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A study on the processing technology for Rhizoma Coptidis

A study on the processing technology for Rhizoma Coptidis Background: The present study intends to optimize the processing technology for the wine-processing of Rhizoma Coptidis, using alkaloids as indicators. Method: In the present study, the Box–Behnken design method was adopted to optimize the processing technol- ogy for Rhizoma Coptidis, using the alkaloid component quantities as the index. 100 g of Rhizoma Coptidis slices and 12.5 g of Rhizoma Coptidis wine were used. After full mixing, box-Behnken design method was used to optimize the processing time, processing temperature and processing time of coptis chinensis by taking alkaloid content as index. After mixing well, these components were fried in a container at 125 °C for 6 min and exhibited good parallelism. Results: The content of alkaloids in coptis chinensis was the highest after roasting at 125 °C for 6 min. The charac- teristic components were berberine hydrochloride, and the relative content was about 15.96%. And showed good parallelism. The effective components of Rhizoma Coptidis were primarily alkaloids. Conclusion: The optimized processing technology for Rhizoma Coptidis is good. Keywords: Wine coptis, Processing technology, Box–Behnken, Alkaloids, Yunlian Background heat to relieve heat toxicity in Traditional Chinese medi- Rhizoma Coptidis is the dry rhizome of Coptis chinensis, cine. It’s called catharsis and detoxification. This is the Coptis deltoidea, or Coptis teeta of the Ranunculaceae nature of traditional Chinese medicine (TCM) under the family [1–3], known as "Weilian", "Yalian" and "Yunlian," guidance of the theory of TCM. These Chinese medical respectively. Rhizoma Coptidis has a bitter and cold terms cannot be well translated into English, and there- nature and has the effect of clearing away heat, drying fore, pinyin is used. According to the theory of TCM, dampness, purging fire, and detoxifying [4, 5]. Purging bitter cold herbs usually have a thin Qi and a heavy taste fire and detoxifying is a term used in Traditional Chi - (coptis tastes bitter, gas cold, for the big bitter and big nese medicine (TCM). TCM believes that diseases can be cold goods, cannot be overserved, long served, raw good divided into cold syndrome and heat syndrome, requir- at clearing the heart fire. After processing, wine cop - ing symptomatic treatment. In Traditional Chinese medi- tis can mitigate the bitter cold in the raw food, lead up cine, diseases manifested by heat syndrome are usually medicine, good head of the fire). Qi and heavy taste are called heat toxicity, so there is a treatment of reducing also characteristics of Traditional Chinese medicine, and are classified under the guidance of traditional Chinese medicine theory. Taste has five tastes namely sour bitter *Correspondence: yangyongk291@163.com sweet salty. When processed with wine, it is possible to Department of Institute of Pharmacochemistry of Traditional Chinese both make use of the pungent hot properties and delivery Medicine, Chongqing Academy of Chinese Materia Medica, No. 34 of Nanshan Street, Nanan District, Chongqing 400065, China function of the wine to alleviate the bitter cold nature of Full list of author information is available at the end of the article © The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http:// creat iveco mmons. org/ licen ses/ by/4. 0/. The Creative Commons Public Domain Dedication waiver (http:// creat iveco mmons. org/ publi cdoma in/ zero/1. 0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Wang et al. BMC Biotechnology (2022) 22:3 Page 2 of 7 this herb, stop it from hurting the spleen and stomach, was 30  °C. In addition, the loading volume was 10 μL, and allow it to be cold without stagnating. The combina - and the herb underwent gradient elution with acetoni- tion of the hot and the cold allows it to play a better role trile-30  mmol/L ammonium bicarbonate (containing in clearing heat and purging fire [6, 7]. 0.7% ammonia and 0.1% triethylamine) as the mobile Usually, Chinese medicinal materials are the original phase, with a flow rate of 1  mL/min. The specific chro - plants, animals and minerals, which can not be directly matogram is shown in Figs. 1, and 3 shows the chromato- used for the treatment of diseases. They need to be pro - gram of the reference substances (Figs. 2, 3). cessed before they can be used, which is called yinpian, and the processing method and process is called process- The preparation of the test solution ing. The commonly used processing methods are steam - The Chinese medicine coptis chinensis is purchased ing, boiling and stir-frying. Sometimes it is necessary to from Chongqing Shizhu,Origin: Coptis chinensis Franch. add different ingredients for processing, such as wine, Dry rhizomes. A powder passing a 10-mesh sieve of the wheat