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

Summary

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.

Abstract

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 °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.

Introduction

The dried root of Aconitum pendulum Busch and A. flavum Hand.-Mazz., one of the most famous Tibetan medicines, is called Tiebangchui (TBC) in Chinese^1,2. 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 it....

Protocol

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

1. 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 electronic analytical balance and place them in a 10 mL volumetric flask. Then, add 0.05% hydrochloric acid methanol solution to dissolve the solids and ....

Results

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-.......

Discussion

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 years^24,25,26. Diterpenoid alkaloids are both active and toxic ingredients of TBC^27,28,29. The main toxic effects of the aconitum alkaloids of T.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
3-Deoxyaconitine	Chengdu Desite Biotechnology Co., Ltd.	DST221109-033
Aconitine	Chengdu Desite Biotechnology Co., Ltd.	DSTDW000602
Ammonium acetate	Tianjin Kermel Chemical Reagent Co., Ltd	Chromatographic grade
Benzoylaconitine	Chengdu Desite Biotechnology Co., Ltd.	DSTDB005502
Design-Expert software	Stat-Ease, Inc., Minneapolis, MN, USA	version 13.0
Electronic analytical balance	Shanghai Liangping Instruments Co., Ltd.	FA1004
High performance liquid chromatography	SHIMADZU Co., Ltd.	LC-20A
High-speed smashing machine	Beijing Zhongxing Weiye Instrument Co., Ltd.	FW-100
Millipore filter	Tianjin Jinteng Experimental Equipment Co., Ltd	φ13 0.22 Nylon66
stir-Fry machine	Changzhou Maisi Machinery Co., Ltd	Type 5
Tiebangchui	Gannan Baicao Biotechnology Development Co., Ltd	20211012
Ultra pure water systemic	RephiLe Bioscience, Ltd.	Genie G
Ultrasonic cleansing machine	Ningbo Xinyi Ultrasonic Equipment Co., Ltd	SB2200
Zanba	27 Chuanzang Road, Ganzi County	-