Analysis Of Chemical Constituents Of Shenmai Injection

Shenmai injection is a sterile water solution which is composed of red ginseng extracts and Ophiopogon japonicus extracts.It has the benefits of making our body stronger and nourishing our body.It is clinically used in the treatment of shock caused by weak,coronary heart disease,viral myocarditis and so on.For traditional Chinese medicine injections,due to factors such as different origins of prescription medicinal materials,processing techniques of medicinal materials and differences in preparation processes,it often causes differences on the chemical components and their contents in injections,which may affect the efficacy and even produce toxic side effects.In order to ensure the controllable,safe and effective quality of traditional Chinese medicine injections,it is firstly necessary to clarify the chemical components in the injection as much as possible,determine the content of the injections,establish corresponding fingerprints,and find an effective method to control the quality stability of traditional Chinese medicine injections from the source.Ensuring that Chinese medicine injections are safer and more effective,this paper qualitatively and quantitatively analyzed the chemical constituents of ginsenosides and Ophiopogon japonicus saponins in Shenmai injection,and establishes new analytical methods based on different sample pretreatment techniques.Firstly,qualitative analysis of rare ginsenosides in Shenmai injection is carried out,and seven kinds of rare ginsenosides are analyzed by solid phase extraction?SPE?-high performance liquid chromatography?HPLC?,which are 20?S?-Rh₁,Rg₆,Rk₃,20?S?-Rg₃,20?R?-Rg₃,Rk₁,Rg₅.The experimental conditions are optimized to establish a method of a solid phase filled with 1.8g ODS,6mL50%aqueous methanol solution for eluent and 6 mL methanol solutionfor eluent.The detection limits of 7kinds of rare ginsenosides are in the range of 0.07¹.47?g/mL,and the recoveries are in the range of 96.2%¹05.1%?n=6?,and the RSDs range are 1.3³.1%?n=6?.The experimental results showed that the contents of above seven kinds of rare ginsenosides in the Shenmai injections made by different manufacturers are given as follow:20?S?-Rh₁:13.45¹5.95?g/mL;Rg₆:7.62¹0.75?g/mL;Rk₃:2.35⁵.04?g/mL;20?S?-Rg₃:11.20²9.70?g/mL;20?R?-Rg₃:8.34¹9.29?g/mL;Rk₁:16.75²2.90?g/mL;Rg₅:65.25¹58.99?g/mL.The results show that the highest content is ginsenoside Rg₅,the lowest content is ginsenoside Rk₃,but the contents of 7 kinds of rare ginsenosides in different manufacturers'Shenmai injection are different,especially the difference of ginsenoside Rg₅ is the biggest.It may be due to different sources of raw materials and differences in preparation processes.Secondly,this paper qualitatively analyzed the rare ginsenosides in red ginseng.Seven kinds of rare ginsenosidesare detected in red ginseng which had been detected in Shenmai injection,indicating that the above seven rare ginsenosides in Shenmai injectionare come from red ginseng.Then,in order to clarify whether the above seven kinds of rare ginsenosides in red ginseng are derived from the processing of red ginseng,the above seven kinds of rare ginsenosides in ginseng are qualitatively analyzed and four kinds of rare ginsenosides are detected.For the purpose of preventing the ginseng ginsenosides in ginseng change into rare ginsenosides under high temperature and enhancing Concentration Factor,a method of deep eutectic solvent aqueous two-phase extraction assisted by ultrasonic combined HPLC was established.After inspected the influence factors,the optmal conditions was showed as follow:at room temperature,1 mL deep eutectic solvent,2 mL deionized water,3.5g K₂HPO₄,ultrasonic treatment for 25 min.The experimental results showed that there are four kinds of rare ginsenosides in ginseng,which are 20?S?-Rh₁,Rg₆,Rk₃,20?S?-Rg₃.The detection limits of this method are 0.09⁰.30?g/mL,and the recoveries are in the range of 100.3%¹27.6%?n=6?,and the RSDs range are from 3.6%to 6.6%?n=6?.The results showed that the contents of 20?S?-Rh₁,Rg₆,Rk₃,20?S?-Rg₃ in ginseng are 74.27?g/g,2.58?g/g,2.71?g/g,0.34?g/g.The contents of the four rare ginsenosides in red ginseng are80.59?g/g,4.84?g/g,9.80?g/g,9.21?g/g.Compared with ginseng,there are more than three kinds of rare ginsenoside in red ginseng,and the four rare ginsenosides are also higher than that in ginseng.It indicated that in the process of preparing red ginseng by ginseng,some common ginsenosides are transformed to rare ginsenosides at high temperature.Finally,in order to explore the chemical constituents of Ophiopogon japonicus in Shenmai injection,the method of vacuum freeze-drying combined with HPLC-ELSD is used to qualitatively analyze the saponins and flavonoids in the injection.The contents of 2 kinds of Ophiopogon saponins?DT-13 and Ophiopogon saponin D?in the injection were quantitatively analyzed.It is carried out by the method of pre-freezing at-30°C for 5 min,-43°C and 0.09 mbar under vacuum freeze-drying for 2h.The detection limits of two kinds of Ophiopogon saponins in this method are in the range of 1.63³.45?g/mL,and the recoveries are in the range of 96.6%¹09.6%?n=6?,and the RSDs range is 4.8%⁵.1%?n=6?.The experimental results showed that the contents of two kinds of saponins in three Shenmai injections made by different manufacturers are 15.98⁴1.15?g/mL?DT-13?and 1.18⁴.54?g/mL?Ophiopogon saponin D?repectively.The content of DT-13 is higher than that of Ophiopogon saponin D,but the contents of the two in different injections are quite different.In summary,Rg₆ and DT-13 are newly discovered from Shenmai injection,and the determination method of seven rare ginsenosides is established for the first time.The sources of these rare ginsenosides are also investigated.The results of this study not only provide a new sample pretreatment method for the determination of the chemical constituents of rare ginsenosides and Ophiopogon saponins in Shenmai injection,but also provide a new scientific basis for improving the quality standards and secondary development of Shenmai injection.