bran, ginger, etc. Processing has the effect of alle - herb to be tested was obtained, and 0.2 g was placed in a viating and changing the curative effect. In this study, corked, conical flask, and precisely added with 50  ml of Coptis chinensis belongs to cold medicine in Traditional 50% acetonitrile-hydrochloric acid (100:1) mixed solu- Chinese medicine, and the wine used in processing is tion was added. The conical flask was tightly plugged, hot, which is processed to ease its bitter cold. In clinical weighed, treated with ultrasound (power 250  W, fre- practice, there are various methods for processing Rhi- quency 40 kHz) for 30 min, left standing for cooling, and zoma Coptidis with wine, e.g., adding rice wine (12.5 g of weighed. A 50% acetonitrile-hydrochloric acid (100:1) rice wine for every 100  g of Rhizoma Coptidis); mixing solution was added to make up the weight loss, and it was well; after the rice wine has been absorbed, placing it in shaken and filtered using qualitative filter paper. Of the a container preheated to 125  °C and frying it for 6  min; resultant filtrate, 2  mL was placed in a 10  mL measur - taking it out; and allowing it to cool. However, there is ing flask and a 50% acetonitrile-hydrochloric acid (100:1) no consensus over the best processing method [8–12]. solution was added until it reached the required weight. Therefore, the present study intends to optimize the pro - The test solution should be pretreated and filtered with cessing technology for the wine-processing of Rhizoma a 0.22 µm membrane before injection into HPLC. It was Coptidis, using alkaloids as indicators. The aim is to sug - shaken and filtered using a 0.22  μm microporous fil - gest a method for obtaining great quantities of the com- ter membrane. The resultant filtrate was the required ponents with a high degree of purity. solution. Methods Results Chromatographic conditions Single‑factor investigation of the processing technology The chromatographic conditions used in the present for the wine‑processing of Rhizoma Coptidis study were as follows: The column used was the Welch Investigation into the liquid absorption rate Xtimate C18 (4.6 × 250.0  mm, 3  μm), the detection Modern studies show that the following six alkaloids are wavelength was 270  nm, and the column temperature mainly found in coptis chinensis, which are: Medicine Fig. 1 High-performance liquid chromatogram. The numbered peaks represent: 2. Pharmacorhizine hydrochloride, 3. Tetrandrine hydrochloride, 4. Epiberberine hydrochloride, 5. Coptidine hydrochloride, 6. Palmatine hydrochloride, and 7. Berberine hydrochloride W ang et al. BMC Biotechnology (2022) 22:3 Page 3 of 7 Fig. 2 Contour and response surface Wang et al. BMC Biotechnology (2022) 22:3 Page 4 of 7 Fig. 3 Chromatogram of reference substances: 1. Epaperberine, 2. Coptidine, 3. Palmatine, and 4. Berberine Table 1 Alcohol absorption capacity of Rhizoma coptidis of Rhizoma Coptidis. Therefore, in the present study, 100 g of Rhizoma Coptidis and 12.5 g of yellow wine were Weighing sample (g) Add fluid volume (ml) Residual weighed and used. They were mixed well, and the absorp - fluid volume (ml) tion time was measured. The results revealed that the yel - low wine can be completely absorbed in 3 min. 100.00 500 400 100.00 405 100.00 405 Investigation into the processing time In the present study, 100 g of Rhizoma Coptidis and 12.5 g of yellow wine were weighed, mixed well, and stirred and stephania root alkali, hydrochloric acid Africa hydro- fried at 80 °C and 150 °C. The elapsed time was recorded. chloric acid alkali, hydrochloric acid table berberine and The results showed that the herb had a well-processed hydrastine hydrochloric acid, hydrochloric acid, Martin, appearance at 12 and 1.5 min, respectively. berberine hydrochloride (corresponding to peak in the chromatograph chart  2 ~ 7), but because of peak separa- tion effect is good, and the area is larger, thus will peak Investigation into the type of yellow wine and the alcohol respectively reference 1 ~ 7 as processing technology content research, but because of these seven elements only ber- In the present study, 12.5  g of different types of yellow berine hydrochloride in standard material, Therefore, the wine with varying alcohol contents were used. Each type chromatographic peak area of berberine hydrochloride of wine was mixed well with 100  g of Rhizoma Coptidis was used as reference. and fried at 115  °C for 7  min. The alkaloid content in In the present study, 100  g of Rhizoma Coptidis was each sample was then measured (see Table 2). The results weighed, 500 ml of yellow wine (manufacturer: Chengdu revealed that the alkaloid content in the wine-processing Julong Biological Technology Co., LTD., raw materials of Rhizoma Coptidis processed using different types of and accessories: water, rice, caramel color; The alcohol wine with varying alcohol concentrations was similar. is 