Isolation and quantitative analysis of cryptotanshinone, an active quinoid diterpene formed in callus of Salvia miltiorrhiza BUNGE.
Effect of N(6)-benzyladenine (BA) on tanshinone formation in callus cultures of Salvia miltiorrhiza was examined in an attempt to increase the productivity of the medicinal compound, cryptotanshinone. Primary callus was induced by culturing leaf explants on Murashige and Skoog's (MS) basal medium supplemented with 1.0 mg l(-1) of 2,4-dichlorophenoxyacetic acid (2,4-D) in darkness. The callus proliferated further on MS basal medium containing 1.0 mg l(-1) 2,4-D and 0.5 mg l(-1) BA and was analyzed for cryptotanshinone by high performance liquid chromatography (HPLC). The HPLC results indicated that it contained small amounts of cryptotanshinone (0.26+/-0.05 mg/g dry wt). Omission of 2,4-D from the medium resulted in a marked increase in the content of cryptotanshinone in callus. The HPLC analysis revealed that the content of cryptotanshinone in the callus cultured on the MS basal medium supplemented with 0.1, 0.2, 0.5, 1.0, and 2.0 mg l(-1) of BA was significantly higher than the marketed crude drug (processed underground parts of S. miltiorrhiza). Maximum yield of cryptotanshinone (4.59+/-0.09 mg/g dry wt) was observed in the callus cultured on MS basal medium supplemented with 0.2 mg l(-1) BA for 60 d. Cryptotanshinone was isolated from callus through silica gel column chromatography followed by preparative TLC and characterized based on NMR and mass spectral data.
Wu
Chin-Tung
CT
Department of Agronomy, National Chung Hsing University, Taichung 402, Taiwan.
Mulabagal
Vanisree
V
Nalawade
Satish Manohar
SM
Chen
Chung-Li
CL
Yang
Te-Fang
TF
Tsay
Hsin-Sheng
HS
D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology; D001593:Benzyl Compounds; D002851:Chromatography, High Pressure Liquid; D004224:Diterpenes / Q000302:isolation & purification*; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000302:isolation & purification*; D007701:Kinetin; D010616:Phenanthrenes / Q000302:isolation & purification*; D010936:Plant Extracts; D010937:Plant Growth Regulators / Q000494:pharmacology; D018515:Plant Leaves / Q000737:chemistry / Q000254:growth & development; D010946:Plants, Medicinal / Q000737:chemistry* / Q000254:growth & development; D011687:Purines; D027561:Salvia miltiorrhiza / Q000737:chemistry* / Q000254:growth & development
D001593:Benzyl Compounds; D004224:Diterpenes; D004365:Drugs, Chinese Herbal; D010616:Phenanthrenes; D010936:Plant Extracts; D010937:Plant Growth Regulators; D011687:Purines; C037886:cryptotanshinone; D000225:Adenine; C480551:benzylaminopurine; D007701:Kinetin