© Institute for International Research, Inc All rights reserved. Module 2: Regulations

2 international Module 2 Purpose and Objectives  Module Purpose:  Content will presuppose a sterile lyophile in glass vials and will review general regulatory requirements.  Module Objectives:  After this module, you will be able to  Identify essential testing requirements  Understand what makes a machine comply with cGMP sterility and cleaning  Participate in a discussion of event recovery

3 international End Testing Requirements  USP Injections  Uniformity of Dosage Units  Bacterial Endotoxin  Water Determination  Reconstitution time  Identification & Assay  Elegance?  Stability

4 international USP  Injectable Finished Product Requirements  Product must pass USP sterility test  There will probably be an endotoxin specification  Reconstituted product must pass a visible particle inspection  Reconstituted product must pass a Small Volume parenteral particulate test.

5 international Uniformity of Dosage Units  Tier Testing 1:  No more than 1 of the 10 dosage units lies outside the range of 75% to 125% of the label claim and none of the 10 doses is outside the range of 65% to 135% of the label claim. If 2-3 units are outside 75% to 125% of the label claim, but no unit outside 65% to 135% of the label claim, test 20 additional units.

6 international Uniformity of Dosage Units  Tier Testing 2 (on 20 additional units)  No more than 3 of 30 dosage units are outside the range of 75% to 125% of the label claim and none of the 30 dosage units is outside the range of 65% to 135% of the label claim.

7 international Bacterial Endotoxin  The general requirement for injectables is less than or equal to 0.25 EU/mL of reconstituted product. This becomes a problem if the reconstitution volume is less than the fill volume for lyophilization.  Some drugs may have a specific limit  Note: ophthalmics and intrathecals will have a lower limit.

8 international Moisture Determination  Sulfur Dioxide Titration I 2 + SO 2 +2H 2 O2HI + H 2 SO 4 In Aqueous Solution, this reaction works very well to titrate sulfur dioxide and it is overwhelmingly shifted to the right by the enormous concentration of water. Karl Fischer figured out that he could titrate water in a non-aqueous solvent and pull the equilibrium to the right by using a neutralizing base to remove the H 2 SO 4.

9 international Karl Fischer Reaction CH 3 OH + SO 2 + RN ==> [RNH]SO 3 CH 3 H2OH2OI2I2 RN [RNH]SO 4 CH 3 + 2[RNH]I RN = Pyridine pH = 5 to 7

10 international Karl Fischer Reactions  Volumetric KF – Measures the volume or weight of added titrant.  Coulometric KF – measures the current generated during the titration.

11 international Reconstitution Time  Must be measured by a standard procedure  Solution should be determined versus a standard.

12 international Other Tests  Assay  Elegance  Stability – Moisture instability due to stopper adsorption of water

13 international Sterility – Filtered Gasses

14 international Dead Legs

15 international Chamber penetrations  Door including Window(s) & Door Ports  Isolation valve (path to condenser)  Capacitance Manometer 0 – 2 torr  Pressure (full vacuum to 15+ psi)  CIP/Steam/Gas inlet  Thermocouple inlet  Stoppering Ram inlet  Validation Port(s)  Shelf coolant Inlet & Exit  Drain

16 international

17 international Leak Testing  PDA specification is 0.02 mbar∙liter/sec  0.02mbar = 15 mtorr  Example:  If a system (chamber + condenser) has an internal volume of 2267 liter, then it should have a pressure rise of less than

18 international Volume Determination

19 international Volume and Leak Test Data Note the potential for large error in the calculation if the known volume and pressure aren’t accurate. Atmospheric pressure varies from day to day.

20 international Leak Rate Calculation  Once the volume is known, the leak rate can be calculated. Each lyophilizer will have a different Leak Specification (in mtorr/min or mtorr/hr) depending on its volume.

21 international Normal Cycle Variance  Freezing – everything must be solid.  Target is usually -50  C. Range is -40 to  Vacuum – Variation in a dry chamber should be slight. E.g. 100mtorr +1mtorr  Variation during a run will depend on how well the PID loop is tuned and what software control scheme is used.  100mtorr + 25 mtorr may be seen. If so, it should be quick – full cycle every 5 min.

22 international Normal Cycle Variance  Condensers: Within a small time frame (10 minutes) variance may be + 2  C.  Drift during a run is common. Often, there is no set point. The condensers are run as cold as possible using the available compressor power. Drift is caused by shunting coolant to the shelf package.

23 international Normal Cycle Variance  Shelves: A dry shelf specification is often + 1  C. When product is on the shelves, the ability to hold a temperature will depend on the available coolant. Most users will continue to demand + 1  C from setpoint.

24 international Shelf Temperature Under Load Red = SetPoint Blue = Shelf In Green = Shelf Out

25 international Cleaning  Use CIP if available. Otherwise, do a thorough manual cleaning.  Check for broken glass.  Validate with Riboflavin and a black light.  Test with Swabs followed by TOC.  Clean to background – about 500 ppb  Sterilization is not substitute for cleaning.

26 international Issues  An unending list.  Power Outage  Software error  Chamber overload  Operator error  Equipment failure (long sublist)  Sensor failure  Data recording failure  Investigate to assure non-recurrence.  Use good science.

27 international Exercise 2.1: Valve 1Valve 2Valve 3Valve 4Valve 5Valve 6Valve 7Valve 8Valve 9 Heating of the Filter Housing Steam Sterilization of Filter Interim Drying of Filter Media Fill Housing with Water for Testing Perform Water Intrusion Integrity Test Drain Filter Housing Dry Filter Media Valve for Nitrogen Pressure Control

28 international Module 2 Quiz  Participant Directions:  Divide into pairs  Take 5 minutes to complete quiz  Correct answers will be reviewed as a large group during debrief

29 international Question 1 of 3  Quiz Question: 1.A ‘dead leg’ is a closed end pipe that is how many pipe diameters away from the vessel? a.3 pipe diameters b. 6 pipe diameters c.9 pipe diameters

30 international Question 2 of 3  Quiz Question: 2.When does USP Particulate Matter in Injections apply? a.Always on a sample after reconstitution. b.Whenever a sample >100 mL is reconstituted with sterile liquid. c.Always, if reconstituted with sterile liquid. d.Whenever a specific monograph requires the test. e.Answers ‘b’ and ‘d’ f.None of the above

31 international Question 3 of 3  Quiz Question: 3.Suppose a product has failed to advance out of freezing. i.e. The product froze correctly, but the condensers never reached set-point and thus vacuum was not initiated and primary did not start. What to do?

32 international Module 2 Summary  Required Testing for a Pharmaceutical Lyophilization Product.  Basic Pharmaceutical Equipment Requirements  Begin thinking about event recovery.