10.0%vol; Total sugar: 15.0 g/L or less) was added, and the mixture was left to stand for one hour. The liquid was extracted, and the volume was measured (see Table  1). The processing technology for the wine‑processing The results showed that the absorption rate of Rhizoma of Rhizoma Coptidis designed by Box–Behnken Coptidis was approximately 1 ml/g. Factor‑level design With the processing temperature, processing time, and Investigation into the liquid absorption time moisture time as independent variables, seven character- According to the 2015 edition of the Chinese Pharma- istic peaks were measured to calculate the content per- copoeia, the procedures for the wine-processing of Rhi- centages, using berberine hydrochloride as a reference. zoma Coptidis are as follows: 100 g of Rhizoma Coptidis The sum of these seven characteristic peaks (Y) was used and 12.5  g of yellow wine (approximately 12.5  ml), and as the response value for the design. The experimental the amount of yellow wine should be far less than that design and results are shown in Tables 3 and 4. W ang et al. BMC Biotechnology (2022) 22:3 Page 5 of 7 Table 2 Alkaloid contents in different types of yellow rice wine and prepared alcoholic products (% by dry weight) Variety and alcohol of yellow rice Epiberberine (%) Coptisine (%) Palmatine (%) Berberine (%) wine Dry type,16% 1.33 2.26 2.23 7.95 Semi-dry,15% 1.29 2.22 2.21 7.98 Semi-Swee,10% 1.36 2.28 2.22 7.89 Sweet,11.5% 1.34 2.29 2.23 8.01 RSD% 2.22 1.37 0.44 0.65 2 2 2 Table 3 Ranges of design factors of Box-behnken B − 0.2837C, r = 0.8450. These results reveal that the goodness of fit of the model is good, and it can be used Level Moisture time/ Processing Processing for the analysis and prediction of the processing technol- min A temperature/°C B time/min C ogy. The processing temperature and processing time of Low 0 80 1.5 B and C factors had no significant effect on the content High 3 150 12 of alkaloids. The analysis of variance revealed that within the test range, the influencing factors for the processing technology were as follows (in decreasing order of the magnitude of influence): moisture time, processing time, Model fit and variance analysis and processing temperature (see Table 5). The content of all processed products showed different degrees of decrease when compared with the original medicinal materials. However, since the active ingredi- Optimization and prediction of the response surface ents were alkaloids, the total content (%) was chosen as The Design-Expert 11 software was used to draw the the evaluation index, with the aim of optimizing its maxi- response surface and a two-dimensional contour map mum value. The obtained regression model equation is of the moisture time (min), processing temperature (°C), as follows: Y = 15.23 − 0.3026A + 0.0333B + 0.2334C − and processing time (min) of the wine-processing of Rhi- 0.2121AB + 0.1991AC − 0.4357B C + 0.3969A − 0.5237 zoma Coptidis. Table 4 Box-behnken design test and results No A / min B/°C C/min Y1/% Y2/% Y3/% Y4/% Y5/% Y6/% Y7/% Y 1 3 150 6.75 0.18 0.39 0.58 1.41 2.28 2.13 7.77 2 0 80 6.75 0.16 0.40 0.61 1.38 2.33 2.17 7.99 3 1.5 80 1.5 0.15 0.39 0.52 1.26 2.14 1.95 7.03 4 3 115 12 0.16 0.40 0.64 1.41 2.29 2.25 7.93 5 1.5 80 12 0.17 0.43 0.65 1.35 2.36 2.23 8.01 6 1.5 115 6.75 0.16 0.41 0.65 1.35 2.26 2.21 7.94 7 1.5 115 6.75 0.20 0.42 0.66 1.49 2.31 2.22 7.83 8 0 150 6.75 0.18 0.42 0.64 1.46 2.35 2.29 8.07 9 3 115 1.5 0.17 0.41 0.64 1.38 2.19 2.23 7.61 10 1.5 115 6.75 0.18 0.41 0.67 1.46 2.29 2.27 8.04 11 1.5 150 12 0.17 0.40 0.64 1.33 2.14 2.20 7.65 12 1.5 150 1.5 0.18 0.42 0.62 1.39 2.21 2.19 7.52 13 3 80 6.75 0.19 0.42 0.64 1.43 2.36 2.21 7.98 14 1.5 115 6.75 0.18 0.41 0.67 1.46 2.35 2.28 8.15 15 0 115 12 0.18 0.42 0.68 1.47 2.35 2.38 8.16 16 1.5 115 6.75 0.18 0.42 0.69 1.43 2.32 2.24 7.93 17 0 115 1.5 0.19 0.42 0.67 1.45 2.51 2.33 8.42 Predictive value 0.178 0.420 0.683 1.500 2.440 2.356 8.376 15.964 Y1% to Y7% are equally important Wang et al. BMC Biotechnology (2022) 22:3 Page 6 of 7 Table 5 Variance analysis Source Sum of Squares df F‑ value P‑ value Model 4.36 9 0.4898 0.0350* Significant A-moisture time 0.7323 1 6.41 0.0392 B- processing temperature 0.0089 1 0.0775 0.7888 C- processing time 0.4358 1 3.81 0.0918 AB 0.1800 1 1.57 0.2498 AC 0.1585 1 1.39 0.2774 BC 0.7592 1 6.64 0.0366 A 0.6634 1 5.80 0.0468 B 1.15 1 10.10 0.0155 C 0.3388 1 2.96 0.1288 Residual 0.8001 7 Lack of Fit 0.6581 3 6.18 0.0555 Not significant Pure error 0.1420 4 Cor total 5.16 16 * indicates the significant value Discussion Table 6 Verify test results The TCM theory considers the following: the wine-pro - Y% (predicted value) Y% cessing of Rhizoma Coptidis can reduce its bitter-cold (measured nature and can lead the medicine upward, and mod- value) ern studies have revealed that the active ingredients 1 15.964 15.41 are alkaloids [13–15]. In addition, both processed and 2 15.38 unprocessed