly Importance of SOP (Standard Operating Procedure) : Y Consistency - SOPs ensure that everyone in the organization follows the same process, which leads to consistent results and quality assurance. Y Safety - SOPs outline safe practices and procedures to reduce the risk of accidents and injuries. Y Efficiency - SOPs streamline processes and reduce the time it takes to complete routine tasks Y Training - SOPs can simplify employee training, saving time and money. ¥ Communication - SOPs can help streamline communication by reducing the chance of miscommunication. Y Regulatory compliance - Some industries, like health care, food, and manufacturing, require SOPs for compliance purposes. V Knowledge preservation - SOPs document best practices for future reference Y Accountability - Digital SOP assignments can provide accountability for business activities.MODULE - 02 * Principle, Instrumentation, Application of following Analytical Instruments : 1. Analytical balance - Analytical balances are an extremely accurate laboratory balance created to precisely measure the mass of an object > Principles - Analytical balances work on the principle of magnetic force restoration (MFR) to determine the weight of an object. i. Detect the sample's force- When an object is placed on the balance pan, it exerts a downward force. This force is detected by sensors. ii.Generate an opposing force - An electromagnet generates an opposingforce that balances the weight of the sample iii, Measure the force - The current required to maintain the balance is proportional to the object's mass. This current is measured to determine the mass iv. Display the mass - The mass is displayed on the balance screen Analytical balances are often housed in a protective draft shield to isolate the weighing chamber from external factors like air currents and vibrations. To ensure accurate weighing, you can: Raise the humii ty level to 40% Conduct weighing operations while standing on anti-static flooring Avoid storing samples in plastic containers Maintain a constant temperature in the environment and for the weighing equipmentAnalytical Balance Glass Door Balance Pan Door Handle Anti-draft Ring Display Tare Button ‘Mode Button Level Adjustment Feet. Power Button > Application of Analytical Instruments - Analytical balances, also known as "lab balances", are precision measuring instruments used in a variety of industries to determine the mass of solid objects, liquids, powders, and granular substances: i. Sample preparation - Analytical balances are used to prepare samples for analysis in laboratories and research centers. ii, Formulation and recipe calculation - Analytical balances are used to calculate recipes and formulations. iii, Density determination - Analytical balances are used to determine the density of substances. iv. Quality control testing - Analytical balances are used to ensure the quality of products by performing quality control testing, y. Environmental and pollution level analysis - Analytical balances are used to analyze the level of pollution in water, soil, or air.vi. Food industry - Analytical balances are used to ensure the quality of food products by accurately weighing ingredients. vii. Pharmaceutical industry - Analytical balances are used in the pharmaceutical industry. Plastics and chemical manufacturing - Analytical balances are used in plastics and chemical manufacturing. 2, pH meter - A pH meter is a scientific instrument that measures the hydrogen- ion activity in water-based solutions, indicating its acidity or alkalinity expressed as pH pH meter is an instrument used to measure acidity or alkalinity of a solution - also know as pH. pH is the unit of measure that describes the degree of acidity or alkalinity. It is measured on a scale of 0 to 14. > Priciples - The working principle behind pH meters is potentiometry. This is the measurement of a solution's electric potential (voltage). Remember how acidic solutions can efficiently conduct an electric current because of the positive hydrogen ions? The ability of a solution to conduct a current is called electric potential, > Application of pH meter - The pH is defined as a measure of acidity or alkalinity of a solution, which is why its control is crucial in the pharmaceutical world. The pH can significantly influence various characteristics of: Drugs Such as their stability Efficacy Solubility Ultimately their Bioavailability. S& S443. UV Visible Spectroscopy - Ultraviolet-visible (UV-Vis) spectroscopy is a technique that measures how light in the UV and visible regions of the electromagnetic spectrum is absorbed, transmitted, or reflected by a sample. > Principles - The principles of UV-visible spectroscopy are based on the interaction between light and matter, and include: i, Absorption - When a chemical compound absorbs light, the electrons in the compound are excited and jump from a ground state to an excited state. The difference in energy between the two states is equal to the amount of light absorbed. ii, Beer-Lambert-Bouguer law - This law is the basis of UV-visible spectroscopy iii, Absorption maximum - The wavelength of light that a molecule absorbs most strongly is called the absorption maximum. iv. Spectrophotometer - A Spectrophotometer measures the amount of absorption at different wavelengths. y. Monochromator - A Monochromator is a disk with slits that can be adjusted to select a specific wavelength. A double Monochromator is used in high-performance instruments to reduce stray light and increase spectral accuracy.vi. Light source - A steady light source that emits a wide range of wavelengths is required. A xenon lamp is