Industries: Pharma / Biotech

Lyophilization Technology – Practical Application of the Scientific Principles

Course Director: Edward W. Sunbery, R.Ph.

Course Description - Course runs 9:00 to 5:00 both days

This course is designed to provide attendees with a practical approach to understanding the scientific and engineering principles of lyophilization. The topics discussed during the course will be presented to explain the physical, physicochemical, biochemical, and engineering principles of the science and engineering behind modern freeze-drying. The practicality of the course allows the attendees to get a firm understanding of the theoretical applications of freeze-drying and how these can be easily used and applied to their day to day lyophilization projects.

Topics covered will include:

  • History of lyophilization
  • Properties of water and ice
  • Supercooling and supersaturation
  • Phase behavior during freezing
  • Thermal characterization
  • Annealing
  • Visco-elastic properties of amorphous phases
  • Collapse phenomena
  • Vacuum physics
  • Product formulation
  • Heat and mass transfer
  • Cycle development
  • End point determination
  • Residual moisture
  • Process scale-up and sonic water vapor flow
  • IQ/OQ/PQ
  • Process control and optimization
  • Fault finding and problem solving of systems and formulations
  • Freeze dryer mechanical and control systems
  • Recipe development
  • Following the process

Who Should Attend

This two-day lyophilization training course is designed for those in the pharmaceutical, biomedical, medical device/diagnostic, and biotechnology industries who develop and/or produce lyophilized (freeze-dried) products. This includes, but is not limited to, professionals in Quality Control, Quality Assurance, Validation, Pharmacists, Managers, Microbiologists, R&D Groups, Biochemists, Pilot Plant Operators, Chemical Engineers, Production Supervisors, Chemists, Equipment Maintenance, and Mechanical Engineers. The course will also benefit those in other departments who find lyophilization among their responsibilities.

First Day

Module 1: Introduction

  • Harris and Shakell – Vacuum Desiccation of Rabies Virus
  • Changes in the State of Water
  • Molecular Structure
    • Ionic Bonds
    • Covalent Bonds
    • Hydrogen Bonds
    • Van der Waals Force
  • Water and Transient Hydrogen Bonds
    • Molecular Distance and Kinetic Energy
  • Molecular Structure of Liquid Water and Solid Ice
  • Molecular Structure of Liquid water and Gas Vapor
  • Sensible Heat Versus Latent Heat
  • Heating Curve of Water
    • From Below 0ºC to Above 100ºC
  • Heating Character of Water and Volume
  • Barometric Pressure
  • Barometric Pressure and Altitude
  • Vapor Pressure
  • Water Vapor – Temperature and Pressure
  • Dew Point
  • Frost Point
  • Ice / Water / Vapor - Phase Diagram – Temperature and Pressure
  • Colligative Properties of Water and Chemical Potential
    • Solubility
      • Polar Versus Non-Polar
    • Vapor Pressure
    • Boiling Point
    • Freezing Point
    • Osmotic Pressure
  • Other Issues
    • Glass Transition Temperature
  • Polymorphism

Module 2: Solidification

  • Challenge – Converting Solutes in Water to Solutes in Ice
    • Temperature – Regulate Molecular Motion and Attraction
    • Chemical Potential – Impacts Freezing Point Depression
    • Glass Transition – Unique to Polymeric Molecules
  • Solid Phase Transition and Pressure
  • Solid Phase Transition and Solutes
  • Solidification of Water
    • Locking Hydrogen Bonds
    • Nucleation – Molecular Clusters
      • Delayed Nucleation
    • Crystal Morphology and Cooling Rates
  • Solidification and Density (Water Floats)
  • Solidification Patterns and Fill Depth
  • Solidification of Water and Chemical Potential
  • Electrolytes and Freezing Point Depression
  • Salt is Water Exhibits Multiple Freezing Points
  • Non-Electrolytes and Freezing Point Depression
  • Solidification of Water and Electrical Conductivity and Resistance
  • Solidification and Glass Transition Temperature
    • Low Temperature Differential Scanning Calorimetry
  • Cooling Rates
    • Nucleation and Crystal Growth
    • Uncontrolled Nucleation
  • Other Process Variables
    • Temperature
      • Shelf Temperature Equilibrium Time During Start Up
      • Product Temperature and Distribution Equilibrium During Loading
    • Trays – Limit Direct Contact Between Containers and Shelf
    • Tubing Versus Molded Vials
  • Nucleation Control
    • Ice Fog
    • Depressurization
    • Ultra Sonic Vibration
    • Container Modification
  • Crystalline Material in Solution
    • Freezing Temperature
  • Complex Solutions and Colloidal Suspensions
    • Collapse Temperature
      • Hot and Cold Stage Microscopy
    • Cellular Material and Protein Formulations
      • Super Cooling
      • Spray Freezing
    • Crystal Size Control
      • Annealing
    • Spray Freezing and Particle Size Control

