Based On Conventional Extraction Method And Optimization Of Ultrahigh Pressure Extraction Process Of Jinqiao Reduqing

Jinqiao Reduqing Granule takes genuine medicinal materials as raw material and is composed of Folium isatidis, Lonicera japonica, Forsythia suspense, plaster stone, Fineleaf Schizonepeta Herb, Peppermint, Uncaria tomentosa, Periostracum cicadae, Scrophularia ningpoensis, Reed Rhizome, Common hogfenneI root, ageratum, medicated leaven, Lophatherum gracile and Licorice. And these medicinal materials determine the activities on relieving exterior syndrome by inducing mild diaphoresis, clearing heat and removing toxicity, relieving sore throat and cough, etc. As for clinical application, it is usually used to treat upper respiratory tract infection and acute upper respiratory tract infection, such as common cold, fever, seasonal febrile diseases, headache, sore throat, tonsillitis, and so on. It obtained the "hospital preparation approval" in1991, and began to be produced and used in our hospital. During the period of clinical application, Jinqiao Reduqing Granule’s side effect had not been observed, and no adverse events reported so far. Therefore, it is necessary to do some in-depth study of Jinqiao Reduqing Granule. The aim of this study is to explore the extraction process of Jinqiao Reduqing Granule, and to provide the experimental basis for fully ensuring the efficacy of Jinqiao Reduqing Granule.On the basis of previous studies, the problem of extraction process (12,8,8times amount of water;1.5,1,1h,3times) of Jinqiao Reduqing Granul is mainly low extraction yield of chlorogenic acid and forsythiaside. First, chlorogenic acid is formed by caffeic acid and quinic acid, and its molecular structure has three unstable parts, including ester bond, unsaturated double bond and polyphenol; and forsythiaside is benzyl alcohol coffee acid compounds, has ester bond and glycosidic bond, is easy to completely decompose into coffee acid, D-glucose, L-rhamnose and asecondary glycoside under high temperature conditions. Both of them are heat sensitive components, and cannot accept the high temperature heating for a long time. Second, alkaloids are the main active ingredients of Uncaria tomentosa, acid-base reaction occurs easily between organic acid (such as, chlorogenic acid) and alkaloids.At present, there are many defects for traditional methods (such as reflux extraction, soxhlet extraction, cold soak extraction, percolation extraction, steam distillation), including big loss, high energy consumption, high solvent consumption, long period, massed operations, low extraction rate, and the active ingredient is not clear. It is urgent to find a fast and effective extraction method of high activity、stable quality for effective components. And ultrahigh pressure extraction is the emergence of a new kind of low temperature extraction technology in recent years, its operating principle is first to mix crushed herbs and extraction medium in airtight containers, increase rapidly the pressure up to100MPa, hold pressure (1-10min), and quickly pressure relief. In the ultrahigh pressure extraction process, high pressure greatly accelerates infiltration and solute diffusion process, makes the pores full of solvent, improves the mass transfer velocity of effective components. Its characteristics are higher extraction yield, lower extraction temperature, shorter extraction time, less power consumption, less impurity content, eco-friendly and so on, especially is suitable for extracting heat-sensitive components.In the present work, we plan to solve the problem of low extraction yield of chlorogenic acid and forsythiaside for Jinqiao Reduqing Granule. First, to explore the causes of low retention rates of effective components. Meanwhile, we optimized the ultrahigh pressure extraction process of effective constituents from Jinqiao Reduqing granule, improved the extraction yield of the main effective components, providing the experimental basis for fully ensuring the efficacy of Jinqiao Reduqing Granule.Objective:To optimize the UPE process of chlorogenic acid and forsythiaside from Jinqiao Reduqing granule, obtaining the higher extraction rates. And, to solve the problem of the preparation technique of Jinqiao Reduqing Granule.Methods:1To explore the reason for low extraction yield of chlorogenic acid and forsythosideThe response variable was the extraction yield of chlorogenic acid or forsythiaside. We compared the extraction yield of chlorogenic acid from Lonicera japonica or forsythiaside from Forsythia suspense of water decoction method with warm immersion method. Determination of the extraction yield of chlorogenic acid of water decoction method from Lonicera japonica, Uncaria rhynchophylla, simultaneously