As the company used to debate on medicine with toxic effect is proceeding Tiebangchui for toxicity attenuation is extremely important for clinical application. The optimize proceeding technology of the Tiebangchui with a highland barley wine is simple, visible, and stable. which can provide this variance for industrial production.
Box-Behnken design combined with the entropy method allows the inter-table observation of optimized conditions and has been widely used in chemistry, pharmacy, and food precision. Demonstrating the procedure will be Liqiong Yu, a master student from my laboratory. To begin, take 500 grams of black highland barley rice, and add water to five times the amount of rice.
Cook the rice for approximately two hours, or until the remaining water is absorbed. Once cooked, pour the rice out, and wait until the temperature falls to 37 degrees Celsius. Add four grams of jiuqu to the rice, and mix well.
Seal the can, and wrap the container with cotton wool. Then, let it stew for seven days. Add 300 milliliters of water on the seventh day, and reseal the container.
On the eighth day, remove the wine. And replace it with 300 milliliters of water. Seal the container properly, and allow it to ferment for one day.
Pool the collected wine into a container. Next, accurately weigh the Tiebangchui, or TBC, into a container, and add highland barley wine to soak the mixture for one day. The following day, dry it in a constant-temperature electric drying oven.
To prepare the test sample solution, accurately weigh two grams of the TBC processed product powder in a conical flask, and add 40%ammonia solution. Next, perform ultrasound-assisted extraction for 30 minutes, with a 50-milliliter mixture of isopropanol and ethyl acetate in an equal ratio. To perform the highland barley wine addition test, set up five groups, each with 30 grams of 1.0 centimeter thick TBC.
Add varied amounts of Hyland barley wine to each group containing the TBC, and soak the slices for 12 hours. To perform the soaking time test, set up five groups, each with 30 grams of 1.0 centimeter thick TBC. Soak the TBC with highland barley wine at five times the amount of TBC in all the groups, with different time intervals.
To perform the slicing thickness test, set up five groups with 30 grams of TBC at varied sliced thicknesses. Soak the TBC for 24 hours with highland barley wine at five times the amount of TBC in all the groups. As previously demonstrated, prepare the test sample solution for each test group, with the processed products.
Next, determine the peak area of each sample by HPLC. Use the standard curve to estimate the amount of monoester-diterpenoid alkaloids, or MDAs, and diester-diterpenoid alkaloids, or DDAs. Use the content of MDAs and the total content of DDAs as evaluation indicators to determine the weight coefficient of each evaluation index, and the comprehensive scoring by the entropy method.
Use equation two to standardize the content of MDAs, where XIJ is the value of the Jth indicator of the Ith sample, and use equation three to standardize the total content of the DDAs. Calculate the probability of the Jth trial under the Ith evaluation indicator PIJ, using equation four, and use equation five to calculate the information entropy HJ.Use equation six to calculate the indicator weights, WJ, and equation seven to calculate the comprehensive scoring of the indicators. open the software on the computer.
Click New Design, Response Surface, and then select Box-Behnken Design. Input the number of influencing factors, and the level of information of the experiments in the study, and then click Continue. Input the number of response values, and click Finish to complete the process.
Then, set the comprehensive scoring by equations two to seven as the response. Next, process the TBC with highland barley wine, according to the design results, and complete the experiment based on the 17 scenarios designed for the response surface. Prepare the sample solutions as previously demonstrated, and calculate the total content of the MDAs and DDAs, using HPLC.
Click Analyze to analyze the data and model information. Perform statistical validation by ANOVA, in the top menu, and observe the results table. Click Optimization, and then Numerical, to view the predicted optimal process conditions.
Click on Analysis, Model Graphs, and then 3D surface to view the response surface analysis plotted as three-dimensional plots by the software. The precision test results assessed by the relative standard deviation of the peak areas of benzoylaconine, aconitine, 3-deoxyaconitine, and 3-acetylaconitine, indicated that the precision of the instrument was good. The stability test suggested that all the sample solutions were stable for 24 hours.
And the repeatability test results showed that the repeatability of this method was significant. The recovery experiment results indicated average recovery rates as 99.7%for benzoylaconine, 100.84%for aconitine, 103.27%for 3-deoxyaconitine, and 100.92%for 3-acetylaconitine. The single-factor test of TBC processed with highland barley wine, revealed that the amount of highland barley wine was five times that of TBC, the soaking time was 36 hours, and the slicing thickness was 1.0 centimeters.
The experimental design and results of the response surface model are tabulated here. The order of the factors by the strength of the effects was established, where it was higher for highland barley wine addition, followed by slice thickness of medicinal herbs, and then the soaking time. It's essential to have three periods of experiment, for each condition of a single factor to improve the accuracy of subsequent results.
The optimization polarization technology can provide information and guidelines for studying toxic intonation to produce other toxic ethical medicines.