Module 3: Sublimation (Primary Drying)

  • Challenge – Convert Ice to Vapor and Back to Ice with no Liquid Phase
  • Collapse Temperature
  • First Law of Thermodynamics
    • Energy, Heat & Work
  • Second Law of Thermodynamics
    • Entropy & Efficiency
  • Third Law of Thermodynamics
    • Motion & Temperature
  • Water Boiling Point Temperature and Pressure
  • Vapor Pressure and a Materials Ability for Phase Change
  • Changes in the Phase of Water and Volume
  • Gas Laws
    • Charles Law
    • Boyles Law
  • Chamber Capacity and Design
  • Condenser Capacity and Design
  • Condenser Location
  • Phase Transition Control and Vapor Flow
    • Choke Point
    • Maximum Sublimation Rate
    • Dew Point
    • Frost Point
      • Vapor Pressure Over Ice
        • Insulation Factor for Ice
      • Sublimation Variables Heat Transfer
        • Convection
        • Radiation
        • Conduction
      • Other Sublimation Variables
        • Container Product Fill Depth
        • Loading Trays and Rings
        • Container Design
        • Temperature Sensors
        • Freeze Dryer Design
        • Container Location in Freeze Dryer
        • Partial Stopper Placement
      • Sublimation End Point Determination
        • Visual Observation
        • Temperature
          • Product
          • Thermocouple
          • Thermocouple Gauge
        • Pressure
          • Capacitance Manometer – Absolute Pressure
          • Pirani Gauge – Relative Pressure
          • Pressure Differential
          • Pressure Rise
        • Vapor
          • Relative Humidity
          • Dew Point
          • Tunable Diode Laser Absorption Spectroscopy
            • Concentration
            • Temperature
            • Velocity
          • Freezing and Sublimation Summary
          • Sublimation Versus Desorption

Module 4: Desorption (Secondary Drying)

  • Challenge – Reduce Bound Water Following Sublimation of Free Water
  • Bound Water
    • Salt Concentrates with Freezing and Stops Ice Crystal Formation
    • Free Water Binds to Charged Sites
  • Review
    • Molecular Bonding Forces
    • Intermolecular Forces
    • Hydrogen Bonds
  • Objective – Residual Moisture for Optimal Stability
    • Biological (Amorphous) Material
    • Crystalline Material
  • Secondary Drying
    • Desorption – Temperature and Pressure
    • Desorption- Product Degradation
    • Mechanism of Action
    • Efficiency
    • Drying Rates
      • Crystalline Material
      • Amorphous Material
    • End Point Determination
      • Sample Location
      • Sampling Procedure
      • Sample Analysis
    • Rationale for Final Moisture Level
    • Analytical Moisture Determination Methodology
      • Thermal Gravimetric Analysis
      • Karl Fischer
      • Combination Methods
        • Thermal Gravimetric Analysis / Gas Chromatographic Mass Spectroscopy
        • Fourier Transformer Infrared Spectroscopy
        • Near Infrared Spectroscopy (NIR)
      • Potential Chemical Routes of Degradation
        • Hydrolysis
        • Redox (reduction – oxidation)
        • Catalysis (metal)
        • Photo Catalysis (light)
        • Temperature
      • Routes of Degradation for Vaccines
      • Microbiological Issues
        • Water Activity (aw)
        • Growth Activity
      • Kinetics and Rate Process Evaluation
      • Container Closure Insertion and Sealing
      • Inspection

V Formulation and Raw Materials

  • Available Data Review
    • Material Characterization
    • Routes of Degradation
    • Excipient Characteristics
    • Solution Characteristics
    • Process Conditions
    • Component Compatibility
  • Compare
    • Similar Chemical Structure & Associated Formulation
    • Similar Therapeutic Categories
  • Product Development and Regulatory Rationale
    • Purity – 1906 Food & Drug Act
    • Safety – 1938 Federal Food Drug and Cosmetic Act
    • Efficacy – 1962 Kefauver – Harris Amendments
      • Optical Activity
    • Process Validation – Expectations
    • Basic Excipient Selection Issues
      • Buffers (pH)
      • Osmotic Pressure
      • Bulking Agents & Annealing
      • Cryo-Protectants
      • Surfactants
      • Co-Solvents
      • Miscellaneous Agents
    • Sample Formulations
      • Indomethacin for Reconstitution
      • Ethacrynate Sodium for Reconstitution
      • Pantoprazole Sodium for Reconstitution
      • Varicella Virus Vaccine, Live
      • Alteplase
      • Teduglutidum for Reconstitution
      • Raw Material Review

