Bioburden and Endotoxin (LAL) Testing proposal help

[IHaveNoSpaceBar]

First and foremost, sorry for the long post.

I am part of a team involved with a new project at my company to manufacture a new product (basically IV-Bag). I am tasked with working on the Bioburden/Endotoxin (LAL) testing part for the product - as requested by customer.

The manufacturing steps include the manufacturing of the packaging material(film), converting it to bags(sealing/cutting), then installing the port piece needed to allow the end user to connect the tube to the bag. We will not be filling the bags with any solutions/liquids, another party will be doing this step.

I am at the beginning stage of the task, and really needed some guidance to help me figure out if I am on the right track or not. I ask that you please take a look at my proposal and point me in the right direction if you see any gaps. This type of project is a first for me and some/any assurance would be VERY appreciated.

Here is what I am planning to propose to my team, then the customer:

Based on lot sizes of Approx ~5000 parts, and ISO10993 sample requirements, I am proposing the following sample sizes to be tested at the beginning stages of the project:

- For the first month, I am proposing 10 samples from each lot to be tested for both Bioburden and Endotoxin(LAL). As far as I know, the 10 samples is based on the ANSI/ISO/FDA requirements.
a) Test 2 samples for lot sizes less than 1-30
b) Test 3 samples for lot sizes 31-100
c) Test 3% - not exceeding 10 samples for lot sizes equal or greater than 101

Since the lot size is greater than 101 (5000 total), I am going with (c) - 10 samples for each test.






- I am proposing 4 different kinds of Bioburden testing (don?t know if all are necessary-but according to my 3rd party lab, they are)
o Aerobic Bacterial
o Anaerobic Bacteria
o Spores
o Yeast & Mold

- I am proposing 1 LAL Endotoxin test - clotting type test - The 3rd party lab I deal with has mentioned that they are able to go down to 0.03 EU/ml sensitivity. I believe that this level is sufficient. I have noticed that the common standard is a maximum of 20 EU/ml per device. However, If I recall correctly, an IV-bag may fall in a more restricted category. Any input here would be appreciated.

- Initial testing cost estimates are very high. I have done some initial cost analysis to figure out how much the Bioburden and Endotoxin tests will cost per lot. Our 3rd party lab has mentioned $110 for the 4 test-bioburden analysis and about $110 for Endotoxin LAL test. If we are running production 8 times a month requiring 10 samples for bioburden and endotoxin (according to std) we are looking at a total of $17,600 per month. This is based on my calculation of (8 lots x 10 test samples x $220 bioburden & endotoxin cost) = $17,600.

- I am hoping that after the first month of production is over and hopefully all the results pass we can find an alternative to the expensive tests (I have already submitted some samples for initial baseline testing and the results are favorable)

- According to ANSI/AAMI ST 72 section 10 ?Bacterial endotoxin alternative to batch testing? there are alternatives to batch testing for product release through the demonstration that the manufacturing process and materials are well controlled. I am hoping that through the documentation of all the process steps and demonstrating that any bacteria cross contamination sources will be eliminated or drastically decreased. I am planning to capture and document this information in the form of an FMEA as well as cleaning, handling procedures.

- I would like to add that my opinion is that the manufacturing process of this product (IV bag) does not allow for bacteria cross contamination. A clean room environment will be used to assemble the port inside the sealed IVbag. The way I see it is that the port installation part of the process presents the biggest risk due to the frequency of occurrence(handling), risk, and detection (RPN). To control that risk, the operator will be wearing gloves, gowns, and following clean room procedures in general. I don?t know if a face mask is necessary for this operation. I just envision what if someone does sneeze near the product?

- Once a month?s worth of good test data comes back I plan on presenting it to the customer along with cleaning procedure, and FMEA so that we can throttle down our testing. I am hoping to throttle down the testing frequency to 10 samples for bioburden and 10 samples for endotoxin every 6 months, or possibly every quarter if 6 months is stretching it.



- In case the customer does not agree to my plan, I want to have a backup plan. I have seen that some companies choose to do Endotoxin testing in house. I have seen the name ?Pyrosate? a few times, which I believe is a machine manufactured by the association of cape cod, allowing inline endtoxin (LAL testing). The only issue is it not very sensitive, I have seen reports of the machine being able to do a minimum of 0.25 EU/ml. I have not seen in house bioburden testing options though.