Rhizoma Coptidis have the same material 3 15.31 basis and similar alkaloid content but different effects. This needs to be investigated using pharmacodynamics [16–20]. In the present study, the Box–Behnken design method was adopted, and the alkaloid content was used With Y-max as the goal, the processing technology as the index to optimize the processing technology for was optimized, and the results were as follows: A = 0, the wine-processing of Rhizoma Coptidis. The results B = 125.792, and C = 5.828. Considering the actual revealed that the main active ingredients in the wine- situation of the instruments and equipment, the best processing of Rhizoma Coptidis are alkaloids. process was adjusted to the following: A = 0, B = 125, Limitations: The following limitations exist in the pre - and C = 6. This means that 100 g of Rhizoma Coptidis sent study. First, the pharmacological effect of the Rhi - and 12.5  g of yellow wine should be weighed, mixed zoma Coptidis after wine roasting is not clear in the well, and stirred and fried at 125 °C for 6 min. present study. Since the effective components of Rhizoma Using the optimal conditions, the value of Y1–Y7% Coptidis are alkaloids, in the present study, the process- and the value of Y are predicted as: Y1/% = 0.178, Y2% ing technology for Rhizoma Coptidis was optimized = 0.420,Y3% = 0.683,Y4% = 1.500,Y5 = 2.440,Y6 = 2.35 using alkaloids as indicators. However, according to the 6,Y7 = 8.376,Y = 15.964. theory of TCM, wine-processing can reduce the bitter cold nature of Rhizoma Coptidis and change its pharma- cological effect. This team plans to conduct further stud - Verification test ies on the pharmacological effects of the wine-processing In the present study, according to the proposed pro- of Rhizoma Coptidis. In addition, the clinical efficacy of cess, three batches of the processed product were pre- Rhizoma Coptidis after processing still needs to be fur- pared in parallel. The results revealed that the values ther verified through clinical research. of Y (%) for the three batches of the processed prod- Compared with traditional processing, the process- ucts were 15.41, 15.38, and 15.31 (see Table  6). The ing process was datalized and the processing end point data in Table 6 reveal that the optimized process in the was more objective. The traditional culture of processing present study exhibits good parallelism. W ang et al. BMC Biotechnology (2022) 22:3 Page 7 of 7 Received: 29 December 2020 Accepted: 20 December 2021 traditional Chinese medicine is mostly inherited by the master and apprentice, and the end point of processing is subjective judgment of human beings, which cannot form a unified standard. In this study, digital process param - References eters were used to specify the processing endpoint, which 1. Zhou S, Huang G, Chen G. Extraction, structural analysis, derivatization can combine traditional process with modern technology and antioxidant activity of polysaccharide from Chinese yam. Food Chem. 2021;361:130089. to a certain extent. 2. Finger JAFF, Lima EMF, Coelho KS, Behrens JH, Landgraf M, Franco BDGM, Pinto UM. Adherence to food hygiene and personal protection Conclusion recommendations for prevention of COVID-19. Trends Food Sci Technol. 2021;112:847–52. The main effective components of the wine-processing of 3. Chen F, Huang S, Huang G. Preparation, activity, and antioxidant mecha- Rhizoma Coptidis are alkaloids. The processing technol - nism of rice bran polysaccharide. Food Funct. 2021;12(2):834–9. ogy for the wine-processing of Rhizoma Coptidis is good 4. Chen F, Huang G, Huang H. 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Zhongguo WYH and QWH conceptualized and designed the study, drafted the initial Zhong yao za zhi Zhongguo zhongyao zazhi China J Chin Mater Med. manuscript, and reviewed and revised the manuscript.YYJ, BH, WJR, ZXM, GYL 1990;15(11):663–6. and HL designed the data collection instruments, collected data, carried out 9. Guo C. A sketch of the Coptis chinensis. Zhonghua Yi Shi Za Zhi. the initial analyses, and reviewed and revised the manuscript.YY coordinated 1985;15(4):196. and supervised data collection, and critically reviewed the manuscript for 10. Xu JT, Wang LQ, Xu B. Research development of Coptis chinensis. Zhong- important intellectual content. All authors approved the final manuscript as guo yi xue ke xue yuan xue bao. Acta Acad Med Sin. 2005;26(6):704–7. submitted and agree to be accountable for all aspects of the work. All authors 11. Zhang YL, Huang QW, Wang J, Su J, Li XF. Classification study of Coptis read and approved the final manuscript.. chinensis based on quantitative physical property characteristics of appearance and internal quality evaluation. China J Chin Mater Med. Funding 2012;37(20):3047–51. This study was supported by a grant from Project source: The Chongqing 12. Coptis chinensis franch. Evidence-based Complementary and Alternative Key Industry General Key Technology Innovation Project; Project name: The Medicine. 