Abstract
Dihydrotanshinone, tanshinone I, cryptotanshinone, and tanshinone IIA are major lipid-soluble constituents isolated from Salvia miltiorrhiza Bunge (Danshen). In the present study, a systematic method was developed to simultaneously isolate and purify those compounds using macroporous adsorption resins and semi-preparative HPLC with a dynamic axial compress (DAC) system. The Danshen extract (95% alcohol) was divided into three fractions using different concentrations of alcohol (0%, 45%, and 90%) on D101 column. The content of total tanshinones of 90% alcohol eluent (TTS) was over 97%. Furthermore, the anti-inflammatory effects of those samples were investigated on LPS-stimulated RAW264.7 cells and three animal models. The results showed that the anti-inflammatory effect of TTS in vitro was superior to the one of any other sample including 0% and 45% eluent, and total tanshinones capsules. In addition, TTS exhibited a stronger anti-inflammatory effect than that of dihydrotanshinone, tanshinone IIA, cryptotanshinone, and tanshinone I, respectively. For animal models, TTS could significantly suppress xylene-induced ear oedema and rescue LPS-induced septic death and acute kidney injury in mice. In summary, the separation process developed in the study was high-efficiency, economic, and low-contamination, which was fit to industrial producing. TTS is a potential agent for the treatment of inflammatory diseases.
Highlights
The inflammation is always triggered by damage to organisms, which plays a defensive role in injury or infection[1]
In China, total tanshinones capsules (TTC) that were just prepared by 95% alcohol extract were employed as an anti-inflammatory medicine in the market (Z13020110)
An effective preparative method was developed to simultaneously isolate and purify DTAN, tanshinone I (TANI), CTAN, and tanshinone IIA (TANA) (Fig. 1) using macroporous adsorption resins (MARs) and preparative reversed-phase HPLC with a dynamic axial compression (DAC) column system, which provides a novel method for large-scale purification of tanshinones from natural resources
Summary
The adsorption capabilities of MARs are associated with the polarity or chemical structure of adsorbates, and with the characteristics of adsorbents such as the unique surface area, pore diameter, pore volume and so on, and even with adsorption medium[13]. Non-polar resins are more applicable to adsorption of them. Seven MARs (Table 1) were employed to determine the static adsorption and desorption capability. In terms of our experiments, the non-polar resin D101 and HPD100 exhibited higher adsorption than other resins (Table 2), indicating that the similar polarity with tanshinones and the smaller pore size of resins for adsorption were optimal.
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