commonly used, but it is more expensive and less stable than tungsten or halogen lamps. tion of UV Visible Spectroscopy - UV-Vis spectroscopy is a common analytical technique used in the pharmaceutical industry for a variety of applications, including: i. Drug discovery and development - UV-Vis spectroscopy is used to develop active pharmaceutical ingredients (APIs) ii. Quantifying impurities - UV-Vis spectroscopy is used to quantify impurities in drug ingredients and products. iii, Dissolution testing- UV-Vis spectroscopy is used to analyze the results of dissolution testing for solid oral dosage forms like tablets.iv. Chemical identification and quantification - UV-Vis spectroscopy can confirm the chemical identity and quantify the purity of drugs and drug ingredients. v. Structure elucidation - UV-Vis spectroscopy can be used to identify characteristic absorption bands to elucidate the structure of organic compounds. vi. Chemical kinetics - UV-Vis spectroscopy can be used to monitor reaction rates and determine acid/base dissociation constants. IR spectroscopy - Infrared (IR) spectroscopy is a chemical analysis technique that measures the interaction of infrared light with matter to identify chemical substances and functional groups. > Principles - The principle of infrared (IR) spectroscopy is that molecules absorb specific frequencies of light that are characteristic of their structure: i, Light absorption - A molecule absorbs light when its structure has a bond with a dipole moment, meaning that the electrons within the bond are not shared equally. ii. Vibrational frequencies -The absorbed radiation matches the vibrational frequency of the molecule. This is called a resonant frequency. iii, Energy - The energy of the absorbed light is affected by the shape of the molecule's potential energy surfaces, the masses of the atoms, and the associated vibronic coupling. iv. Vibrations - The vibrations that can be induced by infrared light are limited to those involving a change in dipole moment. These vibrations can be stretching, bending, or rotational. > Application of IR spectroscopy - Infrared (IR) spectroscopy has many applications, including: i, Chemical identification - IR spectroscopy is used to identify unknown samples, especially in forensics, damage analysis, and competition analysis. ii. Quality control - IR spectroscopy is used to ensure the integrity of chemical products and to ensure quality control 16iii. Forensic analysis - [R spectroscopy is used in both criminal and civil cases. For example, it can be used to identify polymer degradation or determine the blood alcohol content of a suspected drunk driver. iv. Material verificatio. IR spectroscopy can be used for material verification. y. Quantification of components - IR spectroscopy can be used to quantify components. vi. Micro chemical analysis - IR spectroscopy can be used for micro chemical analysis. vii. Chemical imaging - IR spectroscopy can be used for chemical imaging Catalysis research - IR spectroscopy can be used to characterize catalysts, detect intermediates, and detect products during catalytic reactions. ix. Dynamic measurement - IR spectroscopy can be used for dynamic measurement. x. Monitoring applications: IR spectroscopy can be used for monitoring applications, such as measuring CO2 concentrations in greenhouses and growth chambers. xi. Polymer manufacture: IR spectroscopy can be used to measure the degree of polymerization in polymer manufacture.5. Fluorimetry - Fluorimetry, also known as fluorescence spectroscopy or spectrofluorometry, is a technique that measures fluorescence in a sample to analyze molecular parameters of biological, biochemical, and chemical processes. > Principlers - The principle of fluorimetry is the measurement of fluorescence, which is the visible light emitted by a substance after it absorbs light of a specific wavelength i, Light absorption ii, Electron relaxation iii, Light emission> Appliction of Fluorimetry - Fluorometry is ordinarily more sensitive than absorbance measurements. It is a widely accepted and powerful technique that is used for a variety of environmental, industrial, medical diagnostics, DNA sequencing, forensics, genetic analysis, and biotechnology applications. . High performance liquid chromatography - High Performance Liquid Chromatography (HPLC) is an analytical technique that is ubiquitous in labs across the globe. This technique is commonly used in labs for the purpose of separating, identifying, and quantifying components in a mixture. > Principles of High Performance Liquid Chromatography (HPLC) - The principles of High Performance Liquid Chromatography (HPLC) include i. Separation ii, Absorption iii, Mobile phase iv. Retention time v. Detectors a . Efficiency > Application of High Performance Liquid Chromatography (HPLC) - High Performance Liquid Chromatography (HPLC) is a technique used to identify, quantify, and purify compounds and analytes. It has many applications, including: i, Pharmaceutical i. Food iii. Water purification iv. Forencis v. Environmental vi. Chemical Sciencevii. Bloptechnology viii Chemical Separation ix. Research x. Manufacturing xi. Bio-monitoring of pollution7. High-Performance Thin Layer Chromatography (HPTLC) - High- performance thin-layer chromatography (HPTLC) is a technique that uses thin- layer chromatography (TLC) to analyze compounds and mixtures. It's a versatile, cost-effective, and efficient method that's used in many fields, including pharmaceuticals, food and drug analysis, and environmental analysis. » Principles - The components with larger efficiency for stationary phase interact with the surface and