Module 5: Unit Operations

  • System Components (primary, support systems & sensors)
  • Freeze Dryer Unit Operations
    • Condenser Defrost
    • Clean in Place
    • Steam in Place
    • Leak Test
    • Bulk Solution and Aseptic Filtration
    • Subdivision
    • Chamber Loading and Freezing
    • Freezing – Solidification
    • Primary Drying (sublimation)
    • Secondary Drying (desorption)
    • Vial Stopper Insertion
    • Sealing Stoppered Vials
    • Inspection – Acceptance Criteria

Second Day

Module 6: Process Trouble Shooting

  • Study 1 – Vials Exhibiting Halos
    • Visual Observations
      • Review: Surface Tension
    • Investigation - 1
      • Issue: Equipment Set Up
      • Acceptance Criteria
    • Investigation - 2
      • Issue: Filling Line Maintenance and Operation
      • Acceptance Criteria
    • Investigation - 3
      • Issue: Subdivision and Freezing Holding Time
      • Acceptance Criteria
  • Study 2 - Broken Vials
    • Visual Observation
      • Review: Freezing and Crystallization
    • Investigation -1
      • Issue: Container Diameter, Thickness and Fill Depth
      • Acceptance Criteria
    • Investigation – 2
      • Issue: Solidification Procedures (Time & Temperature)
      • Acceptance Criteria
    • Investigation – 3
      • Issue: Sample Dimensions
      • Acceptance Criteria
  • Study – 3 Lyophilized Cake Shrinkage
    • Visual Observation
      • Review: Drying Parameters
    • Investigation – 1
      • Issue: Container Specifications
      • Acceptance Criteria
    • Investigation – 2
      • Issue: Closure Placement
      • Acceptance Criteria
    • Investigation – 3
      • Issue: Vendors Raw Material Specifications
      • Acceptance Criteria
    • Investigation – 4
      • Issue: Primary Drying Procedures
      • Acceptance Criteria
  • Study – 4 Product Frothing
    • Observation
      • Review: Primary Drying Mechanism of Action
    • Issue: Temperature and Pressure Control
    • Acceptance Criteria
  • Study – 5 Product Pulling Away from the Container
    • Visual Observation
      • Review: Temperature and product Freezing
    • Investigation – 1
      • Temperature: Record vs. Actual Conditions
      • Acceptance Criteria
  • Study 6 – Out of Specification Moisture
    • Analytical Observation
      • Review Qualification and Validation Parameters
    • Investigation – 1
      • Equipment and Raw Materials
      • Acceptance Criteria
    • Investigation – 2
      • Validated Procedures
      • Acceptance Criteria
    • Investigation – 3
      • Analytical Laboratory Practices
      • Acceptance Criteria
    • Investigation – 4
      • Secondary Process – Steam Sterilization
      • Acceptance Criteria
    • Investigation – 5
      • Secondary Process – Closure Washing
      • Acceptance Criteria
    • Background Support Information
      • Closure Manufacturing
        • Calendaring
        • Molding
        • Curing
          • Impact from Hot and Cold
  • Study 7 – Analysis of Complete Collapse
    • Visual and Process Documentation Observations
      • Data Summary
    • Study -1 Recorded Freezing Data Analysis
    • Study – 2: Recorded Primary Drying Data Analysis
    • Study – 3 Recorded Secondary Drying Data Analysis
    • Data Summary
      • Complete Collapse and Melt Back
      • Acceptance Criteria
  • Study 8 – Analysis of Partial Collapse
    • Visual and Process Documentation Observations
      • Data Summary
    • Study -1 Recorded Freezing Data Analysis
    • Study – 2: Recorded Primary Drying Data Analysis
    • Study – 3 Recorded Secondary Drying Data Analysis
    • Data Summary
      • Partial Collapse and Melt Back
      • Acceptance Criteria
  • Case Study Review – Freezing & Drying – Complete vs. Partial Collapse
  • Lyophilization
    • Process Summary
      • Freezing
      • Sublimation
      • Desorption
    • Acceptance Criteria
      • Prepare and Examine Plotted Process Data
        • Confirm No Process Excursions Beyond Validated Parameters
        • Confirm Temperature and Pressure Control Parameters
      • Trouble Shooting Questions
        • What is the Sublimation Rate and Collapse Temperature?
        • How is the End of Primary Drying Determined?
        • What is the Analytical Support for the End of Secondary Drying?
        • What is the Impact on Vapor Flow from Closure Design and Placement?

