Regarding tests results, we would like to comment, that it seems that
test method wasn't incorrect: "I would have done 50:50 ACN:H20, but
the ACN was almost out".
According to your test method, concentration of ACN is too high,
sample can't be eluted, so that's why the ACN was almost out!
The appropriate testing environment should be as follows:
Column : Xbridge BEH130 C18,4.6*250mm,5um
Solvent A : 0.1% Trifluoroacetic in 100% Acetonitrile
Solvent B : 0.1% Trifluoroacetic in 100% Water
Gradient : A B
0.0min 22% 78%
25.0min 47% 53%
25.1min 100% 0%
30.0min Stop
Flow rate : 1.0ml/min
Wavelength : 220nm
We confirm that our products are at least 98% pure and each batch is
tested with HPLC after production cycle.
Hopefully our explanation will solve questions.