A-03
Modification of a method for isolation of teucrin A and other diterpenoids from authenticated Teucrium Chamaedrys L.
P. R. Sundaresan1 , S. Slavoff2 , E. Grundel1 , K. White1 , J. I. Rader1 , 1FDA, 2JIFSAN
Neoclerodane diterpenoids are present in Teucrium species, commonly known as germander. Germander has been used in folk medicine as a stimulant, diaphoretic and a diuretic. It has been marketed in European countries as an adjuvant to weight control. Severe hepatotoxicity resulted from its use and preparations containing germander have been prohibited in France since 1992. Previous reports from this laboratory have shown that teucrin A and other diterpenoids (e.g. teucrin 5) can be isolated from the aerial parts of authenticated T. chamaedrys L. by HPLC and identified by LC/MS. For the purpose of isolation and characterization of teucrium diterpenoids on a preparative scale, we adopted a solid phase extraction (SPE) method for concentrating large volumes of teucrin A and teucrin 5 fractions obtained after semi-preparative HPLC of methanol extracts. For SPE of teucrin A, we employed a 500 mg styrene divinylbenzene tube (Phenomenex, Torrence, CA). The average recovery of teucrin A standard was 100.9 % in two separate experiments. For SPE of teucrin 5, we used Strata -X sorbent tube and the average recovery was 101.2%. The semi-preparative HPLC of the methanol extract of aerial parts of teucrium plant powder was carried out using a 250x 10 mm Synergi 4 Max-RP 80 A column (Phenomenex, Torrence, CA). The column was monitored at 220 nm with a diode array detector and fractions were collected with a Beckman SC-100 fraction collector. These procedures improved the efficiency of the isolation of in-house standards for diterpenoids.
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Last updated on 2004-APR-02 by frf