2012;2012(5):942384. research and demonstration of key technologies for the cultivation and 13. Wang DZ, Yi J, Zhang Y, Gao Q, Wang YZ, Li XG. [Study on optimal process- comprehensive development of genuine Chinese herbal medicines; Project ing technology of three kinds of processed coptidis rhizoma]. Zhong yao number: cstc2016zdcy-ztzx10005. The funding institutions only provided cai Zhongyaocai J Chin Med Mater. 2013;36(1):35–7. project demonstration, funding, and supervision of the research process, but 14. 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Optimization of microwave-assisted extraction of bioac- of the authors have any financial disclosure or conflict of interest. tive alkaloid compounds from Rhizoma Coptidis (Coptis chinensisFranch.). Food Sci Biotechnol. 2013;22(5):1–8. Author details Department of Institute of Pharmacochemistry of Traditional Chinese Medicine, Chongqing Academy of Chinese Materia Medica, No. 34 of Nanshan Publisher’s Note Street, Nanan District, Chongqing 400065, China. Chongqong Universit y Springer Nature remains neutral with regard to jurisdictional claims in pub- of Education, Chongqing 400067, China. lished maps and institutional affiliations. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png BMC Biotechnology Springer Journals

A study on the processing technology for Rhizoma Coptidis

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Abstract

Background: The present study intends to optimize the processing technology for the wine-processing of Rhizoma Coptidis, using alkaloids as indicators. Method: In the present study, the Box–Behnken design method was adopted to optimize the processing technol- ogy for Rhizoma Coptidis, using the alkaloid component quantities as the index. 100 g of Rhizoma Coptidis slices and 12.5 g of Rhizoma Coptidis wine were used. After full mixing, box-Behnken design method was used to optimize the processing time, processing temperature and processing time of coptis chinensis by taking alkaloid content as index. After mixing well, these components were fried in a container at 125 °C for 6 min and exhibited good parallelism. Results: The content of alkaloids in coptis chinensis was the highest after roasting at 125 °C for 6 min. The charac- teristic components were berberine hydrochloride, and the relative content was about 15.96%. And showed good parallelism. The effective components of Rhizoma Coptidis were primarily alkaloids. Conclusion: The optimized processing technology for Rhizoma Coptidis is good. Keywords: Wine coptis, Processing technology, Box–Behnken, Alkaloids, Yunlian Background heat to relieve heat toxicity in Traditional Chinese medi- Rhizoma Coptidis is the dry rhizome of Coptis chinensis, cine. It’s called catharsis and detoxification. This is the Coptis deltoidea, or Coptis teeta of the Ranunculaceae nature of traditional Chinese medicine (TCM) under the family [1–3], known as "Weilian", "Yalian" and "Yunlian," guidance of the theory of TCM. These Chinese medical respectively. Rhizoma Coptidis has a bitter and cold terms cannot be well translated into English, and there- nature and has the effect of clearing away heat, drying fore, pinyin is used. According to the theory of TCM, dampness, purging fire, and detoxifying [4, 5]. Purging bitter cold herbs usually have a thin Qi and a heavy taste fire and detoxifying is a term used in Traditional Chi - (coptis tastes bitter, gas cold, for the big bitter and big nese medicine (TCM). TCM believes that diseases can be cold goods, cannot be overserved, long served, raw good divided into cold syndrome and heat syndrome, requir- at clearing the heart fire. After processing, wine cop - ing symptomatic treatment. In Traditional Chinese medi- tis can mitigate the bitter cold in the raw food, lead up cine, diseases manifested by heat syndrome are usually medicine, good head of the fire). Qi and heavy taste are called heat toxicity, so there is a treatment of reducing also characteristics of Traditional Chinese medicine, and are classified under the guidance of traditional Chinese medicine theory. Taste has five tastes namely sour bitter *Correspondence: yangyongk291@163.com sweet salty. When processed with wine, it is possible to Department of Institute of Pharmacochemistry of Traditional Chinese both make use of the pungent hot properties and delivery Medicine, Chongqing Academy of Chinese Materia Medica, No. 34 of Nanshan Street, Nanan District, Chongqing 400065, China function of the wine to alleviate the bitter cold nature of Full list of author information is available at the end of the article © The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http:// creat iveco mmons. org/ licen ses/ by/4. 0/. The Creative Commons Public Domain Dedication waiver (http:// creat iveco mmons. org/ publi cdoma in/ zero/1. 