extracting Lonicera japonica and Uncaria rhynchophylla, we investigated the effects of pH of extraction solvent and Uncaria rhynchophylla on the content of chlorogenic acid of Lonicera japonica. Finally, the results were analyzed by SPSS13.0software.2Optimization the UPE process of chlorogenic acid from Lonicera japonica and forsythoside from Forsythia suspensaOn the basis of single factor screening, a three-factor and three-level Box-Behnken experimental design was employed, with ethanol concentration (X1), liquid/material ratio(X2) and extraction pressure (X3) as independent variables. The response variable was the extraction yield of chlorogenic acid and forsythiaside.3Optimization the UPE process of effective constituents from Jinqiao Reduqing granuleOn the basis of single factor screening, a three-factor and three-level Box-Behnken experimental design was employed, with solvent fold (X1), extraction pressure (X2) and extraction time (X3) as independent variables. Dependent variables were transformed into desirabilities mathematically by Hassan’s method. Data of overall desirabilities were fitted to a second order polynomial equation, through which three dimensional response surface graphs were produced. Optimum experimental conditions were selected from the stationary point of the response surfaces. The extraction results of UPE were compared with heat reflux and water decoction extraction.Results:1To explore the reason for low extraction yield of chlorogenic acid and forsythosideDetermination of the extraction yield of chlorogenic acid from Lonicera japonica or forsythiaside from Forsythia suspense, the emergence of new chromatographic peaks of HPLC chromatograms demonstrated chlorogenic acid and forsythiaside might degrade during the decoction process. And the longer decoction time of Lonicera japonica was, the lower extraction yield of chlorogenic acid was. Higher extraction rate of chlorogenic acid can be obtained in the extraction solvent of pH3.0compared with the extraction solvent of other values of pH(P<0.05). However, simultaneously extracting Lonicera japonica and Uncaria rhynchophylla for decocting time more than an hour, preliminary results had showed Uncaria rhynchophylla decreased the extraction yield of chlorogenic acid from Lonicera japonica.2Optimiztion the UPE process of chlorogenic acid from Lonicera japonica and forsythoside from Forsythia suspensaThe optimal conditions of UPE for chlorogenic acid and forsythiaside were that the ratio of solvent to material (mL·g-1) was75and70, the extraction pressure was100and151MPa, and the extraction time was28and114second, respectively. With this extracting process, the extraction yield of chlorogenic acid and forsythiaside was25.43and13.15mg·g-1respectively. It was11.35%or8.64%higher than reflux extraction for chlorogenic acid or forsythiaside,12.16%or6.52%to ultrasonic extraction and15.98%or10.43%to water decoction method. With this extracting process, the extraction yield of chlorogenic acid was25.2937,25.6423,25.4527mg·g-1, and forsythiaside was13.1351,13.1367,13.1817mg·g-1.3Optimization the UPE process of effective constituents from Jinqiao Reduqing granuleThe optimum conditions of UPE were as follows:the materials were extracted for38s with24times amount of water, the extraction pressure was112.1MPa, and the extraction was performed for one time. The results of significant test showed that X1, X2, X3, X12of model were different significantly. Regression coefficient of binomial fitting complex model was as high as0.9878, and bias between observed and predicted values was5.27%. And, the model can be used to analyze and predict. The extraction results of UPE were different significantly, compared with heat reflux、 water decoction method (P<0.05or P<0.01).Conclusion:The current conventional extraction process (water decoction method) was used for Jinqiao Reduqing, but the extraction yields of chlorogenic acid and forsythiaside are low. Chlorogenic acid and forsythiaside might degrade during the decoction process, higher extraction rate of chlorogenic acid can be obtained in the extraction solvent of pH3.0, and simultaneously extracting Lonicera japonica and Uncaria rhynchophylla led to lower the extraction yield of chlorogenic acid. However, rhynchophylline did not affect the content of chlorogenic acid. The UPE can improve extraction yield of chlorogenic acid and forsythiaside from Jinqiao Reduqing granule. As a novel extraction technology for Chinese herbal medicine, the UPE procedure has simpler extraction process, shorter extraction time, lower extraction temperature. And its extended application can be expected to offer a new method for extracting effective components of traditional Chinese herbal medicine.