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Primary and Secondary Evaluation of Method SJW-03
SJW-03:
Determination of St. John’s wort Components in Dietary Supplements and Functional
Foods by Liquid Chromatography
Author(s):
Catharina Y.W. Ang, Yanyan Cui, Hebron C. Chang, WenhongLuo, Thomas M. Heinze,
Lawrence J. Lin, and Antonia Mattia, U.S. Food and Drug Administration, National Center
for Toxicological Research, Division of Chemistry, HFT-230, 3900, NCTR Rd, Jefferson, AR
72079,, U.S. Food and Drug Administration, Center for Food Safety and Applied
Nutrition, Office of Food Additive Safety, Division of Petition Review, HFS-265, 5100 Paint
Branch Pkwy, College Park, MD 20740
S
UMMARY OF
M
ETHOD
:
G
ENERAL
C
OMMENTS
:
The method presents results dealing with the extraction and the determination of four (4) bioactive St.
John’s wort compounds (pseudohypericin (PHP), hypericin (HP), hyperforin (HF), and adhyperforin
(AHF)) in SJW aerial parts (leaves and flowers), dietary supplements and functional foods such as SJW
capsules, tea bag, snack bar, puff and drinks. The compounds were well separated through the
developed HPLC method using an isocratic elution program. The starting material was sonicated in a
methanolic solution and the obtained extract was analyzed through LC/UV at 2 wavelengths (290 & 590
nm) and LC/MS analysis. Two standard calibration curves with high and lowlevel ranges were
constructed for the quantitative determination of target compounds using BKF as an internal standard.
P
ROS
/S
TRENGTHS
:
This method has good separation and detection techniques such as HPLC, UV, and MS allowing the
separation and quantitative analysis of individual SJW bioactive compounds. This method could be
applied to several matrixes.
C
ONS
/W
EAKNESSES
:
No internal standard was used for hypericin, pseudohypericin, and some samples (Snack bar).
Confirmation by LC/ESI/MS should be performed using the same chromatographic conditions. The
retention time changes due to the column and room temperature variation and also to the used mobile
phase pH variation. Recovery test should be performed on blank matrixes.
EXPERT REVIEW PANEL VOTE
AND
RECOMMENDATION
MOTION: Not to make a decision on this method regarding First Action Official Method status
pending review of results validation data for method suitability for hyperforin determination.
(Brown, Sharaf – Unanimous)
Amended: This method is not recommended for First Action official Method status based on the data
presented. (Brown, Sharaf – Unanimous) Motion Passed
ERP PROFILE SUMMARIES
85