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Good Laboratory Practices for Pharmaceutical Quality Control Laboratories

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- Introduction - Requirements of GLP - Stepwise GLP in QC - Audit Findings - Issues due to not following GLP - Future Trends in GLP

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Good Laboratory Practices for Pharmaceutical Quality Control Laboratories

  1. 1. Good Laboratory Practices pharmauptoday@gmail.com
  2. 2. - Introduction - Requirements of GLP - Stepwise GLP in QC - Audit Findings - Issues due to not following GLP - Future Trends in GLP pharmauptoday@gmail.com Contents
  3. 3. Introduction
  4. 4. Introduction • Good Laboratory Practice (GLP) deals with the organization, process and conditions under which laboratory studies are planned, performed, monitored, recorded and reported. • GLP data are intended to promote the quality and validity of test data. • Weller gave an excellent practical explanation on what is expected from working in a regulated environment: “If experimental work is conducted in compliance with GLP, with or without the aid of computer, it should be possible for an inspector, maybe four or five years hence, to look at the records of the work and determine easily why, how and by whom the work was done, who was in control, what equipment was used, the results obtained, any problems that were encountered and how they were overcome”.
  5. 5. Introduction • Good laboratory practice requires testing normal and abnormal controls for each test atleast daily to monitor the analytical process. For the quality of a medicine sample to be correctly assessed: • The submission of a sample of an API, excipient or pharmaceutical product or a suspected counterfeit material to the laboratory, selected in accordance with national requirements, should be accompanied by a statement of the reason why the analysis has been requested. • The analysis should be correctly planned and meticulously executed. • The results should be competently evaluated to determine whether the sample complies with the specifications or other relevant criteria.
  6. 6. Introduction • Regulatory requirements, inspection and enforcement practices are quite dynamic. What is appropriate today may not need to be appropriate tomorrow. Regulations change but more often it is the inspection practices that change. • In the early 90’s the focus of inspections was on basic requirements of GLP and GMP, but then it changed to equipment hardware and later on to software and computer systems. • Today, the clear focus is on Quality control, Microbiology, Laboratory controls, data security, traceability and integrity of electronic records, driven mainly but not only by FDA’s regulation 21 CFR Part 11.
  7. 7. Requirements of GLP
  8. 8. Requirements of GLP
  9. 9. Requirements of GLP
  10. 10. Requirements of GLP
  11. 11. Stepwise GLP in QC
  12. 12. Stepwise GLP in QC. Lab Entry Lab Safety Lab Upkeep Reagents , Chemicals etc Instruments Lab Behavior Glassware Standards Procedures Computers Sampling Document ation Log Books Weighing Measurin g Volume Microbiology Stability Chromato graphy Sample Preparati on Investigations Lab Exit
  13. 13. Activity wise GLP in QC LAB MANAGEMENT Lab entry& exit Lab safety Lab upkeep Lab behavior RESOURCE MANAGEMENT People Procedures Reagents, Chemicals & Solvents Standards Glassware Instruments Log Books Computers QC ACTIVITIES Sampling Weighing Measuring volume Sample preparation Chromatography Microbiology Stability Documentation Investigations
  14. 14. Laboratory Entry
  15. 15. Laboratory Entry • There should be Standard Operating Procedure (SOP) for laboratory entry. • Laboratory entry should be restricted to authorised persons only. • Before entering the Lab ensure • Dress properly during a laboratory activity • Long hair, dangling jewelry, and loose or baggy clothing are a hazard in the laboratory. Long hair must be tied back, and dangling jewelry and baggy clothing must be secured. • Cut Nails, Shave your beard (for Gents) • Wash hands with soap solution • Wear proper gowning (Lab coat / Apron) • Wear Safety shoe & Shoes must completely cover the foot. • No sandals allowed in Laboratory. • Wear Goggles.
  16. 16. Laboratory Entry • The entry procedure may vary depending on the nature of the laboratory (e.g. Microbiology / QC laboratory of sterile plant / QC Laboratory of Solid dosages). • Laboratory should have complete laboratory layout at the entrance. • Do not allow any eatables / drinkables in to the laboratory. • Do not enter the laboratory in case of sickness. • Contact lenses may be not be worn in the laboratory.
  17. 17. Laboratory Health & Safety
  18. 18. Laboratory Health & Safety • All the persons who work in laboratory must undergo regular health checkups. • All the persons must be trained on health hazards associated with chemicals and protective measures. • Exposure to various chemicals / Drugs could cause a serious allergic reaction asthma, and other respiratory problems. • These MSDSs on Infectious Agents are organized to contain health hazard information such as infectious dose, viability (including decontamination), medical information, laboratory hazard, recommended precautions, handling information and spill procedures.
  19. 19. Laboratory Health & Safety • Before entering the laboratory it is mandatory to know the lab safety procedures thoroughly. • All individuals must be trained on handling PPEs (Personal Protective Equipments) and Lab Safety. • All must know the availability & usage of • Emergency Exit & Assembling point • Fire extinguishers & Fire Alarm • Emergency showers • Eye washers • First Aid kit etc. • Report any accident (spill, breakage, etc.) or injury (cut, burn, etc.) to the supervisor immediately, no matter how trivial it seems.
  20. 20. Laboratory Health & Safety • If you or your lab partner is hurt, immediately (and loudly) yell out the others attention. Do not panic. • If a chemical splash in your eye(s) or on your skin, immediately flush with running water for at least 20 minutes. • Avoid using electric extension cords whenever possible. If you must use one, obtain a heavy- duty one that is electrically grounded, with its own fuse, and install it safely. Extension cords should not go under doors, across aisles, be hung from the ceiling, or plugged into other extension cords. • When using compressed air, use only approved nozzles and never direct the air towards any person. • Exercise care when working with or near hydraulically- or pneumatically-driven equipment. Sudden or unexpected motion can inflict serious injury.