Module 7: Equipment Design and Unit Operations

  • Equipment Design
    • Manifold Freeze Dryers Design
      • Sample Preparation
      • Sequence of Operation
      • History of Manifold Freeze Drying
    • Shelf Dryer Design
      • Cylindrical Chambers
        • Internal Condenser and Locations
        • External Condenser and Locations
      • Cubical Chambers
        • External Condenser Locations
      • Aseptic Production Equipment Design
        • Operational Advantages
      • Isolation Valve Design
        • Butterfly vs. Poppet Valve
      • Sample Thief Design
    • Unit Operations – System Component Summary
      • Condenser Defrosting
      • Clean in Place
      • Steam in Place
      • System Leak Test
      • Product Subdivision
      • Chamber Loading
        • Product Solidification
        • Primary Drying
        • Secondary Drying
      • Container Closure Insertion
      • Sealing Stoppered Containers
    • Design Issues – Support Systems
      • Pressure
        • Pump Design
          • Rotary Vane Pump
          • Mechanical Blower
          • Reciprocating Pump
        • Vacuum Pump Trouble Shooting Issues
          • Failure to Start
          • Difficult to Start
          • Very Noisy Operation
          • Failure to Meet Pressure Set Point
          • Failure to Control >+ 10% of the Pressure Set Point
        • Temperature
          • Single Refrigeration System Components
            • Compressor
            • Condenser
            • Expansion valve
            • Evaporator
          • Multi Stage Refrigeration System Components
            • Heat Exchanger
          • Refrigeration System Trouble Shooting Issues
            • Compressor Fails to Start
            • Compressor Starts – No Cooling to Shelf or Condenser
            • Lack of Uniform Shelf Temperature
            • Poor Control Around Shelf Temperature Set Point
            • Condenser Temperature – Set Point Control >+ 10ºC
          • Design Issues – Instrumentation
            • Temperature
              • Thermocouple
                • Parallel vs. Series Circuits
                • Equipment Selection Issues
                  • Temperature vs. Change in Electromotive Force
                  • Temperature Range
                  • Mechanical and Thermoelectric Properties
                  • Chemical Stability and Resistance to Reactivity
                • Trouble Shooting Thermocouple Issues
                  • Loss of Calibration
                  • Signal Noise
                  • Thermal Shunting
                • Pressure
                  • Phase Transition of Water and Volume
                  • Barometric Pressure
                    • Height of Mercury in a Semi-Sealed Tube
                  • Instrumentation – Direct Pressure Reading
                    • McCloud Gauge
                    • Capacitance Manometer
                  • Instrumentation – Indirect Pressure Reading Based on Vapor Cooling
                    • Thermocouple Gauge
                    • Pirani Gauge
                  • Vapor
                    • Instrumentation – Based on a Stored Electrical Charge
                      • Dew Point Monitor
                    • Instrumentation – Based on Spectroscopy
                      • Tunable Diode Laser Absorption Spectroscopy
                        • Concentration
                        • Temperature
                        • Velocity
                      • Design Issues – Container Closure Stoppering
                        • Equipment Design
                        • Container Stopper Procedure
                        • Trouble Shooting
                      • Aseptic Air Vent Filter Design
                        • Bubble Point Procedure and Acceptance Criteria

Module 8: Equipment Capability

  • Freeze Drying Challenges
    • Barriers to Entry
    • System Complexity
    • Diluents
  • System Components
    • Primary Components
    • Support Systems
    • Sensors
  • Validation Expectations
  • Validation – Life Cycle
  • Facility – Installation
    • Scope
      • Environmental Controls
      • Electrical Supply
      • Water Resources
    • Equipment - Installation Capability
      • Purpose
        • Acceptance Criteria
        • Documented Evidence
      • Scope
        • Documentation
        • Materials of Construction
        • Operational Specifications
      • Equipment – Operational Capability, Procedures, Acceptance Criteria and Trouble Shooting
        • Chamber Temperature
          • Maximum Heating and Cooling Rates
          • Sublimation and Deposition Rates
        • Chamber and Condenser Pressure
          • Maximum Vacuum Pump Pressure
          • Low Pressure Integrity Test
          • High Pressure Integrity Test
          • Pressure Set Point Control During Drying
        • Condenser Temperature
          • Condenser Cooling Rate and Maximum Temperature
        • Vapor Valve
          • Choke Point
            • Procedure and Acceptance Criteria
          • Maximum Sublimation Rate
            • Procedure and Acceptance Criteria
          • Container Closure Stoppering
            • Acceptance Criteria

