The Salvia divinorum Research and Information Center - Shroomery

PHYTOCHEMICAL ANALYSIS
quantify precisely the amount of salvinorin A present in samples of S. divinorum
or any other plant species. The purpose of the current study was to develop a
sensitive quantitative assay to determine the amount of 1 present in leaf samples of
S. divinorum. In addition, the method has been used to screen for this compound in
other species.
EXPERIMENTAL
General experimental procedures. Salvinorin A (1) used for standard curve
development was purified from S. divinorum leaf extracts and authenticated by
nuclear magnetic resonance spectroscopy and by thin layer chromatographic
(TLC) comparison with authentic 1 (Valdés et al., 1994). All solvents used for
extraction and chromatography were of high performance liquid chromatographic
(HPLC) grade from Fisher Scientific (Fair Lawn, NJ, USA). Water used in HPLC
mobile phase mixtures was distilled and subsequently filtered through a 0.22 ~im
membrane (Millipore Corp., Bedford, MA, USA).
Plant materials. Asexually propagated plants of S. divinorum used in establishing
the HPLC methods were purchased from Logee's Greenhouses (Danielson, CT,
USA) and multiplied by rooting additional cuttings in the S.B. Penick
Experimental Greenhouse at the Philadelphia College of Pharmacy and Science.
Voucher specimens were deposited in the herbarium of the Philadelphia College of
Pharmacy and Science. Additional S. divinorum samples used for analysis were
collected from established outdoor stands.
Extraction of samples. Plant tissues were lyophilized after harvest and ground to
a homogeneous powder in a Wiley mill (no. 20 mesh). Samples (0.500 g) of
lyophilized whole leaf were extracted in 125 mL roundbottom flasks by steeping in
25 mL of chloroform for 30 min. The extract was filtered and the filtrate set aside.
The extraction flask and filtered solids were rinsed with an additional 15 mL of
fresh chloroform. The filtrate from the rinse was then combined with the original
filtrate and the resulting solution was evaporated to dryness with a rotary
evaporator. The dry solids were redissolved in a mixture of 20.0 mL methanol and
5.0 mL acetone using sonication to assist in dissolving of all solid material.
Chromatography conditions. HPLC was performed using a Milton Roy HPLC
system (Riviera, FL, USA), consisting of a Constametric 3000 Series isocratic
pump, a Rheodyne injector (Rheodyne L.P., Cotati, CA, USA), and a
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