  21. 21. Laboratory Health & Safety • Treat every chemical as if it is hazardous. • Make sure all chemicals are clearly and correctly labeled with the substance name, concentration, date, and validity. • Never return chemicals to reagent bottles. • Never allow a solvent to come in contact with your skin. Always use gloves. • Never pipette by mouth; use a bulb. • Clean up spills immediately. • Heated glassware remain very hot for a long time. They should be set aside in a designated place to cool, and picked up with caution. Use tongs or heat protective gloves if necessary.
  22. 22. Laboratory Health & Safety • Do not work alone in the laboratory. • Do not touch the reagents / solvents / standards / drugs with bare hands. • Do not smell any reagent / solvent / standard / drug. • Do not run / rush in the laboratory. • Do not keep any article which blocks the passage / emergency exit. • Do not lift the weights / heavy bottles suddenly • Do not leave an on-going experiment unattended. • Do not handle broken glass with your bare hands. Use a brush and dustpan to clean up broken glass. Place broken glass in the designated glass disposal container.
  23. 23. Laboratory Health & Safety • Stop your work, or that of your colleagues — when you are concerned or suspicious of danger or unnecessary risk. • Keep the poisonous chemicals and reagents under lock & key and usage record shall be maintained. As per USP General Notices & Requirements:
  24. 24. Laboratory Upkeep
  25. 25. Laboratory Upkeep • Keep your lab space clean and organized. • Maintain unobstructed access to all exits, fire extinguishers, electrical panels, emergency showers, and eye washes. • Do not use corridors for storage or work areas. • Do not store heavy items above table height. Any overhead storage of supplies on top of cabinets should be limited to lightweight items only. Also, remember that a 36" diameter area around all fire sprinkler heads must be kept clear at all times. • Areas containing lasers, biohazards, radioisotopes, and carcinogens should be posted accordingly. However, do not post areas unnecessarily and be sure that the labels are removed when the hazards are no longer present. • Clean your lab bench and equipment.
  26. 26. Laboratory Upkeep • Know the locations and operating procedures of all safety equipment. • Clean up spills immediately. • Use volatile and flammable compounds only in a fume hood. Procedures that produce aerosols should be performed in a hood to prevent inhalation of hazardous material. • Make sure all chemicals are clearly and currently labeled with the substance name, concentration, date, and name of the individual responsible. • Guards on machinery must be in place during operation. • Make sure that acids and bases are not stored together. If there were to be a spill, chemical reactions could occur if they are stored in the same storage area.
  27. 27. Laboratory Upkeep • Make sure you check the MSDS of the acid or base to determine you are not storing incompatible chemicals together. • Be aware that all containers MUST be properly labeled so that identification can be achieved as easy as possible. • Pay close attention to expiration dates, making sure that the acid or base is disposed of on or before the expiration date. • Acids and bases should be stored in a Corrosive Cabinet. • Obtain and read the most current Materials Safety Data Sheet (MSDS) for the specific acids and other chemicals that you will be using.
  28. 28. Laboratory Upkeep • Chemicals should not be stored in the lab haphazardly. Many chemicals are incompatible with others and improper storage can be quite dangerous. • Organic compounds and inorganic compounds should be stored separately as well. • Many compounds have specific storage requirements and these are often listed on the label. • The most common source of mercury in a lab is a broken thermometer. Once a thermometer has been broken, quick and thorough clean up is required. Mercury is quite volatile and the health effects are not trivial. • Corrosive liquids (e.g. mineral acids, alkali solutions and some oxidizers) represent a very significant hazard because skin or eye contact can readily occur from splashes and their effect on human tissue generally takes place very rapidly. Bromine, sodium hydroxide, sulfuric acid and hydrogen peroxide are examples of highly corrosive liquids.
  29. 29. Laboratory Upkeep • Liquid corrosives should be stored below eye level. • Corrosive gases and vapors are hazardous to all parts of the body; certain organs (e.g. the eyes and the respiratory tract) are particularly sensitive. The magnitude of the effect is related to the solubility of the material in the body fluids. Highly soluble gases (e.g. ammonia, hydrogen chloride) cause severe nose and throat irritation, while substances of lower solubility (e.g. nitrogen dioxide, phosgene, sulfur dioxide) can penetrate deep into the lungs. • Regulators and valves should be closed when the cylinder is not in use and flushed with dry air or nitrogen after use. • When corrosive gases are to be discharged into a liquid, a trap, check valve, or vacuum break device should be employed to prevent dangerous reverse flow.
  30. 30. Laboratory Upkeep • Corrosive solids, such as sodium hydroxide and phenol, can cause burns to the skin and eyes. Dust from corrosive solids can be inhaled and cause irritation or burns to the respiratory tract. Many corrosive solids, such as potassium hydroxide and sodium hydroxide, can produce considerable heat when dissolved in water. • Wear gloves and eye protection when handling corrosive solids. • When mixing with water, always slowly add the corrosive solid to water, stirring continuously. Cooling may be necessary. • If there is a possibility of generating a significant amount of dust, conduct work in a fume hood. • Use bottle top dispensers wherever required.
  31. 31. Laboratory Upkeep • Adequate lighting is necessary in the Laboratory to perform the activities. • Light sensitive drugs can be affected by sunlight (ultraviolet light) or by artificial light (like fluorescent light). Proper precautions shall be taken to store, handle the Light sensitive products. • The Room Temperature & the Humidity shall be controlled and monitored in the laboratory. • The area of the sample storage in the laboratory shall be mapped for temperature and documented appropriately. • The samples which are supposed to store at other temperature (Refrigerated / Freezer) shall be stored appropriately.
