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Container Closure Integrity – Dye Immersion & Bacterial Immersion

The Container Closure Integrity by Dye Immersion test is used to ascertain the integrity of a sample container. This test entails immersing the samples in a solution of methylene blue dye. The sample containers are tested with vacuum immersion and investigated for confirmation of leakage using an ultraviolet-visible (UV/Vis) or visual analysis.

The Container Closure Integrity test evaluates the adequacy of the closure in maintaining a sterile barrier. This test is performed on products such as filled vials, syringes, and devices. PDA Technical Report 27 recommends both a physical and microbial test when evaluating container closure integrity for pharmaceutical packaging. This test satisfies a worst-case scenario for the microbial recommendation and meets the requirements described in the FDA’s “Container Closure Systems for Packaging for Human Drugs and Biologics.” Nelson Labs performs this test using a unique chamber that allows us to apply pressure and/or vacuum to the challenge solution and test samples.

Applicable Standards

  • USP <1207>
  • PDA TR 27
  • FDA Docket 980-0021

Testing Locations

Study Outline

Dye Immersion: The Container Closure Integrity by Dye Immersion test is used to evaluate the closure on rigid containers. It can be used to test vials, screw cap bottles, and syringes. Samples are immersed in a dye solution and a vacuum is applied. The vacuum is released and the containers are washed to remove dye from the outer surfaces. The containers are then analyzed after exposure for the presence of dye using either an ultraviolet-visible (UV/Vis) or visual technique. The UV/Vis method has a higher sensitivity level for detecting the dye than the visual method.

Samples cannot have viscous or opaque fluids.

Bacterial Immersion: The purpose of the Container Closure Integrity test is to evaluate the barrier properties of filled vials and closed systems. The samples are tested by immersing media-filled containers into a microbial challenge for a specified amount of time using pressure and vacuum. The containers are then removed from the challenge, rinsed, incubated, and examined for growth. A bacteriostasis test is performed concurrent with the test or at the end of the seven-day incubation. Controls are performed with each challenge.

The selection of B. diminuta as the challenge organism is based on its very small size when grown under carefully controlled conditions. Due to its size, B. diminuta represents a most severe bacterial challenge. Other organisms may also be selected. The high concentration in microbiological challenge adds to the worst-case conditions for testing.

Samples must arrive sterile if filled by the sponsor.

B. diminuta is an aerobic organism so it is recommended, when filling test samples, to allow for an air headspace of ≥50% of the total volume of the container, if possible.

If you have additional questions about Dye and Bacterial Immersion testing services, or would like to consult with the experts at Nelson Labs, just send us a request or call us at +1 (801) 290-7500.