move slowly in the mobile phase whereas components with lesser affinity for stationary phase move fast. This variation in the movement of components results in the separation of components on the chromatographic plate > Application - i, Water purification. ii, Detection of impurities in pharmaceutical industries. iti. Pre-concentration of trace components. iv. Ligand-exchange chromatography. v. Ion-exchange chromatography of proteins. vi High-pH anion-exchange chromatography of _ carbohydrates oligosaccharides and8. Differential Scanning Calorimetry (DSC) - Differential scanning calorimetry (DSC) is a technique that measures the difference in heat required to raise the temperature of a sample and a reference material. The sample and reference are kept at nearly the same temperature throughout the experiment. > Principles - The principle of differential scanning calorimetry (DSC) is that the amount of heat required to maintain a sample's temperature changes when the sample undergoes a physical transformation. This is because the sample absorbs or releases heat during the transformation, which is called an exothermic or endothermic process. > Application - Differential Scanning Calorimetry (DSC) is a highly sensitive technique to study the thermotropic properties of many different biological macromolecules and extracts. Since its early development, DSC has been applied to the pharm +aceutical field with excipient studies and DNA drugs.9. Dissolution - A dissolution machine, also known as a dissolution test apparatus, is a laboratory device that measures how quickly and to what extent a drug is released from a dosage form into a solution. Dissolution machines are used in pharmaceutical development and quality control to ensure consistent drug quality and efficacy. > Principles - The principle of dissolution is that a solute will dissolve in a solvent when the solute and solvent have the same polarity. This is known as the "like dissolves like" principle. For example, salt dissolves in water because both are polar compounds, but salt does not dissolve in oil because oil is nonpolar. > Application - In the pharmaceutical industry, dissolution is the process of a solid drug entering a solution, and is a key quality control test for solid and semi-solid dosage forms. Dissolution is important because drugs must be in solution to be absorbed and have an effect on the body. 2310. Particle size analyzer - A particle size analyzer (PSA) is a device that measures the size of particles in a sample by analyzing how light interacts with them. PSAs are used to determine the size distribution of powders, suspensions, or emulsions. Here are some types of particle size analyzers. > Principles - A particle size analyzer works on the principle that when a beam of light is scattered by a group of particles, the angle and intensity of the scattered light is inversely proportional to the particle size. > Application - Particle size is a crucial parameter in the pharmaceutical industry, because it influences surface area and porosity and, hence, has an impact on bioavailability, effectiveness and shelf life of a drug.11, Nuclear Magnetic Resonance Spectroscopy (NMR) - Nuclear magnetic resonance (NMR) spectroscopy is a non-destructive analytical technique that uses the magnetic properties of atomic nuclei to study the physical, chemical, and biological properties of matter. > Principles - Working principle of nuclear magnetic resonance (NMR) is based on the spins of atomic nuclei. Nuclei with an odd mass or odd atomic number have “nuclear spin" (in a similar fashion to the spin of electrons). Since a nucleus is a charged particle in motion, it will develop a magnetic field. > Application - NMR spectroscopy is one of the principal techniques used to obtain physical, chemical, electronic and structural information about molecules due to the chemical shift of the resonance frequencies of the 25nuclear spins in the sample. Peak splittings due to J- or dipolar couplings between nuclei are also useful ‘Computer Console Super-conducting magnet Instrument control and Transmit and receives the Generates a powerful magnetic data processing radio-requency waves used to || field that is tens of thousands make the NMR measurements. ‘of times stronger than the earthis magnetic field, Samples are placed within this magnetic field and exposed to fadio waves. 12, Mass spectroscopy - Mass spectrometry is an analytical tool useful for measuring the mass-to-charge ratio (m/z) of one or more molecules present in a sample. These measurements can often be used to calculate the exact molecular weight of the sample components as well. > Principles - Mass spectrometry (MS) is an analytical technique that separates ionized particles such as atoms, molecules, and clusters by using differences in the ratios of their charges to their respective masses (mass/charge; m/z), and can be used to determine the molecular weight of the particles.> Application - Mass spectrometry (MS) is a powerful qualitative and quantitative analytical technique used to identify and quantify a wide range of clinically relevant analytes. When coupled with gas or liquid chromatographs, mass spectrometers allow the expansion of analytical capabilities to various clinical applications. 