| Radix Salvia Miltiorrhizae is the dried root of Salvia miltiorrhiza Bge.The main pharmacological active ingredients are the hydrosoluble components salvianolic acids and liposoluble components diterpenoid tanshinones.In this study,the correlativity of content from different sources,extraction technology,the main components and the stability of crude drug and extracts were investigated to reveal the interactions between the key components in different conditions.On the basis upon, formulation technology of bidirectional drug-loaded lipid microsphere of the two different polar components was studied.Also,the pharmacokinetics in rats after administration of single and mixed components in different ratios and components interactions were studied.According to Chinese Pharmacopoeia,cultivated and commericall available Salvia Miltiorrhizae from different habitats were identified and assayed.It was found that the source of Salvia Miltiorrhizae was single and there was no significant difference in thin-layer chromatography(TLC) as well as infrared spectrum.However, differences in contents of main index components were of significance.It was also found that the content of diterpenoid tanshinones existed a correlativity,as well as salvianolic acids.But there was no correlativity of content between tanshinones and salvianolic acids.Extraction technology of active ingredients with different polarity in Salvia Miltiorrhizae was investigated.The result showed that the active ingredients could be fully extracted after supercritical carbon dioxide extraction of diterpenoid tanshinones from Salvia Miltiorrhizae crude drug,then extracting alvianolic acids from the herb residue.The optimum condition of supercritical extraction was as follows:extracting pressure:25MPa,extracting temperature:50℃,extracting time:2.5h and ethanol was added as a cosolvent.The extraction process of salvianolic acids was discribed below. Soaking dry herb residue in 8 volumes of water for 4 hours,then decocting twice for 2 hours each time,95%ethanol was added to filtrate until the concentration of ethanol was 60%.After storage in cold for 40 hours,concentrated filtrate was eluted on a column of D101 macroporous resin by 50%ethanol.The eluate was finally freeze-dried.By studying the stability of Salvia Miltiorrhizae crude drug,liposoluble extracts and the standard methanol solution,it indicated that tanshinoneⅡA and cryptotanshinone were vulnerable to temperature,pH and light and prone to degrade. Compared with pure substances,other components in crude drug and extracts could delay the degradation of tanshinoneⅡA and cryptotanshinone.In addition, tanshinoneⅡA and cryptotanshinone were more stable under weak acid condition.According to the UPLC-ESI-MS chromatographic and mass spectral behaviors of Salvia Miltiorrhizae crude drug,salvianolic acids,diterpenoid tanshinones and reference substances,20 compounds have been separated and identified including 10 watersoluble components,7 liposoluble components,and 3 unknown components. Based on the research of ESI-MS fragmentation pathways of tanshinoneⅡA and salvianolic acid B at different collision energy,fragments of tanshinoneⅡA at m/z280, 277,262,252 and 249 in the positive ion mode were obtained.With respect to salvianolic acid B,fragments at m/z519,321,339 and 295 in the negative ion mode were formed.The in vitro degradation kinetics in rat plasma and liver homogenate of tanshinoneⅡA,cryptotanshinone in diterpenoid tanshinones,salvianolic acid B in salvianolic acids and mixed extract in the different proportion were investigated.The result indicated that with the increasing of plasma concentration,the degradation rate of 0.5mg/ml tanshinoneⅡA and cryptotanshinone in plasma was was decreased;the degradation rate of salvianolic acid B in 2.97mg/ml salvianolic acids in different concentration of plasma had no obvious change;the degradation rates of the three compounds in liver homogenate was high with the increasing concentration of liver homogenate.After mixing diterpenoid tanshinones and salvianolic acids in different ratio(20:1,10:1,5:1),the degradation rate of tanshinoneⅡA was obviously lowered. The degradation rate of cryptotanshinone and salvianolic acid B was slightly increased in 20%plasma and decreased significantly in 20%liver homogenate.In summary,interaction of drugs metabolism in plasma and liver homogenate occurred.Formulation,process and physicochemical properties of bidirectional Salvia Miltiorrhizae lipid microsphere injection were studied.It showed that the color of bidirectional drug-loaded lipid microsphere injection of Salvia Miltiorrhizae was dark brown.The particle size of the injectionwas about 200nm withζ-potential -20——40mv.The appearance,particle size andζ-potential had no significant change before and after sterilization.Salvianolic acids mainly distributed in water phase and diterpenoid tanshinones mainly distributed in oil phase.By intravenous and oral administration,rats pharmacokinetics of tanshinoneⅡA and salvianolic acid B from diterpenoid tanshinones,salvianolic acids and mixed extracts were studied.A UPLC/MS-MS methods of simultaneously determining tanshinoneⅡA and salvianolic acid B in plasma was established. Compared mixed extracted emulsions with tanshinones emulsions and salvianolic acids emulsions isodose,AUC of tanshinoneⅡA and salvianolic acid B was obviously increased by 2-14 times,where the AUC of low dosage group and middle dosage group increased to 2 times and the AUC of high dose group was increased to about 14 times and 5 times respectively(P<0.01).Furthermore,plasma clearance(CLt) decreased significantly(p<0.01) and the peak concentration(C0.083h) significant increased(p<0.01).Compared oral high dose and middle dose mixed extracts with tanshinones suspension and salvianolic acids suspension isodose,AUC of tanshinoneⅡA and salvianolic acid B significant increased(p<0.05),total plasma clearance(CLt) of tanshinoneⅡA and salvianolic acid B in high dose group and low dose group decreased significantly(p<0.01).In conclude,there existed interaction of drugs in metabolism in rats after mixing of the two extracts.
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