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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