0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Wang et al. BMC Biotechnology (2022) 22:3 Page 2 of 7 this herb, stop it from hurting the spleen and stomach, was 30  °C. In addition, the loading volume was 10 μL, and allow it to be cold without stagnating. The combina - and the herb underwent gradient elution with acetoni- tion of the hot and the cold allows it to play a better role trile-30  mmol/L ammonium bicarbonate (containing in clearing heat and purging fire [6, 7]. 0.7% ammonia and 0.1% triethylamine) as the mobile Usually, Chinese medicinal materials are the original phase, with a flow rate of 1  mL/min. The specific chro - plants, animals and minerals, which can not be directly matogram is shown in Figs. 1, and 3 shows the chromato- used for the treatment of diseases. They need to be pro - gram of the reference substances (Figs. 2, 3). cessed before they can be used, which is called yinpian, and the processing method and process is called process- The preparation of the test solution ing. The commonly used processing methods are steam - The Chinese medicine coptis chinensis is purchased ing, boiling and stir-frying. Sometimes it is necessary to from Chongqing Shizhu,Origin: Coptis chinensis Franch. add different ingredients for processing, such as wine, Dry rhizomes. A powder passing a 10-mesh sieve of the wheat bran, ginger, etc. Processing has the effect of alle - herb to be tested was obtained, and 0.2 g was placed in a viating and changing the curative effect. In this study, corked, conical flask, and precisely added with 50  ml of Coptis chinensis belongs to cold medicine in Traditional 50% acetonitrile-hydrochloric acid (100:1) mixed solu- Chinese medicine, and the wine used in processing is tion was added. The conical flask was tightly plugged, hot, which is processed to ease its bitter cold. In clinical weighed, treated with ultrasound (power 250  W, fre- practice, there are various methods for processing Rhi- quency 40 kHz) for 30 min, left standing for cooling, and zoma Coptidis with wine, e.g., adding rice wine (12.5 g of weighed. A 50% acetonitrile-hydrochloric acid (100:1) rice wine for every 100  g of Rhizoma Coptidis); mixing solution was added to make up the weight loss, and it was well; after the rice wine has been absorbed, placing it in shaken and filtered using qualitative filter paper. Of the a container preheated to 125  °C and frying it for 6  min; resultant filtrate, 2  mL was placed in a 10  mL measur - taking it out; and allowing it to cool. However, there is ing flask and a 50% acetonitrile-hydrochloric acid (100:1) no consensus over the best processing method [8–12]. solution was added until it reached the required weight. Therefore, the present study intends to optimize the pro - The test solution should be pretreated and filtered with cessing technology for the wine-processing of Rhizoma a 0.22 µm membrane before injection into HPLC. It was Coptidis, using alkaloids as indicators. The aim is to sug - shaken and filtered using a 0.22  μm microporous fil - gest a method for obtaining great quantities of the com- ter membrane. The resultant filtrate was the required ponents with a high degree of purity. solution. Methods Results Chromatographic conditions Single‑factor investigation of the processing technology The chromatographic conditions used in the present for the wine‑processing of Rhizoma Coptidis study were as follows: The column used was the Welch Investigation into the liquid absorption rate Xtimate C18 (4.6 × 250.0  mm, 3  μm), the detection Modern studies show that the following six alkaloids are wavelength was 270  nm, and the column temperature mainly found in coptis chinensis, which are: Medicine Fig. 1 High-performance liquid chromatogram. The numbered peaks represent: 2. Pharmacorhizine hydrochloride, 3. Tetrandrine hydrochloride, 4. Epiberberine hydrochloride, 5. Coptidine hydrochloride, 6. Palmatine hydrochloride, and 7. Berberine hydrochloride W ang et al. BMC Biotechnology (2022) 22:3 Page 3 of 7 Fig. 2 Contour and response surface Wang et al. BMC Biotechnology (2022) 22:3 Page 4 of 7 Fig. 3 Chromatogram of reference substances: 1. Epaperberine, 2. Coptidine, 3. Palmatine, and 4. Berberine Table 1 Alcohol absorption capacity of Rhizoma coptidis of Rhizoma Coptidis. Therefore, in the present study, 100 g of Rhizoma Coptidis and 12.5 g of yellow wine were Weighing sample (g) Add fluid volume (ml) Residual weighed and used. They were mixed well, and the absorp - fluid volume (ml) tion time was measured. The results revealed that the yel - low wine can be completely absorbed in 3 min. 100.00 500 400 100.00 405 100.00 405 Investigation into the processing time In the present study, 100 g of Rhizoma Coptidis and 12.5 g of yellow wine were weighed, mixed well, and stirred and stephania root alkali, hydrochloric acid Africa hydro- fried at 80 °C and 150 °C. The elapsed time was recorded. chloric acid alkali, hydrochloric