As you can tell I am at the beginning stages of my plan and I am really hoping that some of the great minds on this board can help guide me through this if they see any gaps.

Much appreciated.

 

[Stijloor]

A Quick Bump!

Can someone help?

Thank you very much!!

 

[somashekar]

The manufacturing steps include the manufacturing of the packaging material(film), converting it to bags(sealing/cutting), then installing the port piece needed to allow the end user to connect the tube to the bag. We will not be filling the bags with any solutions/liquids, another party will be doing this step.

OK, is there an use of water in this process which may come in contact with the bag and tube (more so inside the bag and inside the tube)
What ISO class is your clean room maintained to ?

I have noticed that the common standard is a maximum of 20 EU/ml per device. However, If I recall correctly, an IV-bag may fall in a more restricted category. Any input here would be appreciated.

20EU/ml in device is for general application per USP, and is 10 times tighter ie 2EU/ml in device in case of use in the brain directly. See into the latest USP

- I am proposing 4 different kinds of Bioburden testing (don?t know if all are necessary-but according to my 3rd party lab, they are)
o Aerobic Bacterial
o Anaerobic Bacteria
o Spores
o Yeast & Mold

Depends upon your clean room classification and further the room people behavior. I would not do all these if you have established a good infrastructure. Surface swab test for gram -ve bacteria is what will interest me.
After all endotoxin is the outer layer of a dead gram -ve bacteria
The major route for the gram -ve bacteria is through water / moisture, and once these are killed in your sterilization process, the endotoxin remains there. So no water no trouble and sneeze, cough, bad practices in clean room are water contributors, which can kill your process. Take gaurd on this.

- In case the customer does not agree to my plan, I want to have a backup plan. I have seen that some companies choose to do Endotoxin testing in house. I have seen the name ?Pyrosate? a few times, which I believe is a machine manufactured by the association of cape cod, allowing inline endtoxin (LAL testing). The only issue is it not very sensitive, I have seen reports of the machine being able to do a minimum of 0.25 EU/ml. I have not seen in house bioburden testing options though.

We use pyrosate and is good for us. But we use it on a daily basis to test and pass the process water that is used in our device manufacturing. The co-relation of this result to the pooled endotoxin test results on finished device have been excellent and the end product results of pooled endotoxin tests has been 10 fold better. ie., while we have to meet <20EU/ml in the device, the end results on device have shown <2EU/ml consistently.

Good to have a laminar flow bench within the clean room and improve the class under the laminar hood where controlled work can be done for consistent good endotoxin results.

I do not know how you will conduct the endotoxin test on your device. The challenge here is extraction from device, and for this you will need endotoxin free water (pretty costly) and very controlled environment where subject experts can perform and give results acceptable to any 3rd party or customer.

(broken link removed) may be helpful to you ....

Also see the Similar Discussion Threads, down this page which can give you some more inputs. Hope all this helps you.

 

[petdressiva]

Hi All, I know this is a older thread, but I would like to ask if the ST72, USP 161, 85 or FDA Q and A dictate the frequency of LAL testing for medical device that is just in the warehouse. No movement or anything that can disturb the packaging and all. Can you guys advise me or reroute me to an existing guidance?

 

[planB]

Presuming you are talking about a terminally sterilized medical device stored in a warehouse, the quantity of bacterial endotoxins does not change over the validated shelf life. Thus, you would not re-test endotoxins through-out the shelf life because a shelf-life claims implies a sterile barrier system having been demonstrated to remain its integrity over this time period.

There might be different regulatory expectations for combination products that contain an active pharmaceutical ingredient: there, "pharmaceutical stability" validation activities may be expected to contain bacterial endotoxin testing at intermediate and final time points, even though these testing activities lack scientific justification for the reason given above.

HTH,

 

[petdressiva]

Thank you. Will you be able to share a study or guidance or any document for my reference to justify that endotoxin testing can be done once althoughout the device shelf life?

 

[planB]

AAMI ST72:2019, section 7.5 says the following:

7.5 For end-product testing, samples may be obtained prior to (pre-) sterilization or after (post-) sterilization. Post-sterilization samples encompass all of the factors that could affect the product or the endotoxin test.

If shelf life was a factor, it would be mentioned here.