13. Analytical method validation - Analytical method validation is a critical process in the pharmaceutical, biotechnology, and food industries to ensure the quality and safety of products. The objective of the validation of an analytical method is to demonstrate that it is suitable for its intended purpose. > Principles - The three main validation principles in product design and construction are quality, safety, and efficacy. Inspection of the finished product and work in progress alone cannot ensure quality. To ensure that the final product meets all quality requirements, every step of the production process is examined.MODULE - 03 > Operation and Handling of Analytical Instrument : 1. Analytical balance - Analytical balances measure the weight of a sample by using an electromagnet to generate a force that counteracts the sample's mass. > Operation - i. Place the sample - Put the sample on the balance's weighing pan ii. Detect the force - The sample's mass creates a downward force that is detected by sensors. iii. Generate a counterforce - The sensors activate an electromagnet that generates a force to counter the sample's weight. iv. Measure the force - The balance measures the force required to maintain the balance and displays the weight Analytical balances are often used in laboratories for research and development, quality control, and other purposes. They can be very accurate, often measuring down to the microgram level Here are some things to keep in mind when using an analytical balance: v Environmental factors - Air currents, vibrations, temperature, and gravitational acceleration can all affect the balance's measurements. To reduce the impact of these factors, you can use a draft shield and maintain a consistent temperature. Y Calibration - The balance may need to be calibrated to account for gravitational differences between locations and altitudes. Cleaning - Clean the weighing pan before and after each use to prevent contamination, ¥ Object placement: Use tweezers to place objects on the balance. > Handling -i. Temperature - Allow the balance to warm up and reach the correct operating temperature, which can take at least two hours. ii. Location - Place the balance on a firm, level surface, such as a built-in countertop or a heavy table. Avoid areas with drafts, vibrations, or air conditioning systems. Calibration - Compare the balance's measurements to a known standard or reference weight iv. Weighing - Close the balance doors to prevent air currents from affecting the reading, Use a stopper container to weigh the sample, and don't handle it with bare hands. Place the object to be weighed on the left pan and the weights on the right. v. Cleaning - Clean up spills immediately with a cloth dampened with mild soap suds. Don't use aggressive cleaning agents or solvents. =. i, Storage - Store the balance in a clean, dry area with controlled temperature and humidity levels. Protect it from direct sunlight, excessive heat, and chemicals that may corrode it vii. Accessories - Use an electrostatic discharge ionizer to improve accuracy when weighing non-conductive materials like glass, china, and plastic. viii. Servicing - Only trained technicians authorized by the manufacturer should service the balance. 2. pH meter - A pH meter is a scientific instrument that measures the hydrogen- ion activity in water-based solutions, indicating its acidity or alkalinity expressed as pH. > Operation - i. Electrodes - A pH meter has two electrodes: a reference electrode and a sensor electrode. The reference electrode contains a substance with a known electric potential, and the sensor electrode is inserted into the solution being tested ii, Glass membran - The sensor electrode has a glass membrane with a buffer solution that allows hydrogen ions to enter the membrane. 29iii. Electrical circuit - When the electrodes are inserted into the solution, an electrical circuit is completed and a potential difference is created iv. Voltage measurement - The pH meter measures the voltage potential and converts it to a pH value. v. Calibration - Before each use, a pH meter is calibrated with solutions of known pH to ensure accuracy. Here are some tips for using a pH meter: Y Rinse the electrodes with deionized water before and after dipping them into the sample solution. v Ensure the pH electrode bulb is immersed in the solution, v Read the pH value once it has stabilized. Clean the electrodes after use by rinsing with deionized water and placing the electrode in pH4 solution. v If instructed, place the pH electrode in the pH storage solution. > Handling - i. Calibration - Calibrate the pH meter before each use to ensure accurate readings. ii, Electrode care - Rinse the electrode with deionized or distilled water before and after each measurement. Don't wipe the electrode with a cloth or towel, as this can cause static and affect readings. Store the electrode in its storage solution when not in use. Solution handling - Handle buffer solutions with care. Wear gloves, goggles, and lab coats when handling solutions. iv. Sample handling - Use separate containers for different samples to prevent cross-contamination. y. Electrode immersion- Gently immerse the electrode into the solution. Don't force it in.vi. Rinsing - Rinse between measurements to prevent contamination. vii.Stirring - Stir the pH buffers and sample at the same rate to get a representative pH value. viii, | Temperature - Use a 3-in-1 pH electrode or a combination pH electrode and temperature probe to compensate for temperature effects on pH. ix. Protein deposits - Soak in 1% pepsin in 0.1 M HCI for 5 minutes. 