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).

In this study, the processing technology of TBC processed with highland barley wine was optimized with a Box-Behnken design combined with the entropy method. DDA and MDA content were used as evaluation indicators, and the weight coefficient of each evaluation index was determined by the entropy method.
1. Experimental preparation
<ol>
	<li>Prepare highland barley wine15.
	<ol>
		<li>Take 500.00 g of black highland barley rice and add five times the am...

<ol>
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C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Box-Behnken+design:+an+alternative+for+the+optimization+of+analytical+methods.">Box-Behnken design: an alternative for the optimization of analytical methods.</a> Analytica Chimica Acta. 597 (2), 179-186 (2007).</li><li>Dong, R., Lu, Y., Wang, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+process+optimization+of+vinegar+roasting+of+Bupleurum+chinense+by+entropy+weight+method+combined+with+Box-Behnken+response+surface+method+and+its+protective+effect+on+mice+liver+injury.">The process optimization of vinegar roasting of Bupleurum chinense by entropy weight method combined with Box-Behnken response surface method and its protective effect on mice liver injury.</a> Science and Technology of Food Industry. 42 (23), 209-217 (2021).</li><li>Li, W. 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The authors have no conflicts of interest to disclose.

As a commonly used Tibetan medicine with toxic effects, the toxicity-attenuating effect of processing is extremely important for TBC&#39;s clinical application25. In this study, the processing technology of TBC processed with highland barley wine was optimized. By reviewing the main active ingredients and relating the pharmacological effects of TBC, we found that TBC alkaloids have anti-inflammatory and analgesic effects and can be used to treat rheumatoid arthritis. In this study, a moderate toxi...

Tiebangchui (TBC), the dried root of Aconitum pendulum Busch, is a well-known Tibetan medicine and was initially recorded in the classic Tibetan medical book &#34;Four Medical Tantra&#34;1,2. According to &#34;Drug Standards of the Ministry of Health of the People&#39;s Republic of China (Tibetan Medicine)&#34;, TBC is effective in expelling cold, relieving pain, dispelling wind, and calming shock, and is commonly used to treat rheumatoid arthritis in clinics3,4,5.
TBC mainly contains alkaloids, including highly toxic diester-diterpenoid alkaloids (DDAs), and the moderately toxic monoester-diterpenoid alkaloids (MDAs)6,7,8. These chemical components are active ingredients with medicinal effects but are toxic. One of the most famous active and toxic ingredients, aconitine, causes poisoning when it exceeds 1 mg9. Therefore, improper or excessive use of TBC might result in poisoning and even death, and the toxicity attenuation and efficacy reservation of TBC is crucial for its safe clinical application10,11.
Processing is an effective method for detoxifying TBC. According to ancient Tibetan medicine books, processing with highland barley wine is an efficient way to attenuate toxicity and preserve the efficacy of TBC. TBC is soaked in highland barley wine, stored for one night, dried, and added to medicines12. However, the specific processing technology and potential influencing factors are rarely reported, and the traditional processing process often relies on experience and lacks standardized methods. Hence, modern scientific and technological methods for optimizing and standardizing the processing process are needed.
The Box-Behnken design method is used in investigating interactions among different factors and their influence on comprehensive scoring through quadratic polynomial fitting. This design allows the intuitive observation of optimal conditions and has been widely used in the field of pharmacy13. For example, the Box-Behnken design method, based on the entropy method, successfully optimized the processing technology of stir-frying with vinegar of Curcuma Longa Radix14. In this study, the Box-Behnken response surface experimental design combined with the entropy method was used in optimizing the processing technology of TBC processed with highland barley wine. The optimized processing technology is expected to ensure quality control and safe clinical use.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Aconitine</td>
			<td>Chengdu Push Biotechnology Co.,Ltd</td>
			<td>PS000905</td>
			<td></td>
		</tr>
		<tr>
			<td>3-Acetylaconitine</td>
			<td>Chengdu Push Biotechnology Co.,Ltd</td>
			<td>PS010552</td>
			<td></td>
		</tr>
		<tr>
			<td>3-Deoxyaconitine</td>
			<td>Chengdu Push Biotechnology Co.,Ltd</td>
			<td>PS011258</td>
			<td></td>
		</tr>
		<tr>
			<td>Benzoylaconine</td>
			<td>Chengdu Push Biotechnology Co.,Ltd</td>
			<td>PS010300</td>
			<td></td>
		</tr>
		<tr>
			<td>Circulating water vacuum pump</td>
			<td>Gongyi City Yuhua Instrument Co., Ltd</td>
			<td>SHZ-DIII</td>
			<td></td>
		</tr>
		<tr>
			<td>Design-Expert&#160;</td>
			<td>State-East Corporation</td>
			<td>8.0.6</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric constant temperature drying oven</td>
			<td>Shanghai Yuejin Medical Equipment Co., Ltd</td>
			<td>101-3-BS</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 Enterprise Management (China) Co., Ltd</td>
			<td>shimadzu 2030</td>
			<td></td>
		</tr>
		<tr>
			<td>Highland barley rice</td>
			<td>Kangding City, Ganzi Tibetan Autonomous Prefecture, Sichuan Province</td>
			<td>20221015</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>Rotary evaporator</td>
			<td>Shanghai Yarong Biochemical Instrument Factory</td>
			<td>RE-2000A</td>
			<td></td>
		</tr>
		<tr>
			<td>Starter of liquor-making</td>
			<td>Angel Yeast CO., Ltd</td>
			<td>BJ22-104</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultra pure water systemic</td>
			<td>Merck Millipore Ltd.</td>
			<td>Milli-Q</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic cleansing machine</td>
			<td>Ningbo Xinyi Ultrasonic Equipment Co., Ltd</td>
			<td>SB-8200 DTS</td>
			<td></td>
		</tr>
	</tbody>
</table>

