1 Why Do I Test, What Do I Test & When Do I Test It? Ross Caputo, PhD Chief Technical Officer Eagle Analytical Services
2 Disclosures I, Ross Caputo, declare no conflicts of interest, real or apparent, and no financial interests in any company, product, or service mentioned in this program, including grants, employment, gifts, stock holdings, and honoraria.
3 Learning Objectives At the conclusion of this program, the participating pharmacist or pharmacy technician will be able to: Discuss why science-based testing must be performed and determine when preparations require testing Name available testing methods for chemical and biological attributes of sterile and non-sterile compounds Identify how to develop a science-based testing program as part of a pharmacy's Quality System
4 WHY TEST? Why is testing compounds so important?
5 Why Test? USP and FDA guidelines mandate specific testing requirements for compounded preparations. These science-based guidelines have been established to ensure medication quality and patient safety. These guidelines are not just suggestions, they are considered enforceable by the FDA!!!
6 Testing must be performed: Why Test? To validate compounding processes To validate compounding personnel To establish extended beyond-use-dating To verify that a compound falls within labeled specifications To identify microorganisms captured during environmental monitoring As part of the Quality Program required in USP <795> and <797>
7 Where to Test? Compounders are responsible for vetting the laboratories they use for testing. Choose a laboratory that is: FDA registered DEA registered ISO Accredited
8 WHEN TO TEST When should compounds be tested?
9 When to Test Testing for Release Validate safety & quality of a compound Comply with USP, ISO, & FDA regulations Stability Studies Scientifically justify extended BUD Observe stability of preparation over time Determine storage conditions Environmental Sampling Identify microbes Determine corrective actions
10 When: Testing for Release To comply with guidelines set by regulatory agencies, compounders must ensure the quality, safety, and stability of compounded preparations. To accomplish this, preparations can be tested prior to release for: Sterility Bacterial endotoxins Potency Microbial enumeration
13 WHAT TO TEST Release Tests for Sterile Products
14 WHY: Sterility Test 503A facilities are required to sterility test CSPs that: Are produced in batches of 25+ units Exceed USP <797> beyond-use-dating guidelines 503B facilities are required to test every batch of sterile drug product prepared. Testing must follow a compendial (or better) methodology to support the claim that a product is sterile.
15 WHY: Sterility Test USP <71> sterility test Rapid ScanRDI sterility test <71> defines the compendial requirements for conducting sterility tests 14 day incubation period Utilizes fluorescent labeling and laser scanning to detect bacteria, mold, yeast & fungus 2 day turnaround time Sensitive enough to detect presence of a single cell
16 WHY: Method Suitability Testing Method Suitability Testing (MST) must be performed before sterility test results can be used to claim that a product is sterile. MST only needs to be completed once for any given formulation based on the largest batch size and highest concentration that will be produced. The MST should be performed by the same laboratory that will conduct the sterility tests.
17 WHY: USP <71> MST USP <71> MST consists of two components: Suitability test to confirm that the growth media supports the growth of certain microorganisms Validation test to demonstrate that no components of the preparation inhibit microbial growth
18 WHY: Rapid ScanRDI MST Rapid ScanRDI MST consists of two components: Confirms that formula & excipients do not cause interference with auto fluorescence of preparation Validates that components of formula do not inhibit uptake of fluorescent dye by viable organisms
19 WHY: USP <85> Endotoxin Test As bacterial endotoxins can pose health and safety hazards to patients, quantities of bacterial endotoxins cannot exceed threshold limits defined in USP <85>. USP <85> outlines testing requirements used to quantify the presence of bacterial endotoxins in CSPs.
20 WHY: Potency Testing Potency tests are designed to determine the concentration of an active drug in a sample. 21 CFR 211 requires laboratory determination of the identity and strength of each active ingredient prior to a preparation s release. USP <621> & <851> outline compendial methods for potency testing, such as using HPLC / UHPLC & Spectrographic instrumentation.
21 WHAT TO TEST Release Tests for Non-sterile Products
22 WHY: Microbial Enumeration Testing The presence of microorganisms in nonsterile preparations has the potential to: Reduce and / or inactivate therapeutic activity Adversely affect the health of patients Producers of compounded products are therefore required to ensure that nonsterile preparations have a low bioburden.
23 WHY: <61> Microbial Enumeration Testing Nonsterile products are required to comply with established specifications for microbiological quality as outlined in USP <61>. USP <61> microbial enumeration tests provide a quantitative evaluation of a product s microbial content to demonstrate compliance with established specifications.
24 WHY: <62> Tests for Specific Microorganisms Table 1 in USP <1111< sets acceptance criteria for the presence of microorganisms in a nonsterile preparation based on its route of administration. The presence of these objectionable pathogens & microorganisms cannot exceed the acceptance criteria. USP <62> testing is designed to demonstrate compliance with these requirements by quantifying the presence of specified microorganisms.
