Leak Test for Capped Bottles/Vials

Dear all,
Does anyone give me any reference about leak test procedure for capped bottles or vials? We need the information about how many samples are needed during the capping process validation. Thanks in advance.


Requirement for parenteral product quality assurance, includes the demonstration of the microbiological integrity of container-closure system. This is frequently attempted by microbial challenge testing or the use of physical test methods such as bubble, dye ingress, pressure / vacuum decay, or electrical conductivity test.

Currently, there are no generally accepted methodologies that meet the requirement of demonstrating quantitatively, reliably and reproducibly that a parenteral product container-closure system is an effective barrier against microbial contamination, initially and throughout its shelf life.

We use online pressure regulated sealing system and also we perform the Microbial destructive test by picking 20 vials and perform in 2 lots.The vials are randomly picked.

Iam attaching 2 white papers on the testing method based on FDA gUIDANCE.


LIGHTHOUSEWhitepaperCCI.pdf (537.8 KB)

  1. ASTM D3078-02(2008), “Standard Test Method for Determination of Leaks in Flexible Packaging by Bubble Emission”; ASTM F1929- 98(2004), “Standard Test Method for Detecting Seal Leaks in Porous Medical Packaging by Dye Penetration; ASTM F2338-07, “Standard Test Method for Nondestructive Detection of Leaks in Packages by Vacuum Decay Method”; ASTM F88-07a, “Standard Test Method for Seal Strength of Flexible Barrier Materials”; ASTM F2097-05, “Standard Guide for Design and Evaluation of primary Packaging for Medical Products”; WK19936, “New Test Method for
    Indirect Measurement of Elastomeric Closure Compression Using an Automated Residual Seal Force Tester.”

  2. “Container and Closure Integrity Testing in Lieu of Sterility Testing as a Component of the Stability Protocol for Sterile Products,” FDA Guidelines, February 2008.

  3. ASTM F2391-05, “Standard Test Method for Measuring Package and Seal Integrity Using Helium as the Tracer Gas.”

Relevant Documents

[b][COLOR=“blue”]• ASTM D4991
• ASTM F2338-09

• U.S. Department of Health and Human Services, Food and Drug Administration, Center for Biologics Evaluation and Research, Center for Drug Evaluation and Research, Center for Devices and Radiological Health, Center for Veterinary Medicine. Container and Closure System Integrity Testing in Lieu of Sterility Testing as a Component of the Stability Protocol for Sterile Products, Guidance for Industry, 2008

• Ph. Eur.: Chapter 3.2.9; Rubber Closures for Containers for Aqueous Parenteral Preparations, for Powders and for Freeze-Dried Powders

• United States Pharmacopeia: <381> Elastomeric Closures for Injection
• United States Pharmacopeia: <1207> Sterile Product Packaging—Integrity Evaluation
• PDA Technical Report No. 27, (1998)[/color][/b]

Mr Prasad, thanks a lot for your information. I have read your process validation draft protocol for oral liquid dosage form and found the leak test at page 20. it is mentioned in the document that sampling for leak test are performed on the initial, middle, and end of the capping process. how much samples is appropriate for each time points?

[b]We sample every hour once.
8 to 10 bottles is the sample Size taken.

During the Final lot for such an inspection we pick up totally 20 bottles after Visual inspection.This process is just before labelling and arranging in individual cartons.Untill the test is cleared we do not give a releasing slip to packing.Out of 20 the failure must be just one bottle. If we have more than 1 failure, we immediately sample and another 20 and repeat the test. If it fails --then we will rethink what to do.

In a total batch of say 1500 liters each bottle size of 100 ml we will carry out 120 to 140cap integrity inprocess checking.[/b]

regarding the your draft protocol, there are 4 stages or sampling times: after machine setting, beginning of capping, middle of capping and at the end of capping. in this point we take 8-10 bottles. is it correct?
what are the meaning of ‘recapping’ and ‘seal breaking’?

What you mentioned is right regarding the sample collection.
Some times we find problems with capping-- They mightbe dented, they might be lose , they might be damaged etc etc–such places we break the seal and recap. This is just about 1% or less we get in a production batch. Such things will be recapped.

How do we fill the ‘recapping’ and ‘seal breaking’ columns of the table of data recording sheet V in your process validation protocol? Does ‘recapping’ mean every malformation or misplacement of the cap (rubber stopper) that need to be recapped with the new one, meanwhile ‘seal breaking’ mean malfunction of flip off vial/top cap/seal?

If there are some malfunctions in the equipment these things might happen.

They do not happen day in day out of production. Because machines are maintained and they will be kept in perfect condition to provide maximum output with minimum rejection of such capping or packing materials.

This is where Qualifying vendors come into picture.It makes sure that quality parameters of each supplied material meets the initial specifications that is mentioned in the quality agreement.

It does not fit into “V” diagram of process validation. You need to attach a new form by name “Risk based methods sheet”. In this all these thing will be entered properly.When ever you start up a batch – immediately after validation runs, you will run a batch size which is equall to production batch to show the shortest run time. In such an events you will note the pitfalls and lacunas of production and the production interruptions.Such interventions are noted in this document and they will also write CAPA. This will be available to topbrass of company management only.Most of parameters are given here.

I would like to understand that there is a lot of difference in quality, parameters and Regulatory restrictions between an Oral Bottle and a VIAL.

When ever you write a question please frame them properly and give the dosage form. I almost wasted some time about Vials.

Vials are different from bottles.

thanks a lot for the discussion. :slight_smile:

Dear all
This question was very helpful because i was also looking for the test vial number of CCI test.
Besides this question, i wonder the method validation of dye penetration test and microbial immersion test.
Acutually, i did not perform method validation because someone said that the controls functions corretly, it does not need method validation. But i am a little bit worried that this is not a perfect justification for not performing method validation.
Please provide me more information about method validation for CCI test??

Thanks in advance.


CCI test = Container Closure Integrity Test.
There are some people in FDA who looks very keenly into CCI testing.Some want Sterility test to be replaced by CCI testing. Looking If CCI test is done it is done just after product expires too.FDA looks into such data these days.Others ask CCI testing annually till the product expires.

Some manufacturers use the dye test to satisfy along with CCI data. Unfortunately, some FDA divisions still prefer the immersion test to the dye test.

FDA looks the data about method validation very keely on both Dye penetration test and Microbial ingress test for a new product launch or If the container closure system changes or if you change a manufacturing site.

CCI FDA Guideline attached

Attched guideline here

CCI FDA Guideline.pdf (64.2 KB)

Thank you very much for your answer.
Your answer is that we have to perform method validation for CCI test. Right??

If you available, can you provide the general method validation protocol for dye test and microbial test??

I am a little bit confusing the concepts of CCI method validation.

Thank you very much.

so, container and closure system integrity testing could be included in stability protocol as the replacement for sterility testing. thanks for the document, Mr Prasad.

Yes, Method validation should be carried out.
You must include the CCI in your stability program.
Guidelines are fast changing. You should mplement them as soon as you get an idea.

Thanq very much for the useful information and excellent discussion
Can you send me method development and validation protocol of CCI test for syringes and vials.

Dear all
i wanna to do a test about microbiology challenge for vial.
but i don’t know the method.choose which one Microorganism to soak.

thank you everyone.