This work was financially supported by the National Natural Science Foundation of China (No. 82130113), the China Postdoctoral Science Foundation (No. 2021MD703800), the Science Foundation for Youths of Science &#38; Technology Department of Sichuan Province (No. 2022NSFSC1449), and the &#34;Xinglin Scholars&#34; Research Promotion Program of Chengdu University of Traditional Chinese Medicine (No. BSH2021009).

The Zanba-stir-fried TBC processing method was optimized and standardized by CRITIC combined with the Box-Behnken response surface method. Benzoylaconitine, aconitine, and 3-deoxyaconitine were used as evaluation indexes during this procedure.
1. Sample solution preparation
<ol>
	<li>Prepare the reference substance stock solution. Weigh precisely 9.94 mg of benzoylaconitine, 8.49 mg of aconitine, and 6.25 mg of 3-deoxyaconitine (Table of Materials) on an ele...

A Method for Zanba-Stir-Fried Tiebangchui Process Optimization

<ol>
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TBC is an important Tibetan medicine with the effects of dispelling cold and relieving pain. It has been mostly used to treat traumatic injury and rheumatic arthralgia in China for thousands of years24,25,26. Diterpenoid alkaloids are both active and toxic ingredients of TBC27,28,29. The main toxic effects of the aconitum alkaloids of T...

The dried root of Aconitum pendulum Busch and A. flavum Hand.-Mazz., one of the most famous Tibetan medicines, is called Tiebangchui (TBC) in Chinese1,2. The dried roots of TBC are helpful in dispelling cold and wind, reducing pain, and calming shock. It was recorded in the first volume of "Drug Standards (Tibetan Medicine) of the Ministry of Health of the People's Republic of China," which states that the dried roots of TBC are commonly used to treat rheumatoid arthritis, bruises, and other cold diseases3. However, the clinical therapeutic dose of TBC is similar to its toxic dose, and incidents of poisoning or death have been frequently reported due to improper use4. Therefore, reducing the toxicity and preserving the efficacy of TBC has become a research hot spot over the years.
In Tibetan medicine, processing is one of the most effective methods to attenuate the toxicity of TBC. According to "Processing specification of Tibetan medicine of Qinghai Province (2010)", the original herbs (TBC) should be placed in an iron pot and stir-fried with Zanba until the Zanba turns yellow, after which Zanba is removed and the herbs are dried in air5,6. However, no specific process parameters have been documented, which makes controlling the processing technology and the quality of Zanba-stir-fried TBC difficult. The CRITIC method is an objective weight method that can avoid fuzzification and subjectivity, and enhance the objectivity of weighing7. The Box-Behnken response surface method can directly reflect the interaction between each factor through polynomial fitting8. The combination of the Box-Behnken response surface and CRITIC method is commonly used to optimize processing technology to acquire the optimized processing protocol9,10. In this paper, a monoester-diterpenoid alkaloid (MDA) (benzoylaconitine) and two diester-diterpenoid alkaloids (DDAs) (aconitine, 3-deoxyaconitine) were used as evaluation indexes. CRITIC combined with the Box-Behnken response surface method was applied to optimize the processing technology of Zanba-stir-fried TBC and establish a standard processing method for clinical safe use.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3-Deoxyaconitine</td>
			<td>Chengdu Desite Biotechnology Co., Ltd.</td>
			<td>DST221109-033</td>
			<td></td>
		</tr>
		<tr>
			<td>Aconitine</td>
			<td>Chengdu Desite Biotechnology Co., Ltd.</td>
			<td>DSTDW000602</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium acetate</td>
			<td>Tianjin Kermel Chemical Reagent Co., Ltd</td>
			<td>Chromatographic grade</td>
			<td></td>
		</tr>
		<tr>
			<td>Benzoylaconitine</td>
			<td>Chengdu Desite Biotechnology Co., Ltd.</td>
			<td>DSTDB005502</td>
			<td></td>
		</tr>
		<tr>
			<td>Design-Expert software</td>
			<td>Stat-Ease, Inc., Minneapolis, MN, USA</td>
			<td>version 13.0</td>
			<td></td>
		</tr>
		<tr>
			<td>Electronic analytical balance</td>
			<td>Shanghai Liangping Instruments Co., Ltd.</td>
			<td>FA1004</td>
			<td></td>
		</tr>
		<tr>
			<td>High performance liquid chromatography</td>
			<td>SHIMADZU Co., Ltd.</td>
			<td>LC-20A</td>
			<td></td>
		</tr>
		<tr>
			<td>High-speed smashing machine</td>
			<td>Beijing Zhongxing Weiye Instrument Co., Ltd.</td>
			<td>FW-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Millipore filter</td>
			<td>Tianjin Jinteng Experimental Equipment Co., Ltd</td>
			<td>&#966;13 0.22 Nylon66</td>
			<td></td>
		</tr>
		<tr>
			<td>stir-Fry machine</td>
			<td>Changzhou Maisi Machinery Co., Ltd</td>
			<td>Type 5</td>
			<td></td>
		</tr>
		<tr>
			<td>Tiebangchui</td>
			<td>Gannan Baicao Biotechnology Development Co., Ltd</td>
			<td>20211012</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultra pure water systemic</td>
			<td>RephiLe Bioscience, Ltd.</td>
			<td>Genie G</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic cleansing machine</td>
			<td>Ningbo Xinyi Ultrasonic Equipment Co., Ltd</td>
			<td>SB2200</td>
			<td></td>
		</tr>
		<tr>
			<td>Zanba</td>
			<td>27 Chuanzang Road, Ganzi County</td>
			<td>-</td>
			<td></td>
		</tr>
	</tbody>
</table>