3. UV Visible spectroscopy - Ultraviolet-visible (UV-Vis) spectroscopy is an analytical technique that measures how much ultraviolet or visible light a sample absorbs or transmits. It's a type of electron spectroscopy that uses photons in the ultraviolet (100-400 nm), visible (400-750 nm), or near infrared (750-1400 nm) range. > Operation - i. Illuminate the sample - A UV-Vis spectrophotometer shines light across the UV to visible wavelength range (typically 190 to 900 nm) onto a sample ii, Meaeasure the light - The spectrophotometer measures how much light the sample absorbs, transmits, or reflects at each wavelength. iii. Calculate absorbance - The transmittance (T) is used to calculate the absorbance (A) using the formula A= -log (T) iv. Analyze the spectrum - The resulting absorbance spectrum shows how a compound absorbs light at different wavelengths. Here are some things to consider when using a UV-Vis spectrophotometer : v Prepare a serial dilution - Create a series of standard solutions with concentrations that are evenly spaced. Y Use a blank - Place a blank in both the measurement and reference beam lines, then run a background.4. v Save your data - Save your data as a csv file, and make sure to save the results to a shared folder. v Clean the cuvette -After each sample, clean the cuvette with compressed air and empty it into the organic waste. > Handling - i, Turn on and warm up - Turn on the spectrometer and let the lamps warm up for about 20 minutes to stabilize them. ii, Prepare the cuvette - Clean the outside of the cuvette and fill it with a solvent to act as a blank. iii. Align the cuvette - Make sure the cuvette is properly aligned in the spectrometer. Some cuvettes have two sides, one for handling and one for light to pass through. iv. Take a blank reading - Take a reading for the blank, and subtract any absorbance from future samples. v. Prepare the sample - For most samples, dissolve them in a solvent that won't produce its own results. The solution should be concentrated enough to allow light to pass through. i, Pipette the sample - Use a pipette to add the sample to the cuvette vii. Insert the cuvette - Insert the cuvette into the spectrometer and close the lid. viii, Avoid touching the cuvette - Don't touch the part of the cuvette that light passes through, as fingerprints can interfere with the absorbance reading ix, Use a cuvette holder - Use a cuvette holder when injecting the sample. IR spectroscopy - Infrared (IR) spectroscopy is a technique used in the pharmaceutical industry to identify and study pharmaceutical substances, and to analyze samples for contaminants. High performance liquid chromatography - High Performance Liquid Chromatography (HPLC) is an analytical technique that is ubiquitous in labs across the globe. This technique is commonly used in labs for the purpose of separating, identifying, and quantifying components in a mixture.> Operation - i, Sample injection - A liquid sample is injected into a solvent stream called the mobile phase. ii. Column separation - The sample passes through a column packed with a separation medium called the stationary phase. The sample components separate as they move through the column at different velocities. iii. Detection - The sample components emerge from the column and are detected by one or more detectors. The detector measures the components and sends a voltage response to a chromatography data system (CDS). iv. Data analysis - The CDS translates the detected signal into a chromatogram, which is a graph that shows the separation of the sample components. The x-axis of the chromatogram represents time, and the y- axis represents the signal generated by the detector. The time at which a peak emerges identifies the sample component, and the area of the peak Tepresents the quantity. Here are some tips for using HPLC: Y Use a syringe that's the correct size to match the size of the loop. ¥ Rinse the syringe with the solvent used in the sample. Y Make sure there are no air bubbles in the syringe. Y Before running the actual samples, run a standard. Ensure the pressure is stable. > Handling - i, Clean the syringe - Use a syringe that matches the size of the loop, and rinse it with the solvent used in the sample. ii, Check the leve - Make sure the lever is in the correct position to receive the sample.Check for air bubbles - Ensure there are no air bubbles in the syringe. iv. Set the waste line - Confirm that the waste line is in a waste container and not recycling back into the mobile phase. v. Set the flow rate - Set the flow rate of the mobile phase to 0.5 mL/min. vi. Set the pressure - Set the minimum and maximum pressure on the solvent delivery system. vii. Set the blank - Press "zero" on the detector's front panel to set the blank. viii, Store the column - Store the HPLC column in a methanol-water system in a 70:30 ratio when not in use ix. Avoid mechanical damage - Avoid any mechanical damage to the HPLC. x. Avoid sudden fluctuations - Avoid sudden fluctuations in the mobile phase composition, temperature, and flow rate. High-Performance Thin Layer Chromatography (HPTLC) - High- performance thin-layer chromatography (HPTLC) is a technique that uses thin- layer chromatography (TLC) to analyze compounds and mixtures. It's a versatile, cost-effective, and efficient method that's used in many fields, including pharmaceuticals, food and drug analysis, and environmental analysis. > Operation - i. Sample application - Samples are applied to pre-coated plates with a thickness of 150-200 um and a particle size of 5-7 um. ii, Separation - The components of the sample are separated on the plate. 