  32. 32. Laboratory Upkeep - 5 S principle • Follow 5 S principle in the laboratory. • The 5S system is a tool that supports an operating philosophy that emphasizes discipline, efficiency, and attention to detail 1.Sort 2.Set in Order 3.Shine 4.Standardize 5.Sustain • Benefits of 5S • Safety • Employee involvement • Organization/Efficiency • Cleanliness • Space Savings • Lower inventories • More consistent experiments • Faster on-boarding of new employees • Timely delivery of supplies • Additional funding
  33. 33. Laboratory Upkeep - 5 S principle 1. Sort • Document baseline work area ―before‖ condition (pictures, audits, etc.) • Perform the initial cleaning of the work area • Separate the necessary items from unnecessary items • Remove unnecessary items from the work area
  34. 34. Laboratory Upkeep - 5 S principle 1. Sort • Sorting Methodology: • Establish a holding area for items (Red tag area) • Identify items not required at the • current location, if at all • Team members evaluate items in Red Tag area • Dispose of items with team approval
  35. 35. Laboratory Upkeep - 5 S principle 2. Set in Order • Identify necessary tools, equipment, supplies, etc. • Determine location for necessary items • Designate & outline permanent locations for items
  36. 36. Laboratory Upkeep - 5 S principle 2. Set in Order How to set in order • Define the major processes performed • For each process, draw a map of the locations of each activity • Identify sources of waste and corrective measures • Position items where they are needed based on frequency
  37. 37. Laboratory Upkeep - 5 S principle 3. Shine • Clean area thoroughly – Make it shine! • Clean & inspect areas daily
  38. 38. Laboratory Upkeep - 5 S principle 3. Shine • Machines & Equipment • Fix leaks & other problems • Prevent contamination • Inspect daily • Fix potential issues before they become major problems
  39. 39. Laboratory Upkeep - 5 S principle 4. Standardise • Develop SOPs • Document equipment & processes • Develop audit schedule and assign tasks to individuals • Establish a methodology to ensure consistent strategy for 5S implementation
  40. 40. Laboratory Upkeep - 5 S principle 5. Sustain • Establish procedures on 5 S. • Maintain established procedures. • Ensure 5 S is a long term, laboratory-wide goal. • Analyze results, hold team meetings and train scientists on the importance of 5 S.
  41. 41. Laboratory Behavior
  42. 42. Laboratory Behavior • Scientists shall behave professionally in the laboratory. • Observe keenly each step of the experiment. • If errors are obvious, such as the spilling of a sample solution or the incomplete transfer of a sample composite, the analyst should immediately document what happened. • Scientists should not knowingly continue an analysis they expect to invalidate at a later time for an assignable cause (i.e., analyses should not be completed for the sole purpose of seeing what results can be obtained when obvious errors are known). • Analysts should check the data for compliance with test specifications before discarding test preparations or standard preparations. • Do not shake hands while working. • Do not use mobile phones in the laboratory.
  43. 43. People
  44. 44. People • People should undergo periodic health check-ups. • Be able to confirm that employees are qualified for their positions by having a current CV and Job description on file. • All employees should update periodically (annually, biannually, etc.) and as responsibilities change (for example, after a promotion). • Have accurate training records on file that are easily retrieved • Conduct frequent refresher training on quality and safety topics
  45. 45. Instruments
  46. 46. Instruments • All the critical instruments (new & modified) must be Validated (IQ/OQ/PQ) and Calibrated. • Written procedures must be established to operate / Calibrate the instruments. 21 CFR 211.160 (b) (4) :
  47. 47. Instruments • All instruments must be numbered appropriately for easy identification. • All instruments must be labeled with Calibration status & due date. • Maintain schedules for Preventive maintenance & Calibration activities of instruments. • All the instruments must have usage and maintenance log books. • Schedules and procedures (including assignment of responsibility) should be established for the preventative maintenance of equipment.
  48. 48. Instruments • Equipment calibrations should be performed using standards traceable to certified standards, if existing. • Records of these calibrations should be maintained. • Instruments that do not meet calibration criteria should not be used. • Deviations from approved standards of calibration on critical instruments should be investigated to determine if these could have had an impact on the quality analysed using this equipment since the last successful calibration.
  49. 49. Reagents, Chemicals & Solvents
  50. 50. Reagents, Chemicals & Solvents • All reagents and solutions in the laboratory areas shall be labeled to indicate identity, titer or concentration, storage requirements, and expiration date. • Deteriorated or outdated reagents and solutions should not be used. • If reagents and solutions used for non-regulated work are stored in the same room as reagents for regulated studies, all reagents must be labeled. • Reagents that are not adequately labeled, even if not intended for use in regulated studies, may have an adverse effect on regulated laboratory work. It is also good practice to include the Date opened. This can be critical for some chemicals such as ether.
  51. 51. Reagents, Chemicals & Solvents • The expiration date depends on the nature of the chemical. E.g. Sodium chloride has practically no expiration date. In these cases it might be acceptable to indicate NONE or Not applicable (N/A) on the label for expiration date. • The laboratory must be prepared to justify this designation. Formal studies are not required to justify assigned expiration dates. • It is sufficient to assign expiration dates based on literature references and/or laboratory experience. • The label should indicate special environmental conditions, for example Refrigerate or Protect from light. • Do not mix sulphuric acid with water inside a cylinder. • Check the conductivity / pH of the water before use.