acid table berberine and The results showed that the herb had a well-processed hydrastine hydrochloric acid, hydrochloric acid, Martin, appearance at 12 and 1.5 min, respectively. berberine hydrochloride (corresponding to peak in the chromatograph chart  2 ~ 7), but because of peak separa- tion effect is good, and the area is larger, thus will peak Investigation into the type of yellow wine and the alcohol respectively reference 1 ~ 7 as processing technology content research, but because of these seven elements only ber- In the present study, 12.5  g of different types of yellow berine hydrochloride in standard material, Therefore, the wine with varying alcohol contents were used. Each type chromatographic peak area of berberine hydrochloride of wine was mixed well with 100  g of Rhizoma Coptidis was used as reference. and fried at 115  °C for 7  min. The alkaloid content in In the present study, 100  g of Rhizoma Coptidis was each sample was then measured (see Table 2). The results weighed, 500 ml of yellow wine (manufacturer: Chengdu revealed that the alkaloid content in the wine-processing Julong Biological Technology Co., LTD., raw materials of Rhizoma Coptidis processed using different types of and accessories: water, rice, caramel color; The alcohol wine with varying alcohol concentrations was similar. is 10.0%vol; Total sugar: 15.0 g/L or less) was added, and the mixture was left to stand for one hour. The liquid was extracted, and the volume was measured (see Table  1). The processing technology for the wine‑processing The results showed that the absorption rate of Rhizoma of Rhizoma Coptidis designed by Box–Behnken Coptidis was approximately 1 ml/g. Factor‑level design With the processing temperature, processing time, and Investigation into the liquid absorption time moisture time as independent variables, seven character- According to the 2015 edition of the Chinese Pharma- istic peaks were measured to calculate the content per- copoeia, the procedures for the wine-processing of Rhi- centages, using berberine hydrochloride as a reference. zoma Coptidis are as follows: 100 g of Rhizoma Coptidis The sum of these seven characteristic peaks (Y) was used and 12.5  g of yellow wine (approximately 12.5  ml), and as the response value for the design. The experimental the amount of yellow wine should be far less than that design and results are shown in Tables 3 and 4. W ang et al. BMC Biotechnology (2022) 22:3 Page 5 of 7 Table 2 Alkaloid contents in different types of yellow rice wine and prepared alcoholic products (% by dry weight) Variety and alcohol of yellow rice Epiberberine (%) Coptisine (%) Palmatine (%) Berberine (%) wine Dry type,16% 1.33 2.26 2.23 7.95 Semi-dry,15% 1.29 2.22 2.21 7.98 Semi-Swee,10% 1.36 2.28 2.22 7.89 Sweet,11.5% 1.34 2.29 2.23 8.01 RSD% 2.22 1.37 0.44 0.65 2 2 2 Table 3 Ranges of design factors of Box-behnken B − 0.2837C, r = 0.8450. These results reveal that the goodness of fit of the model is good, and it can be used Level Moisture time/ Processing Processing for the analysis and prediction of the processing technol- min A temperature/°C B time/min C ogy. The processing temperature and processing time of Low 0 80 1.5 B and C factors had no significant effect on the content High 3 150 12 of alkaloids. The analysis of variance revealed that within the test range, the influencing factors for the processing technology were as follows (in decreasing order of the magnitude of influence): moisture time, processing time, Model fit and variance analysis and processing temperature (see Table 5). The content of all processed products showed different degrees of decrease when compared with the original medicinal materials. However, since the active ingredi- Optimization and prediction of the response surface ents were alkaloids, the total content (%) was chosen as The Design-Expert 11 software was used to draw the the evaluation index, with the aim of optimizing its maxi- response surface and a two-dimensional contour map mum value. The obtained regression model equation is of the moisture time (min), processing temperature (°C), as follows: Y = 15.23 − 0.3026A + 0.0333B + 0.2334C − and processing time (min) of the wine-processing of Rhi- 0.2121AB + 0.1991AC − 0.4357B C + 0.3969A − 0.5237 zoma Coptidis. Table 4 Box-behnken design test and results No A / min B/°C C/min Y1/% Y2/% Y3/% Y4/% Y5/% Y6/% Y7/% Y 1 3 150 6.75 0.18 0.39 0.58 1.41 2.28 2.13 7.77 2 0 80 6.75 0.16 0.40 0.61 1.38 2.33 2.17 7.99 3 1.5 80 1.5 0.15 0.39 0.52 1.26 2.14 1.95 7.03 4 3 115 12 0.16 0.40 0.64 1.41 2.29 2.25 7.93 5 1.5 80 12 0.17 0.43 0.65 1.35 2.36 2.23 8.01 6 1.5 115 6.75 0.16 0.41 0.65 1.35 2.26 2.21 7.94 7 1.5 115 6.75 0.20 0.42 0.66 1.49 2.31 2.22 7.83 8 0 150 6.75 0.18 0.42 0.64 1.46 2.35 2.29 8.07 9 3 115 1.5 0.17 0.41 0.64 1.38 2.19 2.23 7.61 10 1.5 115 6.75 0.18 0.41 0.67 1.46 2.29 2.27 8.04 11 1.5 150 12 0.17 0.40 0.64 1.33 2.14 2.20 7.65 