Sterile Injectable Preparations Quiz

Sterile Products Manufacture - Quality Evaluation

• Dr. Laura Urbano, F154 Hillside House, [email protected], 01707 281372
• Quality evaluation of sterile products is covered

Learning Objectives

• Critically understand the quality requirements for sterile products and compare them to pharmacopeial standards
• Understand the sources of particulates and endotoxins in sterile manufacturing
• Review the rationale, techniques, and acceptable limits for quality tests relating to sterile products

Sterile Preparations

• General monograph in BP (Ph. Eur. monograph 0520) defining sterile preparations for injection, infusion, or implantation in human/animal bodies
• Excipients, containers, preservatives, and water for injections are included in the definition

Quality Tests for Compliance

• Tests for particulate contamination (sub-visible particles, 2.9.19) -covered in a workshop
• Test for sterility (2.6.1)
• Consistency of formulated preparations (Appendix XII C)
• Uniformity of dosage units (2.9.40)
• Uniformity of content (2.9.6)
• Extractable volume of parenteral preparations
• Tests for bacterial endotoxins (2.6.14) -Tests for pyrogens (2.6.8)

Types and Sources of Particulate Matters in Injectable Preparations

• Table listing sources (intrinsic/extrinsic) of particulate matters in injectable preparations including dust, fibers, biologics (e.g., insect parts, microorganisms, pollens), hair, fibers from anthropogenic origin, skin, paint/coating chips, metal, rust, minerals, polymers, glass, extraneous material (e.g., from rubber stopper components)
• Additional table providing intrinsic/extrinsic classification of particulate matters from packaging materials and solution/Formulation components, including rubber, silicone, glass, polymers, precipitates, oligomers, degradants, agglomerates, undissolved material, glass lamellae, silica, rubber, plastic, metal, and filter/consumables.

Summary of Available Data

• Data summary for sterile injection applications submitted to USFDA between September 2010 and July 2011, including dosage route, container type, container volume, subvisible particle limit, method, 210 µm particle average ± SD, 225 µm particle average± SD, visible particle results (stability batches) and comments regarding conformity.

Techniques for Sub-Visible Particles

• Light Obscuration Particle Count Test: Apparatus measures light blockage to determine particle size and numbers; calibration with spherical particles (10 and 25 µm); particle-free water as a dispersion medium; laminar-flow cabinet for a particle-free environment, determination of 5 samples for particulate contamination, and environment should be suitable for particle counting.
• Microscopic Particle Count Test: Suitable binocular microscope and filter assembly used to retain particulate contamination; Membrane filter is used for examination. Other requirements are the same as in the Light Obscuration Particle Count Test method.

Criteria for Acceptability

• Criteria for solutions for infusion/injection (>100mL): average numbers <25 per mL (size≥10 µm) and <3 per mL (size ≥ 25 µm)
• Criteria for solutions for infusion/injection (≤100mL): average numbers <6000 per container (size ≥ 10 µm) and <600 per container (size ≥ 25 µm)

Consistency of Formulated Preparations

• Uniformity of weight: Procedures for measuring consistency of weight involve: removing labels, washing and drying containers; rinsing containers with water and then alcohol; drying at 100-105°C for 1 hour; weighing container contents, and measuring deviation from average mass.
• Uniformity of weight (mass): Standards related to deviation from the average mass, based on the pharmaceutical form (e.g., tablets, capsules, granules, powders, suppositories) are provided.
• Uniformity of content: Method involves chemical analysis of individual contents in a sample of 10+20 units for calculating percent content and deviation from the average content. Criteria includes all 10 units within the 85-115% of the average content; if any units outside 85-115% but are within 75-125% more units must be tested.

Extractable Volume of Parenteral Preparations

• Single Dose Containers: Procedures for measuring extractable volumes and criteria for volume measurements using syringes and cylinders.
• Multi Dose Containers: Similar procedures and criteria apply for multi-dose containers, including measuring extractable volume.
• Cartridges and Pre-filled Syringes: Same process as single dose containers
• Infusions: Volume be measured for determining occupancies at least 40% of cylinder.

Endotoxins

• Appendix XIV C: Test for Bacterial Endotoxins (LAL Test). Used to detect bacterial endotoxins from the outer membrane of gram-negative bacteria using amoebocyte lysate.
• Appendix XIV D: Test for Pyrogens. Detects pyrogens, i.e., heat-resistant lipopolysaccharides from gram-positive bacteria, mycobacteria, fungi, and viruses. The significance of endotoxins in the pharmaceutical industry is addressed. The bacteria that induce pyrogens and cause fever in humans are also discussed.

Pyrogens

• Prevention is key for pyrogens removal.
• Primary source is water.
• Pyrogens are destroyed by distillation process, but also exist in water used for cleaning containers & closures.
• Sterilization methods using dry heat (glass) and washing with pyrogen-free water (rubber closures).
• Chemical raw materials used in parenteral formulations should be crystallized using pyrogen-free water or other solvents.

Endotoxin Methods

• LAL tests (Appendix XIV C): Depyrogenate all glassware at 250°C for 30 minutes before the test. Components of LAL test are listed, including reagents & solvents.
• Procedure for dissolving product in water for bacteria endotoxin test. pH must be adjusted to 6-8
• Endotoxin Detection Methods: Gel-clot, turbidimetric, chromogenic methods are detailed.

LAL Tests (Appendix XIV C) - Maximum Valid Dilution

• Describes the maximum dilution (MVD) allowed for determining endotoxins in a sample, correlating it to the endotoxin limit and concentration of the test solution (λ).
• Defined A as labelled lysate sensitivity or lowest concentration in a calibration curve, and K/M as the endotoxin limit where K represents the threshold pyrogenic dose per kg of body mass, and M is the maximum recommended dose per kg of body mass.

Endotoxin Limits

• Lists endotoxin limits (K values, in IU) based on different routes of administrations (Intravenous, Intravenous for radiopharmaceuticals, Intrathecal, Parenteral).

Pyrogen Testing

• Appendix XIV D: Describes the selection criteria for rabbits and the preparation of the test animals (age, sex, weight, diet).
• Material requirements include glassware, syringes, and needles, along with the sterilization process.
• Describes pyrogen tests, including temperature recording before and after injection; pre-test measurement of temperature after pyrogen-free NaCl injection; injection of test substances; sample preparation (0.5–10mL/kg body weight) and measurement of body temperature over 3 hours.
• Lists the criteria for passing (temperature variation < 1.15 °C) and failing (>2.65°C), and what to do if sum of temperature increases is between 1.15 and 2.65°C.

Pyrogen Testing - Different Methods

• Rabbit Pyrogen Test (RPT): Measures the raise in body temperature after injection of a drug. In-vivo testing.
• Monocyte Activation Test (MAT): Detects pyrogens by measuring cytokines released by blood monocytes. In-vitro testing.

References & Resources

• Aulton's Pharmaceutics (2013)
• Langille (2013) study on particulate matter in injectable drug products
• Current BP and USP standards

Test your knowledge on the standards and quality control for sterile injectable preparations. This quiz covers topics such as particulate matter, quality tests, and specifications for injectable pharmaceuticals. Perfect for students and professionals in pharmaceutical sciences.