  52. 52. Glassware
  53. 53. Glassware • Use Class A Glassware only for measurement. • Validate the glassware / vial cleaning procedure. • Do not use broken glassware. • Verify & Preserve the Certificate provided by the vendor for glassware. • Do not heat the glassware that is cracked, chipped or scratched.
  54. 54. Standards
  55. 55. Standards • Certified reference standards can be purchased from appropriate suppliers. • If standards are not available, the recommendation is to take a lot of your own material, and analyze, certify and use it as the standard. • Sometimes certified standards are too expensive for day-to-day routine use. In this case in-house laboratory standards can be used as working standards. However, they should be made from high purity material and be compared against the primary standard to ensure the traceability chain. For the comparison, validated test methods should be used. • All reference material, either purchased or in-house, should be subject to a quality control procedure. • This includes regular checks of purity, identity and concentrations.
  56. 56. Standards • Standards should be stored in their original stoppered container away from heat and humidity and protected from light • Store up to the validity of current lot. • USP RS should be stored as per the label directions. • Do not dry reference standards in their original container. Dry only the required quantity of the standard on a glass Petridish, whenever specified. • Do not put back any excess quantity of the dispensed standard back into the stock bottles. • Allow the container of the standard to attain room temperature before using. Wipe the outer surface of the container with tissue paper to remove moisture. As per USP General Notices & Requirements:
  57. 57. Procedures
  58. 58. Procedures • These procedures describe the methods that will be used to implement and perform the stated policies. • The procedures define who should perform the specific tasks, when the task should be done, and where the documentation will be made showing that task was performed. • The Procedure (Standard Operating Procedures) shall be reviewed periodically. • SOPs should preferably be written in the laboratory close to the instrument, and not in an office. It should be either written or thoroughly reviewed by the instruments’ operators. • SOPs should not be written to explain how procedures are supposed to work, but how they work.
  59. 59. Procedures • This ensures that the information is adequate and that the document invites rather than discourages routine use. • Content should cover: • SOP unique number and revision number, • Page number and total number of pages, • for equipment testing: performance acceptance criteria, recommended corrective actions, and a template for continuous entries of test results and corrective actions, • printing history.
  60. 60. Procedures • Copies of SOPs for equipment should be located close to the instruments and must be easily accessible by operators. • Deviations from SOPs in a study must be authorized and significant changes in established SOPs must be authorized in writing by management. • Standard operating procedures should be drafted in a language understood in the workplace.
  61. 61. Procedures • Have approved procedures in place for • Method validation and transfer requirements. • Instrument qualification, calibration and preventive maintenance. • Instructions for qualification and calibration include the actions to be taken when an instrument fails calibration. • Out of specification/out of trend results, deviations and investigations include some type of root cause analysis steps to help identify the cause of the issues and prevent it from occurring in the future. • The use and security of lab software, including data acquisition software and LIMS (Laboratory Information Management System). • To define how raw data and lab documentation is handled. • To define the process for receiving samples and how samples are managed within the lab.
  62. 62. Log Books
  63. 63. Log Books • Laboratory equipment log books are important records of machine calibration, verification checks, cleaning, maintenance and use. • Always record entries legibly, neatly and in permanent ink. • All log books require one signature per line entry. • All entries should note the date and time of the entry / observation. • Never leave blank spaces and never erase or remove material added. Draw lines through any blank spaces at the same time of making entries. • Never remove pages from the log book. • Entries in the log book shall be in chronological order.
  64. 64. Log Books • Logbooks should be kept near the instrument so it is easy to make entries or to check. • Laboratory equipment log books should show the following • Date • Time • Product Name • Batch Number • Initials (Sign) of the performer
  65. 65. Documentation
  66. 66. Documentation Policy The first tier of documentation is the policy manual. This is the document that defines what will be done and why. A quality policy manual should be written so it is clear, precise and practical, and easy to understand. The why can be stated just once as a quality policy statement. This statement should be a short, simple definition of the organization’s quality intentions.
  67. 67. Documentation Procedures • The second tier of documentation is quality procedures. These procedures describe the methods that will be used to implement and perform the stated policies. The procedures define who should perform the specific tasks, when the task should be done, and where the documentation will be made showing that task was performed. Work Instructions • Work instructions are usually department, machine, task, or product oriented an spell how a job will be done. The instructions are the most detailed of the documentation hierarchy. A work instruction may be in the form of a detailed drawing, recipe, routing sheet, specific job function or simply a sample for comparison or conformity. Records • Records are a way of documenting that the policies, procedures, and work instructions have been followed. Records may be forms that are filled out, a stamp of approval on a product, or a signature and date on some type of document, such as routing sheet. Records are used to provide traceability of actions taken on a specific product or batch of products. They provide data for corrective actions and a way of recalling products, if necessary.
  68. 68. Documentation Raw Data • Raw data refers to any laboratory worksheets, records, memoranda, notes, or exact copies thereof, that are the results of original observations and activities of a study. The term covers all data necessary for the reconstruction of the report of the study. Raw data may include handwritten notes, photographs, microfiche copies, computer print-outs, magnetic media, dictated observations, and electronically recorded data from automated instruments. Examples include results of environmental monitoring, instrument calibration records, and integrator output from analytical equipment. Raw data may also be entries in a worksheet used to read and note information from the LED display of an analytical instrument.. Laboratory Note book • For raw data entries, it is recommended to use controlled forms or a laboratory notebook for each study. This should be robust, bound and have the pages numbered. All entries should be made in indelible ink. Scientists and technicians sometimes record raw data on scraps of paper or even on paper towels. Their intention is to neatly transcribe the information to official data forms at a later time and to discard the originally recorded data. This practice should be discouraged, because the scraps of paper are the real raw data, and must be retained. More recently electronic notebooks are used instead of paper notebooks. For US-FDA GLP/GMP regulated laboratories the regulation on electronic records and signatures, 21 CFR Part11 applies.
