A Consistency Evaluation Of Preliminary Study Of Traditional Decoction Method And Ten Function Decoction Machine Method Of Gegen Qinlian Decoction(GQD)

Objective:The classic prescription-Gegen Qilian Decoction(GQD)was taken as the representative.The differences between the traditional decoction method and the decoction of ten-function decocting machine method were compared by the system,which provided the experimental basis and evaluation basis for further promotion of the modern decoction of ten-function decocting machine for the application of traditional Chinese medicine prescriptions.Methods:1.The multi-components such as baicalin,puerarin,berberine hydrochloride and glycyrrhizin were used as indicators to establish a high-performance liquid chromatography method for“GQD”.2.The relationship between the number of viable bacteria of Staphylococcus aureus,Escherichia coli,Shigella flexneri and absorbance was determined.A total colony counting method for Staphylococcus aureus,Escherichia coli,and Shigella flexneri in the bacterial solution was established.At the same time,the K-B paper method and the micro broth dilution method were applied.The antibacterial effect evaluation method of Staphylococcus aureus,Escherichia coli,Shigella flexneri,the minimum inhibitory concentration and the minimum bactericidal concentration was established as the index of "GQD".3.The differences between traditional decoction and decoction of ten-function decocting machine method for GQD in boiling time,odor and appearance of the liquid,manpower input,decoction rate,index component content,and in vitro antibacterial efficacy were compared.4.The multi-indicator component was combined with the antibacterial effect evaluation to investigate the soaking time,water addition,boiling time,boiling pressure and boiling time of the "GQD" by the modern decoction of ten-function decocting machine process.Results:1.Chromatographic conditions:Agilent ZORBAX Eclipse SB-C18 column(4.6 mm X 250 mm,5 ?m);Mobile phase:10 mmol·L-1 ammonium formate(pH value=3.0,adjusted by formic acid)(A)acetonitrile(B)gradient elution;Flow rate:1.0 mL·min-1;Detection of dual wavelength:270 and 360 nm;Column temperature:25 ?;Injection volume:20 ?L.Complete separation of the various indicator components can be achieved and R was greater than 1.5.Precision,repeatability,stability,and relative standard deviation(RSD)were both<2.0%,indicating that the method is accurate and feasible.2.The linear regression equation of Staphylococcus aureus was y=8.770x-0.376(R2=0.992),and the total number of bacteria in the range of 1.2×108?8.0×108 was linear.The linear regression equation of Escherichia coli wasy=9.083x-0.736(R2=0.988),and the total number of bacteria was linear in the range of 1.4×108?7.8×108.The linear regression equation of Shigella flexneri was y=8.671x-0.076(R2=0.994),and the total number of bacteria in the range of 1.6×108?7.9×108 was linear.3.Compared with the traditional decoction method,the ten-function decocting machine method for GQD had a short boiling time(68min),less sediment,smaller particle size and less viewing time(2 times),no boiling,gelatinization.And the smell and color of the liquid were basically the same(gas thick,bitter,brown).4.The best single factor of "GQD" was:soak for 60min;20 times water was added and decocting for 90 min at normal pressure for the first time,and 10 times water was added for 15min at normal pressure for the second time.The best process of the modern decoction of ten-function decocting machine method was as follows:the medicinal material was immersed for 45 minutes;13 times water was added at normal pressure and decocting for 80 minutes for the first time;6.5 times water was added at normal pressure and decocting for 10 minutes for the second time.5.Comparison of content determination results:The content of baicalin,puerarin,berberine hydrochloride and glycyrrhizin in traditional decoction method and decoction of modern ten-function decocting machine method were determined for each 1 ml of GQD sample solution(0.65 g crude drug/ml).The active ingredients of the two were basically the same.6.Comparison of antibacterial results:The minimum inhibitory concentration and minimum bactericidal concentration of the modern ten-function decocting machine method on Staphylococcus aureus,Escherichia coli,and Shigella flexneri were lower than the traditional decoction method,that is,the effect of suppressing(killing)bacteria was also superior to the traditional method.Conclusions:The decoction of ten-function decocting machine methods for GQD saves time,labor with high efficiency and good effect,and is superior to the traditional decoction method.It is worthy of further promotion and application,and can provide a basis for the modern ten-function decocting machine for Chinese medicine prescription.