Candidates for 2016 ERP of the Year

ERP PROFILE SUMMARIES

Primary and Secondary Evaluation of Method SJW-02

SJW-02: Evaluation of major active components in St. John’s wort dietary supplements by high- performance liquid chromatography with photodiode array detection and electrospray mass spectrometric confirmation Frances F. Liu\ Catharina Y.W. Ang*, Thomas M. Heinze, Joshua D. Rankin2, Richard D. Beger, James P. Freeman, Jackson O. Lay Jr., US Food and Drug Administration, National Center for Toxicological Research, Division of Chemistry, 3900 NCTR Road, Jefferson, AR 72079, USA This method was published in 2000 and used RP-HPLC/DAD for the quantification of the flavonoids (rutin, hyperoside, isoquercitrin, quercitrin, and quercetin), phloroglucinol (hyperforin), and naphthodianthrones (hypericin and pseudohypericin) in five capsule products of St. John's wort. LC-ESI MS was used for confirmation of the detected compounds. Samples are extracted with an ethanol- acetone solution via shaking in a water bath at 55°C for 5.6 hours. Extract is filtered through filter paper into volumetric flask and diluted to 50.00 mL prior to analysis by reversed phase HPLC. Chromatographic separation is achieved on a 3 μm 250 x 4 mm C18 column at 25°C employing a gradient with 0.5%TFA/water and 0.5% TFA in methanol/acetonitrile (13:7). Analytes are detected by photodiode array at 284 and 590 nm and quantified using luteolin as an internal standard with peak confirmation by subsequent electrospray ionization mass spectrometry. The LC-MS method is 125 minutes long, employs a different chromatographic conditions, as opposed to detection in series, and as such offers no real advantage. P ROS /S TRENGTHS : This method used LC-ESI MS to confirm the identity of the peaks and the purity of the standards was confirmed by UV, MS and NMR. It detects and quantifies the analytes of interest and has a simple extraction procedure. C ONS /W EAKNESSES : This method has a long extraction procedure and HPLC run time. Only capsules were used in this method and the composition of those capsules was not detailed. There were challenges regarding the precision and stability. The LOQ, RSD r , RSD R did not appear to meet the SMPR. There was no recovery or LOD data provided. Author(s): S UMMARY OF M ETHOD : G ENERAL C OMMENTS :

EXPERT REVIEW PANEL VOTE AND RECOMMENDATION

MOTION: Not to consider this method for First Action Official Method status. Brown, Schaneberg (Unanimous) Motion Passed

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