Let’s get the boring part out of the way first, then we can get to some chromatograms!
Cleaning validation is demonstrating with objective evidence that a cleaning process can repeatably and reliably remove all traces of product and any cleaning agents from equipment after the manufacturer of the batch.
The reason this is important is that if a cleaning method hasn’t been validated, then there can be traces of the last batch and any cleaning agents that were used in the next batch of material.
We recently came across an example of this, and after several weeks of testing, we arrived at a potential issue with cleaning validation. Let’s look at some chromatograms!
Here is the original chromatogram where we identified that the client’s BPC-157 capsules were low purity:
The peaks around the BPC-157 peak are typical, but the peaks at 1.5 minutes and 9 minutes were concerning. So we started testing the other components in the capsule mix to see if it was something in the sample matrix.
Nothing:
Still nothing:
Finally the empty capsules themselves, and we cannot find these peaks at 1.5 or 9 minutes anywhere:
So we put on our thinking caps, asked what other products the manufacturer produced and we ran those products on our BPC-157 test method.
Nope:
Nothing:
Bingo! Anastrozole capsules have the same peaks we saw in the BPC-157 capsules at 1.5 minutes and 9 minutes on our BPC-157 method:
While we cannot confirm that either of these peaks are anastrozole without mass spec or an anastrozole reference standard, we can confirm that there is something in the anastrozole capsule mix that is not being cleaned off the machine between runs.
If it is anastrozole, then users taking the BPC-157 capsule product are unknowingly taking a breast cancer / estrogen reduction treatment compound as well.
The Takeaway
Effective machine cleaning between runs is critically important. Understanding the solubility of the materials in the formula and making sure cleaning procedures address those compounds is a great approach to ensuring effective cleaning.
Anastrozole is only slighly soluble in water, so if this manufacturer is using water to clean their equipment, it won’t be as effective as using something anastrozole is freely soluble in like methanol, ethanol, or acetone.
When you are investigating a new manufacturer, talk to them about their machine cleaning procedures. Ask them if they have any cleaning validation or cleaning testing.