4i. Detection - The separated materials are detected. iv. Data collection - Data is collected from the separated materials. v. Quantitative analysis - A scanner is used to determine the amount of each compound in the sample. A graph is then created and compared to standards to determine the concentration of the compound. HPTLC offers several advantages over traditional TLC, including: ¥ Improved resolution - HPTLC uses higher-quality plates with finer particles, which improves resolution. ¥ Lower limit of detection - HPTLC has a lower limit of detection (LOD) than traditional TLC. ¥ Automation - HPTLC can use automatic or semi-automatic sample application, and can automatically dry the plate Y Less exposure to toxic solvents - HPTLC can reduce exposure to toxic solvents ¥ Reduced environmental pollution - HPTLC can reduce environmental pollution. Handling - Handling the column - Be careful when handling the column, as it can be fragile. Use protective cases or column guards to transport or store the column. Flow rate - Maintain the recommended flow rate for the column. Exceeding the flow rate can cause the column to degrade, fail, or have reduced resolutioniii. Mobile phase - Store the mobile phase in borosilicate glass bottles with a narrow opening. Cover the opening to prevent evaporation and oxygen intake. If the mobile phase is photosensitive, store it in an amber bottle or cover it with aluminum foil. iv. Mixing solvents - Mix solvents to achieve a homogeneous mobile phase composition. This will ensure equal retention times for each injection. v. Column lifetime - Monitor the column's performance, such as peak shape, efficiency, and retention time stability. Establish a schedule for replacing the column based on these evaluations. vi. Documentation - Keep detailed records of the column's usage, including the serial number, installation date, column type, and usage history. vii. Injection - Use the correct syringe size to match the loop size. Rinse the syringe with the sample's solvent, then displace the solvent into a waste container. Inject the sample into the injector piece. viii. | Cleaning - Flush the column with a strong solvent at the end of the run to clean it Differential Scanning Calorimetry (DSC) - Differential scanning calorimetry (DSC) is a technique that measures the difference in heat required to raise the temperature of a sample and a reference material. The sample and reference are kept at nearly the same temperature throughout the experiment. > Operation - i. Prepare the sample - Use a small sample in a crucible made of an inert material like metal or ceramic. ii, Perform the experiment - Heat or cool the sample, or hold it at a constant temperature. iii, Collect the data - A sensor detects the energy absorbed or released by the sample, and the difference in heat flow between the sample and a reference is plotted.iv. Analyze the data - Use thermal analysis software to evaluate the shape of the curve, which shows endothermic or exothermic peaks for thermal events, DSC can be used to study a variety of materials, including: v v v v v v v Biopolymers like proteins and enzymes Metals, alloys, films, and fibers Oils and greases Amorphous substances Polymers Pharmaceuticals Powders DSC can help determine a material's thermal characteristics and behavior, including: KKK KKK Stability Phase transitions Melting behavior Chemical reactions Glass transition Specific heat capacity Purity > Handling - i. Sample preparation - The sample should be small and precisely measured, and placed in a sample crucible or pan, The sample should be encapsulated in 2 metal pan, usually aluminum, ii, Reference material - An empty crucible of the same type as the sample is used as @ reference to calibrate the system. Temperature program - Set the start and end temperatures, as well as the heating and cooling rates.iv. Furnace gas - Select the appropriate furnace gas depending on whether an inert or oxidizing atmosphere is required. v. Placing the crucibles - Place the sample and reference crucibles into the furnace, either manually or automatically. vi. Monitoring the heat flow - The DSC instrument detects the difference in heat flow between the sample and reference crucibles. vii. Plotting the results - The results are plotted on a measurement curve that shows the enthalpy changes of the sample. viii, Analyzing the results - The heat flow is analyzed to identify phase transitions and reactions, Dissolution - A dissolution machine, also known as a dissolution test apparatus, is a laboratory device that measures how quickly and to what extent a drug is released from a dosage form into a solution. Dissolution machines are used in pharmaceutical development and quality control to ensure consistent drug quality and efficacy. > Operation - i. Ensure the instrument is clean and free of dust. ii, Tur on the power and heater switches. ii, Set the sampling time interval and RPM. iv. Place the jars filled with the medium in the tank V. Attach the paddle to the spindle and tighten it. vi. Lower the stirrer unit Vii. Set the temperature and RPM parameters. Vii. Start the stirrer. ix. Once the desired temperature is reached, leave the tablets in the test jars or baskets. x. Start the test. xi, Perform the analytical procedure and calculate the results. xii, Tur off the instrument. > Handling - 38i. Dissolution medium - A liquid is placed in the vessels of a dissolution unit. The medium can be deionized water, chemically-prepared solutions, or mediums with surfactants. The medium should be degassed to remove dissolved gases, which can affect the results. ii, Drug placement - The drug is placed in the medium once it has reached the correct temperature Dissolution apparatus - The dissolution apparatus is operated. The apparatus can be manual or an instrument that automates the process. iv. Sample detection - Sample solutions are detected using HPLC and Ultra violet visible spectroscopy y. Acceptance criteria - The tested samples must meet the criteria of release specifications. © The dissolution procedure should