  69. 69. Documentation Specifications & Analytical Methods • Use only validated analytical methods for testing. • Use only approved Specifications and testing procedures. Standard Operating Procedures • There should be standard operating procedure for all critical activities / operation and calibration of instruments. • Follow Standard operating procedures.
  70. 70. Computers
  71. 71. Computers • Identify each user uniquely • Implement adequate password controls • Establish different user roles / access privileges • Establish and maintain a list of current and historical users • Control changes to the system • Use only trained staff to operate the system • Understand predicate rules for laboratory records • Define and document e-records for the system • Review the audit trails for each batch set • Back the system up regularly
  72. 72. Sampling Spatulas for solids Dip tubes for liquids
  73. 73. Sampling • Sampling tools should be available to the sampler, e.g. to open containers (knives, hammers,...), material to reclose the packages (sealing tape), self-adhesive labels to indicate that some of the contents have been removed, etc... • Sampling tools should be made of inert materials (e.g. polypropylene or stainless steel; avoid glass) and kept very clean. After use, thoroughly washed, rinsed with water or suitable solvent, dried and stored in clean conditions. • Disposable sampling materials can also be used. • Washing facilities should be located in, or close to, the sampling area. • Cleaning procedure should be documented and validated (= demonstrated efficiency). • Sterile pharmaceutical products should be sampled under aseptic conditions.
  74. 74. Sampling • Sampling operations should be supervised and documented => sample collection form => always kept together with the collected sample. • Sample collection form: written record of the sampling operations, containing: batch number, sampling date/place, reference to sampling protocol used, description of containers and materials sampled, possible abnormalities, any relevant observations, name/signature of the sampler... • Store the sample in a properly labelled container: sample type, name of material, identification code, batch number, code, quantity, date of sampling, storage conditions, handling precautions, container number. Rooms for sample storage: • Security and adequate storage conditions (light, ventilation, safety requirements, and any special requirements) should be ensured for the rooms in which samples are stored. • Samples should be stored according to the storage conditions as specified for the respective API, excipient or drug product. • Packaging materials similar to those in which the bulk is supplied should be used for long-term storage.
  75. 75. Weighing
  76. 76. Weighing • Selection of the suitable balance based on the definition as well as considering other balance characteristics such as balance readability, minimum weight, capacity, accuracy, built-in calibration and uncertainty values. • Use Qualified & Calibrated analytical balance. • Weighing result, as obtained during an analysis period is influenced by a series of factors relating to balance and ambient conditions. However, one should also consider sample features during weighing process. Samples that are liquids, can undergo process of evaporation. • A factor reverse to evaporation is absorption of moisture from ambient air by a sample. it is very important in case of hygroscopic samples. • The effect of moisture absorption is differences in mass determination, each measurement will have higher mass readout than the previous one. For the purpose of proper weighing of such substances, weighing vessel should be clean and dry.
  77. 77. Measuring Volume
  78. 78. Measuring Volume • Volume may be measured reliably with a pipette, a burette, or a volumetric flask. • Volumetric equipment is marked by the manufacturer to indicate not only the manner of calibration (usually TD for "to deliver" or TC for "to contain") but also the temperature at which the calibration strictly applies. Pipettes and burettes are ordinarily calibrated to deliver specified volumes, whereas volumetric flasks are calibrated on a to-contain basis. • TC pipettes deliver all the volume including the tip, and then must be "blown out" to get the last drop. Blowing out is done using the associated pipette pump, NEVER your mouth. • A meniscus is the curved surface of a liquid at its interface with the atmosphere.
  79. 79. Measuring Volume Reading a burette / pipette: • The tips of two styles of measuring pipettes. • The Mohr pipette is shown on the left, the graduation lines stop short of the tip, • The serological pipette on the right, the graduation lines, pass through the tip. X √ X
  80. 80. Sample Preparation
  81. 81. Sample Preparation Sonication: • Frequently change the solution in the bath. Solutions that become contaminated with particles that settle on the bottom of the bath will decrease ultrasonic activity. • Do not place sample containers directly on the bottom of the bath. Containers on the bottom of the bath will decrease cavitation and will damage the transducer because they reflect the ultrasonic energy back to the transducer. • Containers should be placed in an open mesh basket or in an insert tray or be suspended in the solution to position the container in the optimal zone of the bath and to allow the ultrasonic waves to penetrate around the containers. • Degas the solution in the bath by sonicating for 5–10 min to enhance cavitation prior to sonicating samples. • Sonication often results in heating of sample solutions. If using volumetric flasks, sonicated sample solutions should be allowed to cool to room temperature before they are diluted to volume. Since sonication induces localized areas of high temperature, this may adversely affect thermally labile compounds.
  82. 82. Sample Preparation Eliminating Insoluble Particles: • Centrifugation shall be performed as specified in the validated method for specified time. • Filter membrane selection based on analyte and solvent properties, so the filters shall be used as specified in the validated method. • Filters with pore size distribution around 0.45 μm are suitable for most applications, although high efficiency LC columns with particles of < 3 μm usually require filtration of particles of ≤ 0.2 μm. • It is important that the selected filter is validated during the validation exercise to ensure that the active or related impurities are not lost to the filter and that no significant interferences are generated by the selected filter. • As part of this process, one needs to study the effect of wetting the filter prior to sample collection. Usually the first 5 mL of the eluate are discarded to obtain acceptable and reproducible recoveries.
