The scape of this protocol lies in the proceeding of the Tibetan medicines. This start aims to the optimize and standardize the processing technology of the zanba stir fried tiebangchui. The combination of the Box-Behnken response surface and the critical method is commonly used to optimize processing technology to acquire the optimize precising protocol.
The optimized the processing conditions of a two centimeter tiebangchui slice thickness three times more than modern tiebangchui a processing temperature of 125 degrees Celsius and 60 minutes of stir frying by qualifying the specific processing parameters this method will benefit the large scale production and the quality control of Zanba stir fried tiebangchui. The standard Zanba stir fried processing technology of tiebangchui can provide useful information and guidance for the processing of other poisonous ethnic medicines. The optimized process is stable and feasible, laying a certain foundation for further investigation of tiebangchui products and related Chinese medicine preparations.
Our laboratory will pay more attention to cyto chemical investigation to obtain the target quality control compliance. In addition, the pharmacological comparison overall on zanba stir fry tiebangchui is unclear the detoxification and evaluation of the efficacy reservation effects in animal models will be the next objectives. To begin a single factor experiment to compare tiebangchui or TBC slice thickness for processing technology optimization.
Prepare five test groups each having 30 grams of TBC, but different slice thickness. Then weigh 90 grams of zanba which is three times as much as that of the TBC In each group. Set the temperature and the time of the automatic stir fry machine to 140 degrees Celsius and 40 minutes respectively.
After the machine has heated up to the set temperature, add the weight amounts of TBC and zanba into the machine. Then using a high speed smashing machine, grind the zanba stir fried TBC into powder. Next, weigh two grams of zanba stir fried TBC powder in a conical flask to the sample in the flask, add three milliliters of ammonia solution followed by 50 milliliters of a mixture of isopropyl alcohol and ethyl acetate.
Having a volume ratio of one is to one. After recording the weight of the conical flask with the contents ultrasonic the TBC sample for 30 minutes at 220 volts and 40 kilohertz, maintaining a temperature below 25 degrees Celsius. Weigh the conical flask with the sample once again after the ultrasonic extraction.
Makeup for the lost weight by adding the isopropyl alcohol and ethyl acetate mixture mentioned previously, and filter the sample solution. Evaporate 25 milliliters of the filtrate to dryness. Using a rotary evaporator at 40 degrees Celsius, then dissolve the residue in five milliliters of a solution of 0.05%hydrochloric acid in methanol.
Filter the solution through a 0.2 micron syringe filter and analyze it by performing high performance liquid chromatography or HPLC. Calculate the contents of the MDA and DDAs in different processing products according to the standard curve. Calculate the comprehensive score based on the results via the critic method.
Following the method demonstrated above. Compare the amounts of ZBA processing temperatures and times for processing technology optimization. The results of the single factor tests revealed that the comprehensive scores of zanba stir fried TBC were highest When the sliced thickness is two centimeters, the amount of zanba is three times, the processing temperature is 120 degrees Celsius and the processing time is 60 minutes.
The results revealed the range of slicing thickness, amount of zanba, processing temperature and processing time to be used for future experiments. To perform the box bank in response surface design for optimization of zanba stir-fried tiebangchui or TBC processing technology. Use the range of thickness, the amount of zanba the processing temperature, and the processing time determined by preliminary experiments using single factor tests.
On the homepage click on new design, and in the left panel of the design page, click on response surface. Then click Box-Behnken and set the parameters for the four factors in the table. Click on next, set the response names and click on finish.
To generate the response surface design. Complete the experiment based on the 29 scenarios designed for the response surface. Using high performance liquid chromatography or HPLC.
Record the peak areas and determine the quality of the MDA and DDAs in the zanba Stir fried TBC test samples. Then input the comprehensive scores obtained via the critic method for the 29 trials into the computer and analyze it using the referenced software. To perform the statistical validation of the polynomial equations in the left navigation pane under analysis plus, click on R1 and then click on start analysis.
In the configure window, click on ANOVA in the top menu and observe the results table displaying variance analysis. Next in the top menu, click on Model Graphs and then 3D Surface to obtain the response surface plots reflecting the effects of processing parameters on the synthetic scores. Go to the left navigation pain and under Optimization, click on Numerical to validate the response surface model in triplicate under the predicted optimum conditions.
Then in the top menu, click on Solutions. Observe the predicted optimal conditions to verify the stability of the processing technology. The response surfaces and contour plots demonstrated the changes in synthetic scores as a function of four variables on the basis of the experimental results.
The adjusted optimal processing conditions were a slice thickness of two centimeters, three times more ZBA than TBC, a processing temperature of 125 degrees Celsius, and a processing time of 60 minutes. The reliability of the model was proven through three tests that were conducted according to the obtained processing parameters.