Module 9: Process Capability

  • Process Validation Expectations
  • Critical Product Parameters
    • Morphology
      • Crystalline Versus Amorphous Material
    • Temperature Analysis
      • Glass Transition Temperature
      • Collapse Temperature
    • Dry Cake Characteristics
      • Color
      • Density
      • Dose Uniformity and Purity
      • Residual Moisture Range
    • Constituted Product Attributes
      • Concentration, Purity, pH, Color and Odor
      • Rate of Solution (Time to Reconstitute)
      • Degree of Solution (Product Clarity)
    • Critical Process Parameters
    • Independent Parameters
      • Shelf Temperature
      • Chamber Pressure
      • Time
    • Dependent Parameters
      • Product Temperature
      • Condenser Temperature
    • Critical Data
      • Product Temperature Range at Critical Process Time
      • Chamber Location and the Impact on Product Attributes
      • Identify Extreme and Representative Chamber Locations
    • Critical Process Parameter Procedures
    • Product Temperature Uniformity Independent of Chamber Location
    • Loading Studies
    • Freezing Studies
    • Primary Drying Studies
    • Secondary Drying Studies
    • Boundary Studies
      • Alert and Action Levels
    • Supporting Stability Data
    • Degradation – Mechanism of Action
      • Thermal Stress
      • Freeze-Thaw Stress
      • Mechanical Stress
      • Light Stress
      • Oxidation Stress

Module 10: History

  • Early 1900’s - Harris & Shakell -  Vacuum Desiccation of Rabies Virus
  • Rotary Vane Vacuum Pump
  • 1927 – First Patent Reference to Low Pressure Drying
  • 1933 – Clinical Studies with Freeze Dried Blood Plasma
  • 1940 - 1945 – 10,000,000 Units of Freeze Dried Plasma Produced for WWII
  • 1947 – Freeze Dried Orange Juice
  • 1948 – First Book on Large Scale Freeze Drying
  • 1951 – Basic Freeze Drying Parameters Defined
  • 1957 – Accelerated Freeze Drying
  • 1958 – Freeze Dried Milk
  • 1967 – Freeze Dried Rocket Propellant
  • Freeze Dried Taxidermy
  • Freeze Dried Flowers

Wet Document Recovery by Freeze Drying

Learning Objectives

Upon completion of this course, attendees will have a clear understanding of both the scientific and practical principles behind the formulation and development of lyophilized products. Additionally, participants will gain a firm understanding of lyophilization equipment including system operation, system components, system maintenance, system troubleshooting, IQ/OQ, and system validation. Attendees will also gain practical experience in applying what they have learned by working through several case studies in failed products and cycles. This course is presented in a very practical manner, so attendees will be able to quickly apply what they have learned to their lyophilization projects.

Testimonials

"The Course Director had deep knowledge of the field. He shared good examples and the course material was a great starting point to launch discussions relevant to our specific needs. He was very responsive and the material is useful for both training and for reference." Dom D.,(Ph.D.) Abbott Moleculars
"The lyophilization training course has great technical and very informative material to understand the science behind lyophilization process and the potential problems that may occur during production." Ella H., QA Manager, Gilead Sciences
"The Course Director was very knowledgeable and his real life experiences made the course that much better. He knows what he is talking about!" Kevin D., Director of Manufacturing, J&J
"Since I don't operate the machines or design product cycles I came to this course not knowing what to expect. The result of the course and discussions with instructors and attendees has given me the inspiration to become involved with the freeze-drying processes at my company in the future. The discussions not only gave technical information but brought the processes to my level of interest." Dan M., Bioreagents Technologist, Instrumentation Laboratory
"This was a very good review/introduction to lyophilization. I didn't have much background in any of this when I arrived, but now I feel like I can begin to ask the right questions and to focus on our particular application of freeze drying." John C., Bioreagent Technologist, Instrumentation Laboratory