25 WHEN: Final Release Testing 503A 503B Test Non-Sterile Sterile Non-Sterile Sterile Closure integrity Initially in stability Initially in stability Initially in stability Initially in stability USP 71* sterility Every batch Every batch USP 85 endotoxin Every batch*** Every batch Potency ** Every dosage Initially every Every batch Every batch form every 6 months every technician, plus weight measure formula then annually Visual Every batch Every batch Every batch Every batch Particulate/pH Initially in stability Initially in stability Initially in stability Initially in stability
26 WHEN TO TEST Stability Studies
27 WHY: Stability Study Purposes of Stability Testing: Determine the beyond-use-date of a preparation Provide evidence on how environmental factors such as temperature, humidity, and light affect the quality of a preparation over time Establish appropriate storage requirements
28 WHY: Stability Study Stability studies test: The physical, chemical, biological & microbiological attributes of a preparation Preservative content & antimicrobial effectiveness Functionality (closure-container integrity testing, dose delivery system testing)
29 WHY: Stability Study 503A facilities Required to justify the beyond-use-date of compounded preparations whenever they exceed the BUD guidelines outlined in USP chapters 795 & 797 Required to test one lot 503B facilities Required to perform a stability over time study using a stability indicating method for all compounded formulations Required to test at least three lots
30 WHY: Stability Study USP <797> discourages compounders from using only publications to justify an extended BUD as this can only provide a theoretical BUD : Theoretically predicted beyond-use dating introduces varying degrees of assumptions and hence, a likelihood of error or at least inaccuracy (USP <797>) It should be recognized that the truly valid evidence of stability for predicting beyond-use dating can be obtained only through productspecific experimental studies (USP <797>)
34 WHY: Stability Indicating Assay All stability studies require the use of a stability indicating assay. A stability indicating assay: Determines the strength of active ingredients in a preparation Distinguishes active ingredients from their degradation products Allows a reliable estimation of the quantity of degradates in a preparation
35 WHY: Method Verification Method development is the process of developing stability indicating assays for any particular formulation. Method verification is required to demonstrate that the testing used in a study is accurate, appropriate, and will result in meaningful data. Method verification only needs to be completed once for any given formulation.
36 WHY: Method Verification Chapter 211 of the Code of Federal Regulations requires: Testing methods to be reliable, meaningful, and specific Verified under the actual conditions of use Documentation of the accuracy, sensibility, specificity, and reproducibility of test methods.
37 WHY: Sterility Testing As sterile products must remain free of microbial contamination throughout their entire shelf-life, sterility is considered to be a stability characteristic. Therefore, stability protocol should include confirmation of continuing sterility throughout the shelf-life. Minimally, sterility testing is performed at the start and the end points of a stability study.
38 WHY: <1207> Container-Closure Integrity Testing Sterility testing alone does not provide enough evidence of a product s continuing sterility due to practical & scientific limitations: Only detects microorganisms present at the time of the test Only detects microorganisms capable of growth in growth media used Destructive of samples tested As a result, container-closure integrity (CCI) testing is recommended to demonstrate the sterility of a preparation throughout its shelf-life.
39 WHY: <85> Endotoxin Test Sterile preparations must remain within bacterial endotoxin limits throughout a product s shelf-life. Bacterial endotoxin testing is performed at the initial time point and the end time point of a stability study.
40 WHY: Particulate Count Test As per USP <1>, parenteral injections and infusions must be essentially free of particulate matter. A USP <788> Particulate Count test is required to demonstrate compliance with subvisible particulate matter limits.
41 WHY: Particulate Count Test As per USP <788>, ophthalmic solutions must be essentially free of particulate matter. <788> requires a particulate count test for all ophthalmic solutions for which the monograph includes a test for particulate matter. Ophthalmic gels, emulsions, and suspensions are exempt from these requirements.
42 WHY: ph & Appearance USP <1163> recommends different quality assurance tests for stability protocols based on the preparation form. USP <1163> recommends ph measurements as per <791> as part of a stability study. ph measurements are indications of degradation and other product changes throughout the shelf-life. <1163> also recommends a visual inspection of the appearance of the product at each time point to inspect for any physical abnormalities, leaking, color changes, turbidity, viscosity, etc.
43 WHY: Microbial Enumeration Tests Compounders are required to ensure that nonsterile preparations have a low bioburden. USP <61> & <62> quantify the presence of microorganisms in nonsterile preparations.
44 WHAT TO TEST Environmental Sampling
45 WHY: Environmental Sampling Environmental Sampling (ES) should be one aspect of any well-developed quality management program. A robust ES program demonstrates that the sterile environment maintains acceptable viable and nonviable particle limits during its ongoing operations. An ES program seeks to detect microorganisms, potential hazards, and other adverse trends, allowing compounders to take corrective actions before a CSP becomes contaminated or another problem occurs.
46 WHY: Environmental Sampling An in-house ES program can consist of using media plates to take air and surface samples and monitoring the plates for the growth of colony forming units (cfus) as they incubate. Table 2 in USP <797> defines Action Levels based on the number and location of cfus identified. When Action Levels are exceeded, the pharmacy must initiate an investigation and develop a corrective action plan.
47 WHEN TO TEST Environmental Sampling
48 WHEN: Environmental Sampling USP <797> provides Action Levels & Alert Levels based on the numbers of cfus identified during environmental sampling. Even if Action Levels are not exceeded, <797> requires the microorganisms to be identified to ensure that there are no highly pathogenic (and potentially fatal) microorganisms in the sterile environment1. Any cfus recovered must be identified and a corrective action plan developed based on the type(s) of microorganism(s) recovered.
49 WHEN: Environmental Sampling USP <797> provides Action Levels & Alert Levels based on the numbers of cfus identified during environmental sampling. Even if Action Levels are not exceeded, <797> requires the microorganisms to be identified to ensure that there are no highly pathogenic (and potentially fatal) microorganisms in the sterile environment1. Any cfus recovered must be identified and a corrective action plan developed based on the type(s) of microorganism(s) recovered.
50 WHEN: Environmental Sampling HVAC non-viable all controlled area 503A 503B Test Microbiological Physical Microbiological Physical 6 month certification Monitor pressure, temp 24/7 6 month certification Monitor record temp/rh/pressure 24/7 Surface Air ISO 5 weekly Other ISO areas monthly Monthly 2 growth media ISO 5 every batch, Others daily ISO 5 every batch, Others daily Fingertip ISO 5 Every compounding day Every batch Media fills Each semi annually Each semi annually