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- Posts: 58
- Joined: Tue Oct 09, 2012 12:25 am
Does anyone know why the USP calls for the resolution solution (pre-vitamin D3) to be made? I thought it was to ensure the HPLC would detect the thermal degradation of D3. We put both the working standard and the test samples in a waterbath at 60 deg C for 1hour. If both the pre-vitamin D3 and D3 can be utilized by the body and convert into the active 1,25-dihydroxycholecalciferol, then would this be reported as total Vitamin D3?
I was able to see the pre-vitamin D3 separation from the D3 using the USP method 1 for Vitamin D3 in oil capsules. I am working on creating a UPLC/MS method for D3. I have my D3 peak coming out at 2.5 minutes. Waters has a method for creating derivatives in order to quantitate the pre-vitamin D3 and D3. Various other labs that I checked with are unable to separate these isomers. Any feedback would be great.