  83. 83. Chromatography
  84. 84. Chromatography • All reagents and solvents should be of the highest quality. HPLC grade reagents may cost slightly more than lower grade reagents, but the difference in purity is marked. • HPLC grade reagents contain no impurities to produce spurious peaks in a chromatogram baseline. • Always use Purified water / Milli Q water as both inorganic and organic contaminants are removed. • Do not use RO water / de-ionsed water for HPLC. • Due to existence of impurity, deionized water shows higher absorption
  85. 85. Chromatography Buffers used for HPLC • All the solutions should be clear, homogenous & free from particulate matter. • Buffer solutions must be filtered through 0.45μm filter or as specified in the method by using Solvent Filtration Kit. • Do not keep mobile phase under the suction for more than 4 to 5 minutes as the composition of mobile phase change especially if highly volatile solvents are used. • Buffer solutions must not be left in system to avoid crystalisation. • effect on pump -- damage plunger and seal • effect on column -- creation of column voids • effect on flow line -- corrosion of stainless steel lines • Possible bacterial growth especially phosphate buffers -- good medium for bacterial and fungus growth. • Ideally, solutions should be prepared fresh everyday
  86. 86. Chromatography Mobile Phase for HPLC • Always cover the container • To prevent evaporation of solvents • To prevent dust enter the mobile phase • To reduce the vapors in the room • Do not expose the bottle to direct sunlight/wind. • Always use a suction filter to Prevent particulates from reaching pumps Stationary phase (Column) for HPLC • Avoid pressure shocks on the column. • Pressure shocks lead to channeling in the column, which results in peak splitting in corresponding chromatogram. • Always keep both ends of the column closed, after usage. • Keep the columns in the designated column cabinets after use.
  87. 87. Chromatography • The solvent lines should be clean, growth-free and should have no sharp bends or creases in them. • Solvent reservoirs should be placed as high as possible on or in the instrument – always higher than the pump inlet manifold. • Before attempting any solvent change, ensure that the solvent already in the system and column is compatible with the new solvent. • The column efficiency, capacity factor etc…should be measured at the start and end of the clean-up procedure to ensure that it has been performed successfully and has improved the column performance. • Ensure that no buffers/samples are present on the column and that the solvent used prior to the clean-up is miscible with the first wash solvent. • After the clean-up, ensure that the test mobile phase is compatible with the last solvent in the column. • Samples require filtration if they contain suspended solids. This can be performed on-line using a pre-column filter or as the sample is introduced to the vial.
  88. 88. Microbiology
  89. 89. Microbiology • Good laboratory practices in a microbiology laboratory consist of activities that depend on several principles: aseptic technique, control of media, control of test strains, control of equipment, diligent recording and evaluation of data, and training of the laboratory stuff. • Clean spills immediately according to established protocols and disinfect the area thoroughly • Keep bench top uncluttered • Minimize traffic and unnecessary movements around the work area (movement can stir up air currents which can carry contaminants into the work area). • All work with infectious material should be carried out in a specific area, and the material should not be carried throughout or out of the lab unless in a closed or capped container. • Minimize aerosol generation; if unavoidable, activities should be carried out in a biological safety cabinet
  90. 90. Microbiology • Use proper aseptic technique for the transfer and handling or microorganisms and instruments • Keep sterile and non-sterile objects separate • Minimize exposure to outside air (i.e. keep lids off sterile containers for as little time as possible) • Avoid contact with non-sterile surfaces and items (i.e. never place lids/caps onto a work surface, hold lids in an opening-down position), and • Hold open containers at an angle whenever possible to prevent contaminants from falling in. Media, Reagents Preparation, Storage, Qualification: • It’s important to choose the correct media or components in making media based on the use of accepted sources or references for formulas. • The LAL reagentis critical in Endotoxin testing, and must be well controlled. • When the LAL reagent (lot) changes, the Micro Lab must confirm the labeled LAL reagent sensitivity λ (EU/ml). This is to re-calculate the Maximum Valid Dilution (MVD), which is the maximum allowable dilution of a sample to determine the endotoxin limit.
  91. 91. Microbiology Reference Standards: • Microbial cultures are delicate standards. Procedures should specify careful handling instructions. • Preparation and resuscitation of cultures should follow the instructions of the supplier or a validated, established method. • USP recommends using the "Seed-Lot" technique for storage of stock cultures, i.e., using working cultures and never returning unused passages back to original stock. • In addition, there should be an established maximum number of passages (5 or less), and maximum storage time for working cultures. • Cultures for use in compendial tests should be acquired from a national culture collection, in frozen, freeze-dried, on slants, or in ready-to-use forms. • Confirmation of the purity and the identity should be performed prior to its use in quality control testing. • Ready-to-use cultures may require additional confirmation of inoculums size.
  92. 92. Microbiology Laboratory Operations/Housekeeping: • Safety first. The Micro Lab should practice aseptic techniques during testing in general, to avoid microbial contamination and false positives. • In the Micro Lab, areas where EM, water, or product samples are handled/incubated must be adequately separated from areas where there are tests that involve live cultures or sub-culturing, microbial ID or investigations. • Housekeeping must be properly maintained to prevent use of expired or contaminated testing mat’s. • Verify cleanliness of work stations, cleared of extraneous or previous test residue, prompt removal of refuse, and clean utensils and equipment
  93. 93. Stability
  94. 94. Stability
  95. 95. Stability • Establish approved procedures for • Conducting Stability studies • Withdrawal of Stability studies • Investigation of deviations, incidences, OOT and OOS results. • Lay down a properly designed test protocol. • Conduct the stability study as per the protocol. • Make estimate of shelf life from the generated data. Display and Recording of Set Values • Digital display and recording through chart recorders, data loggers, computer and software • Alternatively or in addition, memory back-up with provision of downloading of data at any time
  96. 96. Stability • Delays are needed to be provided in alarms – to avoid unnecessary notification, door opening condition is not registered unless the door is open for more than five minutes. In case of fluctuation, the system allots say around 20 min for system to re-equilibrate before setting an alarm. • The alarm might be cleared only when a user ID and password is entered. • Prepare samples and keep at the selected storage conditions • Withdraw at the prescribed sampling intervals and subject to analysis. • Complete the testing of stability samples within specified time period. • Monitor the temperature & Humidity of the stability chambers. All the fluctuations shall be corrected, documented and justified.
