Benzalkonium chloride (BKC) is Quaternary ammonium compound(disinfectant) and cationic surafctant...used in pharmaceutical industry so we want to test Bacterial endotoxin in (BKC). We did LAL method(limit:2.5 EU/ml, Lysate sensitivity 0.03) but didnt get positive control(No gel formation) and also check by Kinetic chromogenic method but not get the results as BKC have strong disinfectant property.Tween 80 and Lecithin neutralizes this activity so can we use that in LAL method( In dilutions or wherever applicable) and how?
You should not use Tween 80; All consumables used for LAL test must be endotoxin free; BAC has inhibiting nature, we can rectify this problem with more dilution with LRW, Could you explain more details about your test product (conc. & use)?
Dilution generally we did in LAL assay are as below:
MVD = Endotoxin limit × Concentration of sample solution / λ
Where λ = Sensitivity of Lysate (0.03)
= 2.5 EU/ml x 1ml/ml / 0.03 = 1: 80
MVD/2 = 1: 40
Sample preparation:
MVD/2 dilution: 0.1 ml Sample + 3.9 ml LRW ,also we did analysis at MVD/4.
Dilute the sample and shake on vortex shaker for 3 minutes.
Concentration of BKC that we have is 50% and it is mainly used in pharmaindustry as a preservative.
Hello all,
After lots of experiments done on Benzalkonium chloride to perform BET, here is the solution which we found out to perform BET.
As any pharmacopoeia didnt provide limit of BET testing, we developed in house limit(0.5 EU/mg) to perform this testing and we have got excellent results without adding tween 80 or lecithin.
We used Kinetic chromogenic method-PTS instrument from Charles river company. It has lysate sensitivity in their cartridge about 0.5- 0.005. It is totally reliable method to perform BET and gives result in just 15 min. you can search for Endosafe nexgen PTS user guide for complete information.
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