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- Posts: 1
- Joined: Thu Aug 02, 2012 5:50 am
I'm trying to get linearity in plasma standard of the compound with 3-aminopyrazole moiety.
All compounds having 3-aminopyrazole showed exactly same pattern in the standard as smaller peak area, lower standard concentration resulting in standard failure.
Also, peak was separated to two leaks (first eluted peak area was bigger at lower concentration and later eluted peak area was bigger at higher concentration).
Column is Phenomenex Hydro RP C18 and Waters Acquity UPLC BEH C18. Mobile phase I'm using is 0.1% FA in water/0.1% FA in ACN. Plasma samples were acidified with FA to make one peak (I think this is due to tautomerism), but it didn't help at all. Do you have any idea to resolve this issue?
Many thanks in advance,
All compounds having 3-aminopyrazole showed exactly same pattern in the standard as smaller peak area, lower standard concentration resulting in standard failure.
Also, peak was separated to two leaks (first eluted peak area was bigger at lower concentration and later eluted peak area was bigger at higher concentration).
Column is Phenomenex Hydro RP C18 and Waters Acquity UPLC BEH C18. Mobile phase I'm using is 0.1% FA in water/0.1% FA in ACN. Plasma samples were acidified with FA to make one peak (I think this is due to tautomerism), but it didn't help at all. Do you have any idea to resolve this issue?
Many thanks in advance,