  97. 97. Investigations
  98. 98. Investigations
  99. 99. Investigations • Investigations of "Out of Specification (OOS) / Out of Trend (OOT)/ Atypical -results" have to be done in cases of: – Batch release testing and testing of starting materials. – In-Process Control testing: if data is used for batch calculations/decisions and if in a dossier and on Certificates of Analysis. – Stability studies on marketed batches of finished products and or active pharmaceutical ingredients, on-going / follow up stability (no stress tests) – Previous released batch used as reference sample in an OOS investigation showing OOS or suspect results. – Batches for clinical trials. • All solutions and reagents must be retained until all data has been second person verified as being within the defined acceptance criteria.
  100. 100. Investigations • Pharmacopoeia have specific criteria for additional analyses of specific tests (i.e. dissolution level specification for S1, S2 & S3 testing; Uniformity of dosage units specification for testing of 20 additional units; Sterility Testing). • However if the sample test criteria is usually the first level of testing and a sample has to be tested to the next level this should be investigated as it is not following the normal trend. • Unexpected results and/or events should be documented and thoroughly investigated. • It is not sufficient to determine that additional training is required for the analyst or glassware wasn’t adequately cleaned. Regardless of the method used, it the investigation steps and results need to be clearly documented. Include supporting documentation so that anyone reading the investigation report understands the actions taken and the corresponding results. In the event that the cause cannot be identified, be sure the investigation report illustrates that adequate measures were taken to attempt to identify the cause. • Don’t stop at identifying the root cause. Put a plan in place to resolve the issue in the current situation and prevent it from occurring in the future, and then follow through to initiate the plan and evaluate if it has the desired results.
  101. 101. Laboratory Exit
  102. 102. Laboratory Exit • There should be a defined procedure for lab exit. • There should be evacuation plan defined in safety manual. • Before coming out of the Lab ensure • Switch off unnecessary instruments. • Remove / Dispose the apron, Cap and gloves. • Wash hands with soap solution.
  103. 103. Audit Findings
  104. 104. Canada Audit citations • Insufficient testing of raw material. • Incomplete specifications. • Specifications not approved by QC. • Test methods not validated. • Use of raw material after retest date without proper retesting. • Use of raw material after the expiration date. • Incomplete validation of test methods. • No SOP for conditions of transportation and storage.
  105. 105. Canada Audit citations • Evidence of falsification or misrepresentation of testing results / forgery of COA. • Non-compliant products made available for sale. • Incomplete/inadequate specifications. • Finished product specifications not approved by QC. • Incomplete testing. • Lack of or insufficient validation of test methods. • Inadequate method transfer for a validated analytical method. • Method validation report does not specify the revision of the analytical method used at the time of validation.
  106. 106. US FDA Audit Findings – Laboratory Controls • Failure to maintain laboratory control records with complete data derived from all tests conducted to ensure compliance with established specifications and standards, including examinations and assays. • Your firm failed to establish laboratory controls that include scientifically sound and appropriate specifications, standards, sampling plans, and test procedures designed to assure that components, drug product containers, closures, in-process materials, labeling, and drug products conform to appropriate standards of identity, strength, quality, and purity (21 CFR 211.160(b)). • Your firm failed to ensure that laboratory records included complete data derived from all tests necessary to assure compliance with established specifications and standards (21 CFR 211.194(a)). • Your laboratory’s written procedure failed to establish proper retesting practices for out-of-specification results.
  107. 107. US FDA Audit Findings – Laboratory Controls • Your firm failed to thoroughly investigate any unexplained discrepancy or failure of a batch or any of its components to meet any of its specifications, whether or not the batch has already been distributed (21 CFR 211.192). • Your firm failed to investigate and document out-of-specification results according to a procedure. • Your laboratory’s written procedure failed to establish proper retesting practices for out-of-specification results. • Recognizing that an Out-of-Specification Test Result was obtained in [REDACTION] specification of [REDACTION] as per requirement of your Quality Control/Quality Assurance Standard Operating Procedure for OOS Investigation, SOP Number 2802, Reporting, Investigation and Disposition of Out of Specification (OOS) Laboratory Results, effective 02/05/09 and previous revisions, it appears that you avoided to perform an Investigation for the assay test result of [REDACTION]. • The Quality Unit is not initiating investigations and documentation of those investigations in a timely manner.
  108. 108. US FDA Audit Findings – Laboratory Controls • Your firm does not have adequate written procedures for production and process controls designed to assure that the drug products you manufacture have the identity, strength, quality, and purity they purport or are represented to possess [21 C.F.R. § 211.100(a)]. • Failure to have an API stability program to monitor stability characteristics of your firm’s APIs, and failure to set an expiry or retest date for APIs based on the evaluation of data derived from stability studies. • Your laboratory’s written procedure failed to establish proper retesting practices for out-of-specification results. • Your firm failed to establish and follow an adequate written testing program designed to assess the stability characteristics of drug products and to use results of such stability testing to determine appropriate storage conditions and expiration dates (21 CFR 211.166(a)). • Your firm’s quality control unit failed to review and approve all drug product production and control records to determine compliance with all established, approved written procedures before a batch is released or distributed (21 CFR 211.192).
