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- Joined: Tue Mar 10, 2020 1:03 pm
I have developed an HPLC method for testing Benzalkonium chloride in a finished product formulation of a hand soap. The HPLC method uses the following setup:
Column: Zorbax Bonus-RP from Agilent
Eluent A: Acetonitrile + small amount of TFA
Eluent B: Water + small amount of TFA
UV detector @ 263nm
The method uses the peak area from the C-12 formula along to determine the concentration in the finished product. The calibration of the system is done using the specific lot of Benzalkonium chloride that the product was batched with so that way we are normalizing the result based on the C-12 concentration in the raw material.
The method passed for specificity, linearity & limit of detection. The method was validated with a high level of accuracy and precision. The method is used to analyze finished products developed with a concentration of ~0.12% BZK.
The reason we did not integrate the C-12, C-14 and C-16 homologues is because specific finished product formulations exhibit coelution at the retention times for the larger homologues.
We have successfully tested finished product formulations for awhile using the method without any issues. However, recently we tested some aged materials and noticed a consistent increase in the assayed value from their original result. Across 1 to 2 year samples we saw an increase in 0.003% to 0.010%. BZK. After further investigation, we tested the placebo/specificity sample and saw a small peak at the C-12 retention time.
This was interesting as the original testing of that exact sample showed no peak at the C-12 retention window. We have repeated this observation for a number of placebo samples at different time points. I believe the earliest we saw the peak show up was around the 6-8 month time point.
Does anyone have any idea as to why we are experiencing this phenomenon?
Is it typical to test aged samples during a method validation process?
Column: Zorbax Bonus-RP from Agilent
Eluent A: Acetonitrile + small amount of TFA
Eluent B: Water + small amount of TFA
UV detector @ 263nm
The method uses the peak area from the C-12 formula along to determine the concentration in the finished product. The calibration of the system is done using the specific lot of Benzalkonium chloride that the product was batched with so that way we are normalizing the result based on the C-12 concentration in the raw material.
The method passed for specificity, linearity & limit of detection. The method was validated with a high level of accuracy and precision. The method is used to analyze finished products developed with a concentration of ~0.12% BZK.
The reason we did not integrate the C-12, C-14 and C-16 homologues is because specific finished product formulations exhibit coelution at the retention times for the larger homologues.
We have successfully tested finished product formulations for awhile using the method without any issues. However, recently we tested some aged materials and noticed a consistent increase in the assayed value from their original result. Across 1 to 2 year samples we saw an increase in 0.003% to 0.010%. BZK. After further investigation, we tested the placebo/specificity sample and saw a small peak at the C-12 retention time.
This was interesting as the original testing of that exact sample showed no peak at the C-12 retention window. We have repeated this observation for a number of placebo samples at different time points. I believe the earliest we saw the peak show up was around the 6-8 month time point.
Does anyone have any idea as to why we are experiencing this phenomenon?
Is it typical to test aged samples during a method validation process?