12 1.5 150 1.5 0.18 0.42 0.62 1.39 2.21 2.19 7.52 13 3 80 6.75 0.19 0.42 0.64 1.43 2.36 2.21 7.98 14 1.5 115 6.75 0.18 0.41 0.67 1.46 2.35 2.28 8.15 15 0 115 12 0.18 0.42 0.68 1.47 2.35 2.38 8.16 16 1.5 115 6.75 0.18 0.42 0.69 1.43 2.32 2.24 7.93 17 0 115 1.5 0.19 0.42 0.67 1.45 2.51 2.33 8.42 Predictive value 0.178 0.420 0.683 1.500 2.440 2.356 8.376 15.964 Y1% to Y7% are equally important Wang et al. BMC Biotechnology (2022) 22:3 Page 6 of 7 Table 5 Variance analysis Source Sum of Squares df F‑ value P‑ value Model 4.36 9 0.4898 0.0350* Significant A-moisture time 0.7323 1 6.41 0.0392 B- processing temperature 0.0089 1 0.0775 0.7888 C- processing time 0.4358 1 3.81 0.0918 AB 0.1800 1 1.57 0.2498 AC 0.1585 1 1.39 0.2774 BC 0.7592 1 6.64 0.0366 A 0.6634 1 5.80 0.0468 B 1.15 1 10.10 0.0155 C 0.3388 1 2.96 0.1288 Residual 0.8001 7 Lack of Fit 0.6581 3 6.18 0.0555 Not significant Pure error 0.1420 4 Cor total 5.16 16 * indicates the significant value Discussion Table 6 Verify test results The TCM theory considers the following: the wine-pro - Y% (predicted value) Y% cessing of Rhizoma Coptidis can reduce its bitter-cold (measured nature and can lead the medicine upward, and mod- value) ern studies have revealed that the active ingredients 1 15.964 15.41 are alkaloids [13–15]. In addition, both processed and 2 15.38 unprocessed Rhizoma Coptidis have the same material 3 15.31 basis and similar alkaloid content but different effects. This needs to be investigated using pharmacodynamics [16–20]. In the present study, the Box–Behnken design method was adopted, and the alkaloid content was used With Y-max as the goal, the processing technology as the index to optimize the processing technology for was optimized, and the results were as follows: A = 0, the wine-processing of Rhizoma Coptidis. The results B = 125.792, and C = 5.828. Considering the actual revealed that the main active ingredients in the wine- situation of the instruments and equipment, the best processing of Rhizoma Coptidis are alkaloids. process was adjusted to the following: A = 0, B = 125, Limitations: The following limitations exist in the pre - and C = 6. This means that 100 g of Rhizoma Coptidis sent study. First, the pharmacological effect of the Rhi - and 12.5  g of yellow wine should be weighed, mixed zoma Coptidis after wine roasting is not clear in the well, and stirred and fried at 125 °C for 6 min. present study. Since the effective components of Rhizoma Using the optimal conditions, the value of Y1–Y7% Coptidis are alkaloids, in the present study, the process- and the value of Y are predicted as: Y1/% = 0.178, Y2% ing technology for Rhizoma Coptidis was optimized = 0.420,Y3% = 0.683,Y4% = 1.500,Y5 = 2.440,Y6 = 2.35 using alkaloids as indicators. However, according to the 6,Y7 = 8.376,Y = 15.964. theory of TCM, wine-processing can reduce the bitter cold nature of Rhizoma Coptidis and change its pharma- cological effect. This team plans to conduct further stud - Verification test ies on the pharmacological effects of the wine-processing In the present study, according to the proposed pro- of Rhizoma Coptidis. In addition, the clinical efficacy of cess, three batches of the processed product were pre- Rhizoma Coptidis after processing still needs to be fur- pared in parallel. The results revealed that the values ther verified through clinical research. of Y (%) for the three batches of the processed prod- Compared with traditional processing, the process- ucts were 15.41, 15.38, and 15.31 (see Table  6). The ing process was datalized and the processing end point data in Table 6 reveal that the optimized process in the was more objective. The traditional culture of processing present study exhibits good parallelism. W ang et al. BMC Biotechnology (2022) 22:3 Page 7 of 7 Received: 29 December 2020 Accepted: 20 December 2021 traditional Chinese medicine is mostly inherited by the master and apprentice, and the end point of processing is subjective judgment of human beings, which cannot form a unified standard. 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Optimization of microwave-assisted extraction of bioac- of the authors have any financial disclosure or conflict of interest. tive alkaloid compounds from Rhizoma Coptidis (Coptis chinensisFranch.). Food Sci Biotechnol. 2013;22(5):1–8. Author details Department of Institute of Pharmacochemistry of Traditional Chinese Medicine, Chongqing Academy of Chinese Materia Medica, No. 34 of Nanshan Publisher’s Note Street, Nanan District, Chongqing 400065, China. Chongqong Universit y Springer Nature remains neutral with regard to jurisdictional claims in pub- of Education, Chongqing 400067, China. lished maps and institutional affiliations.

Journal

BMC BiotechnologySpringer Journals

Published: Jan 14, 2022

Keywords: Wine coptis; Processing technology; Box–Behnken; Alkaloids; Yunlian

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