Ultrasound-assisted Deep Eutectic Solvents Extraction Of Tanshinone ⅡA From Salvia Miltiorrhiza And Its Anti-LDL Oxidation Modification

Salvia miltiorrhiza belongs to the genus Salvia miltiorrhiza of Labiatae,which is an important Chinese medicine for the treatment of cardiovascular diseases.Due to its pharmacological effects such as anti-oxidation,anti-coagulation,anti-atherosclerosis and anti-apoptosis,tanshinoneⅡA is the main effective component.Deep eutectic solvents have the advantages of simple preparation process,low cost,biodegradable,and 100%atomic utilization rate in the synthesis process.They conform to the concept of green chemistry and are considered as good substitutes for traditional volatile organic solvents and ionic liquids.In this study,the green and efficient extraction and purification technique for determination of tanshinoneⅡA from Salvia miltiorrhiza with a combination of ultrasound-assisted hydrophobic DESs extraction（UAE-DESs）and macroporous resin enrichment were developed.On this basis,the anti-low density lipoprotein（LDL）oxidation activity of Salvia miltiorrhiza extracts was evaluated by in vitro Cu²⁺induction on LDL.The main results are as follows:1.Thirteen hydrophobic DESs were successfully prepared by using methyl trioctyl ammonium chloride as hydrogen bond acceptor,five organic acids and eight organic alcohols as hydrogen bond donors.Combined with ultrasonic assisted extraction（UAE）technology,the effects of the composition of DESs and its molar ratio on the extraction yield of tanshinoneⅡA in Salvia miltiorrhiza Bunge were investigated.The results indicated that the hydrophobic DESs composed of methyl trioctyl ammonium chloride and n-propanol（molar ratio of 1:4）showed higher extraction efficiency for tanshinoneⅡA in Salvia miltiorrhiza.2.The UAE process of tanshinoneⅡA in was optimized by the single factor experiment and Box-Behnken design（BBD）.The maximum extraction yield of1.109±0.007 mg/g was obtained at a extraction temperature of 44°C,an extraction time of47 min and a liquid-to-solid ratio of 40 mL/g.Compared with common methods,it has the advantages of less solvent consumption,high extraction efficiency and environmental protection.3.The adsorption and elution properties of seven macroporous adsorbents,X-5,AB-8,D101,D4020,DM130,HPD100 and HPD400A,for tanshinoneⅡA in DESs extracts were investigated.It was found that HPD400A had the best enrichment and purification effects.Under the optimal enrichment and purification conditions,the adsorption rate of tanshinoneⅡA in DESs was 96.45%,and the elution rate was 80.45%,which had a higher recovery rate.The recovery rate of the eutectic solvent was 96.83%.It indicates that tanshinoneⅡA could be effectively recovered and enriched from DES extracts.After recovered,the hydrophobic eutectic solvent has good integrity and can be reused.4.The property of anti-LDL oxidation of tanshinoneⅡA crude extracts was studied by determining the effects of conjugated dienes,TBARS and lipofuscin.in vitro Cu²⁺induction on LDL.The results showed that with the increase of the concentration of the Salvia miltiorrhiza extracts,the formation of conjugated diene,TBARS and lipofuscin was significantly inhibited,and the delay time of LDL oxidation was prolonged in a dose-dependent manner.The results showed that Salvia miltiorrhiza could effectively alleviate the oxidative susceptibility of LDL,reduce the degree of lipid peroxidation of LDL,and have a certain anti-LDL oxidation effect.