Isolation and Quantitative Analysis of Cryptotanshinone, an Active Quinoid Diterpene Formed in Callus of <i>Salvia miltiorrhiza</i> B<small>UNGE</small>

Abstract

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.

Activation tagging in Salvia miltiorrhiza can cause increased leaf size and accumulation of tanshinone I and IIA in its roots

“…In previous studies, in vitro methods such as hairy root cultures (Hu et al 1993; Ge and Wu 2005a; Yan et al 2005), cell suspension cultures (Chen et al 1997), addition of elicitors and optimization of media (Wu et al 2003; Zhang et al 2004; Ge and Wu 2005b; Gupta et al 2011), have been reported. In our previous study, ATM transgenic callus lines of Salvia with greater tanshinones content were developed (Lee et al 2008).…”

“…miltiorrhiza has been studied by various research groups over the world, especially in biotechnological and in vitro propagation field. Through several strategies like genetic manipulation of biosynthetic pathway via genetic transformation (Chen et al 1997; Lee et al 2008; Kai et al 2011), hairy root cultures (Hu et al 1993), use of plant growth regulators and elicitors (Wu et al 2003; Zhang et al, 2004; Ge and Wu 20052005; Gupta et al 2011), success has been reported in enhancing the production of secondary metabolites in S. miltiorrhiza .…”

Rapid separation and identification of phenolic and diterpenoid constituents from Radix Salvia miltiorrhizae by high‐performance liquid chromatography diode‐array detection, electrospray ionization time‐of‐flight mass spectrometry and electrospray ionization quadrupole ion trap mass spectrometry

“…The dried root of Dan‐shen (Radix Salvia miltiorrhizae ) is one of the most popular traditional Chinese herbal medicines and has been widely used for the treatment of menstrual disorders, menorrhealgia, insomnia, menostasis, blood circulation diseases, and other cardiovascular diseases 1–9. Many studies of its chemical constituents have been reported 10–16. Depending on their chemical structures, the major bioactive constituents can be classified into two major groups: water‐soluble phenolic compounds and nonpolar diterpenoid compounds.…”

A Mechanistic Study of the Intestinal Absorption of Cryptotanshinone, the Major Active Constituent ofSalvia miltiorrhiza

“…CTS (purity Ͼ99%, as determined by thin-layer chromatography) and tanshinone IIA (purity Ͼ95%, as determined by thin-layer chromatography) were extracted and purified from the root of S. miltiorrhiza using the CH 2 Cl 2 method as described previously (Hur et al, 2005) at the School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, People's Republic of China. CTS and tanshinone IIA were identified by comparison of liquid chromatography-mass spectrometry and nuclear magnetic resonance spectral data with reference values (Wu et al, 2003). Phenol red was purchased from Tianjin Chemical Reagents Institute (Tianjin, People's Republic of China).…”