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FOR EXPERT REVIEW PANEL USE ONLY
NOT FOR DISTRIBUTION
2
Method Safety Concerns:
Warning on handling diisopropyl ether should be added, since peroxide formation is a major concern as
an explosion hazard. (Not only that, but DIPE that has peroxides is not only dangerous, but will really
mess with the carotenoids). Assure it is stabilized and not stored so long the stabilizer protection has run
out.
Pros/Strengths:
Simple workup and isolation
Routine HPLC for a well equipped laboratory.
Cons/Weaknesses
Step E (b) . Concentration of beta carotene stock standard will be inaccurate. Simply measuring the
absorbance at 453 nm only tells you there is something absorbing at 453nm. HPLC must also be run
on the solution at 453 nm to measure all the peaks at 453nm, then correct the total absorbance for
the beta carotene absorbance by multiplying the total absorbance by the peak area of the beta
carotene over the total peak area at 453nm. Or by calculating the concentration based strictly on
absorbance and correcting by the ratio of the peak areas.
I assume working standard solution mentioned in G(b) is the HPLC standard solution prepared above.
Supporting Data
General Comment: Meets all the requirements of the SMPR (see the SMPR).
–
Method Optimization:
Performance Characteristics: Meets all the requirements of the SMPR (see the paper).
Analytical Range:
Carot-01 Review Forms