be reproducible and robust enough to be transferred between laboratories. The acceptance criteria should be representative of multiple batches with the same composition and manufacturing process. © Dissolution is important for health practitioners because drugs must be in solution to be absorbed and have a physiological effect on the human body. 10. Particle size analyzer - Particle Size Analyzer is widely used method to characterize the sediment particles based on diffraction of a laser light source by the samples under analysis, The analyzer is used to determine the size distribution of a powder, a suspension or an emulsion, based on light diffraction > Operation - i. Laser diffraction - A monochromatic source, like a laser, illuminates the particles, creating a diffraction pattern. The variation in the intensity of the scattered light is measured, with smaller particles scattering at a smaller angle and larger particles scattering at a larger angle. ii, Image analysis - This can be static or dynamic. iii. Sieving - A traditional method for measuring particle size © Particle size analysis is a standard procedure in research and quality control labs to ensure the quality of the final product. It's important in many industries, including food, chemical, mining, agriculture, cosmetics, pharmaceuticals, energy, and aggregate, 39MODULE - 04 % Preparation of Analytical Methods For Various Drugs : 1, Selection of drugs - When selecting an analytical method for a drug, the chemical properties of the drug are the primary consideration, The following factors should be considered when selecting an analytical method: i Chemical properties - The drug's ionizability, polarity, and presence of functional groups (chromophores). Purpose and Scope - The specific purpose and scope of the analysis should determine the analytical instruments and methods used. Regulatory guidelines - Guidelines from the US Food and Drug. Administration (USFDA) and the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) should be followed. Some common methods for drug analysis include: ¥ Gravimetric analysis - A basic chemical method for drug analysis and testing. v Acid-base titration - A reagent solution of known concentration is added to the tested substance until the chemical reaction is complete. Y Spectroscopy - The drug is radiated through different frequencies. ¥ Chromatography - A method for seperating and analyzing compounds in a mixture v Electrophoresis - An important method for biochemical drug analysis and biotechnology 2. Collection of Monographs - Analytical methods for drugs are included in compendial monographs to characterize the quality of bulk drug materials. Some analytical techniques used in pharmaceutical analysis include i. Chromatography - A selective technique that can be used to identify particulates and residues, Some types of chromatography include: 40Y Thin-layer chromatography (TLC) - A technique that uses a reference substance, ¥ Infrared (IR) - A technique that is faster and more specific than TLC. v Liquid chromatography (LC) - A technique that can perform identity, purity, and assay in the same run. v Ton chromatography (IC) - A technique that can simultaneously determine multiple anions or cations in a single injection ii, Spectrometry - A technique that can be used to identify particulates and residues. Microscopy - A technique that can be used to identify particulates and residues. iv. Titrimetry - An assay method that can be used in compendial monographs. v. Capillary electrophoresis - An assay method that can be used in compendial monographs vi. Electro analytical methods - An assay method that can be used in compendial monographs © Method validation is a process that confirms that an analytical procedure is suitable for its intended use, The results of method validation can be used to judge the quality, reliability, and consistency of analytical results 3. Literature Search - A literature review is a key step in the process of developing analytical methods for drugs: i. Sources - Search for information in books, periodicals, chemical manufacturers, and regulatory agency compendia like USP/NF. You can also use the Chemical Abstracts Service (CAS) for automated literature searches ii, Topies - Look for information on the analyte's synthesis, physical and chemical properties, solubility, and relevant analytical methods. iii, Pharmaopeias - Check pharmacopeias like IP, USP, EP, BP, and JP for suitable analytical methods. iv. Chromatographic journals - Look for suitable analytical methods in chromatographic journals. aOther steps in the process of developing analytical methods for drugs include: Defining the analytical method's objectives, Developing a method plan, Optimizing the method, and Validating the method, ‘The validation process involves evaluating the method's accuracy, precision, specificity, linearity, range, LOD, LOQ, ruggedness, and robustness. Some common analytical techniques for drug analysis include: Spectroscopy Chromatography Electrochemical techniques Electrophoretic Flow injection analysis DNA amplification SS KKK KS Thin layer chromatography Selection of equipment - Here are some things to consider when preparing analytical methods for different equipment i. Sample preparation - This is one of the most important steps in the analytical process. The sample preparation method determines the accuracy and precision of the analytical method. ii, Calibration standard preparation - For almost all analytical methods, a series of calibration reference standard solutions with different concentrations are prepared. The results are plotted after putting the calibration solutions through the same analytical procedure as the sample Reference standards - Reference siandards are used to identify or determine the concentration of an analyte in a sample. The reference standard is prepared with a known concentration so that the unknown sample can be compared to it. iv, Reference materials - Certified Reference Materials (CRMs) are essential for analytical chemistry and clinical analysis. Most analytical instrumentation is comparative, so it requires a sample of known composition for accurate calibration y. Sampling - Correct sampling is a vital link in the preparation chain. All subsequent steps in the process rely on accurate sampling. 