optimization of processing of tiebangchui with highland barley wine based on the box-behnken design combined with the entropy method

The processing of toxic ethnomedicines is of great significance for their safe clinical application. Thus, the limitations of traditional processing should be addressed, and the processing method of ethnomedicines should be standardized using modern research methods. In this study, the processing technology of a commonly used Tibetan medicine Tiebangchui (TBC), the dried root of Aconitum pendulum&#160;Busch, processed with highland barley wine was optimized. Diester-diterpenoid alkaloid (DDA) (aconitine, 3-deoxyaconitine, 3-acetylaconitine) and monoester-diterpenoid alkaloid (MDA) (benzoylaconine) content were used as evaluation indicators, and the weight coefficient of each evaluation index was determined by the entropy method.
The single factor test and Box-Behnken design were used in investigating the influence of the ratio between highland barley wine and TBC, slice thickness of TBC, and processing time. Comprehensive scoring was performed according to the objective weight of each index determined by the entropy method. The optimal processing conditions of TBC with highland barley wine were as follows: the amount of highland barley wine is five times that of TBC, a soaking time of 24 h, and a TBC thickness of 1.5 cm. The results showed that the relative standard deviation between the verification test and predicted value was less than 2.55% and the optimized processing technology of TBC processed with highland barley wine is simple, feasible, and stable, and so can provide a reference for industrial production.

In this study, the precision, stability, repeatability, and sample recovery of TBC indicated that the method is feasible. The four index components in TBC had a good linear relationship within a specific concentration range. Typical chromatograms are shown in Figure 1. The precision test results (Table 8) showed that the relative standard deviation (RSD) of the peak areas were 2.56%, 1.49%, and 2.03% for benzoylaconine, aconitine, and 3-deoxyaconitine, respectively, and 0.21...

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Optimization of Processing of Tiebangchui with Highland Barley Wine Based on the Box-Behnken Design Combined with the Entropy Method

The present protocol describes an efficient method for optimization of the processing technology of Tiebangchui processed with highland barley wine based on a Box-Behnken design response surface combined with the entropy method.

The processing of toxic ethnomedicines is of great significance for their safe clinical application. Thus, the limitations of traditional processing ...

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.

optimization-processing-tiebangchui-with-highland-barley-wine-based

Research

JoVE Journal

Medicine

525 조회수.  Chengdu University of Traditional Chinese Medicine. 독성 효과가 있는 의약품에 대한 논의가 진행되던 회사로서, 독성 감쇠를 위한 Tiebangchui는 임상 적용에 매우 중요합니다. 고원 보리 와인을 사용한 Tiebangchui의 최적화 진행 기술은 간단하고 눈에 띄며 안정적입니다. 이는 산업 생산에 이러한 분산을 제공할 수 있습니다.엔트로피 방법과 결합된 Box-Behnken 설계는 최적화된 조건의 테이블 간 관찰을 가능하게 하며 화학, 약?...