optimization of processing technology for tiebangchui with zanba based on critic combined with box-behnken response surface method

The dried root of Aconitum pendulum Busch., called Tiebangchui (TBC) in Chinese, is one of the most famous Tibetan medicines. It is a widely used herb in northwest China. However, many cases of poisoning have occurred because of TBC's intense toxicity and because its therapeutic and toxic doses are similar. Therefore, finding a safe and effective method to reduce its toxicity is an urgent task. A search through the Tibetan medicine classics shows that the processing method of TBC stir-fried with Zanba was recorded in the "Processing specification of Tibetan medicine of Qinghai Province (2010)". However, the specific processing parameters are not yet clear. Thus, this study aims to optimize and standardize the processing technology of Zanba-stir-fried TBC.
First, a single-factor experiment was conducted on four factors: the slice thickness of TBC, amount of Zanba, processing temperature, and time. With monoester and diester alkaloid contents in Zanba-stir-fried TBC as indexes, CRITIC combined with the Box-Behnken response surface method was used to optimize the processing technology of Zanba-stir-fried TBC. The optimized processing conditions of Zanba-stir-fried TBC were a TBC slice thickness of 2 cm, three times more Zanba than TBC, a processing temperature of 125 &#176;C, and 60 min of stir-frying. This study determined the optimized and standard processing conditions for the usage of Zanba-stir-fried TBC, thus providing an experimental basis for the safe clinical use and industrial production of Zanba-stir-fried TBC.

Author Spotlight: Optimization of Processing Technology for Tiebangchui with Zanba Based on CRITIC Combined with Box-Behnken Response Surface Method  

Optimization of Processing Technology for Tiebangchui with Zanba Based on CRITIC Combined with Box-Behnken Response Surface Method

The scope of this protocol lies in the proceeding of the Tibetan medicines. This staff aims to the optimize and standardize the processing technology of Zanba-stir-fried Tiebangchui. The combination of the Box-Behnken response surface and the CRITIC method is commonly used to optimize proceeding technology to acquire the optimized proceeding protocol.The optimized processing conditions of two centimeter Tiebangchui slice thickness three times more Zanba than Tiebangchui, a processing temperature of 125 degrees C and the 60 metres of stir-frying. By quantifying the specific processing parameters, this method will benefit the large-scale production and the quantity control of Zanba-stir-fried Tiebangchui. The standard Zanba-stir-fried processing technology of Tiebangchui can provide useful information and the guidance for the processing of other poisonous ethnical 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 cytochemical investigation to obtain the targeted quantity control compliance. In addition, the pharmacological comparison overall in Zanba-stir-fried Tiebangchui is unclear.Detoxification and evaluation of the efficacy, reservation effects in animal models will be the next objectives.

In this study, the elution gradient used had a good resolution (Figure 1) for the three index components in Zanba-stir-fried TBC, as determined after repeated debugging. The three index components in Zanba-stir-fried TBC had a good linear relationship within a specific concentration range (Table 2). The precision (Table 5), stability (Table 6), repeatability (Table 7), and sample recovery (Table 8) of Zanba-...

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The present protocol describes an efficient and standard detoxification processing method for Zanba-stir-fried Tiebangchui using CRITIC combined with the Box-Behnken response surface method.

The dried root of Aconitum pendulum Busch., called Tiebangchui (TBC) in Chinese, is one of the most famous Tibetan medicines. It is a widely used herb in ...

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JoVE Journal

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Preparing Zanba-Stir-Fried Tiebangchui (TBC) Sample for Processing Technology Optimization

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 weighed 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, ultrasonicate 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.Make up 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 Zanba, 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.

Response Surface Methodology (RSM) for Processing Technology Optimization of Zanba-Stir-Fried Tiebangchui (TBC)

To perform the Box-Behnken in response surface design for optimization of Zanba-Stir-Fried Tiebangchui or TBC processing technology. Use the range of sliced 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 pane 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 Zanba 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.

667 Views.  Chengdu University of Traditional Chinese Medicine.  The present protocol describes an efficient and standard detoxification processing method for Zanba-stir-fried Tiebangchui using CRITIC combined with the Box-Behnken response surface method.

A Method for Zanba-Stir-Fried Tiebangchui Process Optimization

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