2MODULE - 05 “ Calibration of Analytical Instruments - 1. Definition and importance of calibration in pharmacy - Calibration in the pharmaceutical industry is the process of comparing a measurement device's results to a standard fo ensure the accuracy and safety of the equipment. It's a key part of quality assurance and manufacturing standards. Calibration is important for many reasons, including: i, Safety - Calibration ensures that the equipment produces safe and accurate results. ii, Quality standards - Calibration helps ensure that the quality of the product isn’t affected by constant errors Patient safety - Calibration ensures that experimental results are consistent and reproducible, which helps develop new drugs and therapies. iv. Cost efficiency - Calibration can reduce waste and increase operational efficiency. v. Compliance - Calibration helps ensure that industry standards, quality assurance benchmarks, and government regulations are met, Calibration is a standard process that involves: Y Checking the accuracy, precision, reliability, and deviation of the measurements produced by the instrument. Y Establishing the instrument's reliability Mapping any "drift" that occurs Y Setting the instrument to measure the needed range of values 2. Calibration of Glassware - Glassware is commonly calibrated using a liquid of known, specific density, and an analytical balance, The procedure is to determine the mass of liquid the glassware will hold, and to divide this mass of liquid by the density of the liquid, obtaining the corresponding volume of liquid 3. Calibration of following instruments : i. Analytical balance - Analytical balance calibration is best described as a process of checking the accuracy of a balance, often using a predefined calibration weight in the case of an anaytical balance with external calibration ii, pH meter - Clean the pH electrode with deionized water, and gently wipe it dry with Kimwipe, Place the pH electrode in the pH7 calibration solution. Press "yes" and wait until the reading shows that pH = 7 and ‘ready’, Press "yes" to confirm the calibration at pH7. aMODULE - 06 * Good Documentation Practice - 1, Writing of Laboratory Note Book - Weitting a laboratory notebook following Good Documentation Practice (GDP) is crucial for ensuiring accurate, reliable and traceable record of experintal work Here are some detailed guidelines and best practices for maintaining a laboratory notebook effectiovely > General P: Y Leability ciples of GDP in a laboratory Notebook - ¥ Permanence Y Timeliness ¥ Accuracy Y Tracebility > Structure of Laboratory Notebook entry - ¥ Tital and Date Y Objective or Purpose Y Materials and Methods ¥ Experimental Setup ¥ Observatiopn and Data Collection ¥ Result ¥ Discussion and Conclusion ¥ Signature and Witnessing > Additional Tips for Good Documentation - ¥ Use a page numbering system ¥ Avoid blank speces ¥ Use consistent language and Abbreviations Y Cross-Referencing ¥ Include error analysis 2. Preparation of Protocols - Creating detailed protocols for good documentation practices to ensure compliace with regulatory standards, maintain data integrity and facilitate consistent record- keeping, Here is a guidline on how to prepare these protocols: “> Good Documentation Practices (GDP) Protocols - Y Purpose ¥ Scope ¥ Responsibility Y General GDP Guidlines ¥ Document Creating Y Documentation type ¥ Record rferention and Storage ¥ Audit Trails and Data Integrity ¥ Training and Competency ¥ Review and Approval Process ¥ Handling Deviations and Discrepancies Y Periodic Audits and self-inspection . Preparation of Validation Reports - A validation report for good documentation practice provide document evidence that a documentation process or system performs as expected and complies with regulatory standards. Here’s comprehensive guid on how to preparev a validation report for GDP: > Validation Report for Good Documentation Practice (GDP) - Y Title page ¥ Purpose ¥ Scope ¥ Validation Teasm Y References ¥ Validation Approach Y Summary of Validation Activities ¥ Test Results and Observations ¥ Deviations and Corrective Action ¥ Summary and conclusion ¥ Recommendations Y Approval Signatures ¥ Appendices 45References . Title ~“Organic Spectroscopy” , Auther- “William Kemp” , edition 3rd, published by- “Bloomsbury publishing 2017” , ISBN 134915203X . Title “Instrumental Method of Chemical Analysis”, Author “Dr.B.K.Sharma”, publisher “Krishna prakashan media”, 1981, ISBN 8182830192 . . Title “Elementary Organic Spectroscopy” , Author- “Y R Sharma” , editionreprint , Publish by —“S. Chand publishing” 2007, ISBN 8181928842. Title — “Principle Of Instrumental Analysis , Authors — Douglas A. Skoog, James Holler, Stanley R. Crouch , Edition 7 , Publisher Cengage Learning , 2017. https://en. wikipedia org/wiki/High-performance_liquid_chromatography https://www.sciencedirect.com/science/article/pii/S 18785352 13001056 . https://ipe gov irvimages/GD-08-Good_Documentation_Practices.pdf