  109. 109. US FDA Audit Findings – Microbiology • Your firm failed to establish and follow appropriate written procedures that are designed to prevent microbiological contamination of drug products purporting to be sterile, and that include validation of all aseptic and sterilization processes (21 CFR 211.113(b)). • Your firm failed to establish an adequate system for monitoring environmental conditions in aseptic processing (21 CFR 211.42(c)(10)(iv)). • Your firm failed to establish maximum holding times for vials used in media fills, prior to incubation
  110. 110. US FDA Audit Findings – Microbiology • Your firm failed to establish and follow appropriate written procedures that are designed to prevent microbiological contamination of drug products purporting to be sterile, and that include validation of all aseptic and sterilization processes (21 CFR 211.113(b)). • Your firm failed to establish an adequate system for monitoring environmental conditions in aseptic processing (21 CFR 211.42(c)(10)(iv)). • Your firm failed to establish maximum holding times for vials used in media fills, prior to incubation
  111. 111. Issues due to not following GLP
  112. 112. Issues due to not following GLP • Accidents / Incidents • Health Hazards Lack of Lab Safety • Time delay for searching • Contamination & no clarity of work Lack of Lab Upkeep • Accidents / Incidents • Erratic results Lack of Lab Behavior • Time delay for repetitions & instrument failures • Erratic results Lack of Instrument maintenance • Accidents / Incidents • Erratic results & Contamination Lack of Reagent maintenance
  113. 113. Issues due to not following GLP • Erratic volume measurement & wrong results • Contamination Lack of Glassware maintenance • Wrong / Erratic results • Contamination Lack of Standard maintenance • Non Uniformity of performing activities & results • Perform activities wrongly Lack of Procedures • No control on usage of instrument • No evidence of usage of instrument Lack of Log Books • Evidence of performing the activity • To control errors / mistakes Lack of Documentation
  114. 114. Issues due to not following GLP • Loss on data (security) • Misuse / deletion of data Lack of Computer maintenance • Spillage can lead to wrong / Erratic results • Contamination & mix-up Not following Sampling instructions • Erratic / wrong results • Time delay to perform investigations Not weighing properly • Erratic volume measurement & wrong results • Time delay to perform investigations Lack of correct volume measurement • Erratic / Wrong results • Time delay to perform investigations Not following sample preparation technique
  115. 115. Issues due to not following GLP • Instrument error / malfunction / breakdown • Erratic results • Cost effective • Time delay for investigations Lack of Good Chromatography • Wrong / Erratic results • Contamination & mix-up Lack of Good Microbiological practices • Erratic / wrong time study • Wrong establishment of Retest / Expiry date Not following proper stability practices • Erratic volume measurement & wrong results • Repetitive issues due to no or incorrect root cause. Inadequate Investigations
  116. 116. Issues due to not following GLP Summary of Issues due to not following GLP in QC laboratories • Accidents / incidents & Health hazards • Wrong / errors in volume or weight measurement leads to erratic / wrong results • Some errors may be identified some may not be identified • Instrument errors / malfunctions • Contamination of Sample, Reagent, Solvent / product • Waste of time due to investigation of unexpected / unknown cause. • No evidence / documental proof of data generated. • Increase in cost due to repetitions & investigations • Due to wrong result generation impacts the quality / purity of the product • All the errors generated knowingly / unknowingly can effect patient safety. • Regulatory agency action results in 483’s, Warning letters, Import alerts, Non-Compliance reports etc.
  117. 117. Future Trends in GLP
  118. 118. Future Trends in GLP • The Good Laboratory Practices shall be followed strictly in the Quality control laboratories of all analytical laboratories i.e. in pharmaceutical QC lab, Analytical R &D as well as contract testing laboratories. • GLP also to be trained & implemented in the contract research and testing lab (CRTO) / contract research and manufacturing organization (CRMO). • GLP audits should be performed frequently for Vendors and CRTO / CRMO sites. • We can learn GLP from customer / MNCs and external training programs. • We can learn GLP from 483’s, warning letters and Non-Compliance reports of other companies. • Perform GAP analysis to identify gaps in the system and correct and implement GLP.
  119. 119. A to Z of GLP
  120. 120. • There is no end for discussion on Good Laboratory Practices. • Good Laboratory Practices are to be improved continuously to determine the exact quality of the product. • GLP is required to be followed to ensure Compliance and Data Integrity (http://www.slideshare.net/skvemula/presentation-on-data-integrity-in-pharmaceutical- industry) and proper guidance on usage of computers and Chromatographic Data Systems (http://www.slideshare.net/skvemula/chromatography-data-system-cds-in-pharma). • GLP helps to understand the importance of each step to minimize errors (http://www.slideshare.net/skvemula/understand-the-importance-of-each-step-to-minimise- laboratory-errors) during the analysis and to reduce Laboratory errors (http://www.slideshare.net/skvemula/laboratory-errors).
  121. 121. • GLP helps to follow best practices and precautions to produce accurate results with the knowledge of Sample preparation techniques (http://www.slideshare.net/skvemula/sample-preparation-techniques-of-solid-dosage-forms) and to follow Good Chromatographic practices (http://www.slideshare.net/skvemula/good-chromatographic-practices) and guidance on proper Peak integration (http://www.slideshare.net/skvemula/hplc-peak-integration-for-chromatography- 38032765). • GLP is also required to investigate the atypical results (http://www.slideshare.net/skvemula/investigating-aberrant-potency-values-in-pharma-analysis) and to ensure that only quality product should reach to the patient.
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