Study on the Ethnic Medicine Column

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Editor: Invited Column Editor-in-chief： MA Shuang-cheng

Ethnic medicine is an important part of Chinese traditional medicine， and has formed its own unique medical theoretical system. In the process of thousands of years of mutual integration， most of the ethnic groups have formed the development status of "pluralism， integration and difference". The population of ethnic minorities in China accounts for 6% of the total population， but their settlements account for 60% of the total area of the country. There are more than 8 000 kinds of ethnic medicinal materials in China， accounting for more than 70% of the total medicinal materials resources. They belong to the cultural system of more than 40 different nationalities， and have biological diversity and cultural diversity. There are more than 600 varieties of ethnic medicine， 47 of which were listed in the national basic medical insurance drug catalogue by the national labor and social security department in 2000.

In 2012， the Division of Chinese Materia Medica of National Institutes for Food and Drug Control (hereinafter referred to as "NIFDC") established the department of the ethnic medicine and set up a technical platform for the ethnic medicine in the national drug-control system which aimed to explore a new model suitable for the development of quality control of national drugs， and improve the quality inspection level of national drugs. This reflects the great importance attached to the quality research of ethnic drugs， and is of milestone significance to the development of ethnic drug testing. From 2013 to 2014， the NIFDC planned and organized a research project on ethnic medicine， which was supported by special funds from the Drug Registration Administration Department of the National Medicine Products Administration (hereinafter referred to as the "NMPA"). Since 2015， the NIFDC， as the leading unit， has jointly carried out research work with 13 national provincial (district) drug control institutes. With the support of the NMPA， three phases of project research have been carried out， the draft quality standards for 25 kinds of characteristic ethnic medicinal materials have been completed， and the research results have formed a compilation of conclusion reports of more than 1 000 000 words. Since June 2021， the project has added five strong provincial drug inspection units to carry out cooperative research， and carried out research on 32 ethnic drug varieties.

In order to strengthen the research of ethnic medicine， "Research for the Ethnic Medicine Column"，including one comprehensive article and seven experimental article， has been specially set up in this issue. The paper—"Development and thinking of ethnic medicines based on patent analysis from 1994 to 2021" has analyzed the development status of ethnic medicine in China on the perspective of patent from 1994 to 2021. The results show that the ethnic medicine patent applications have experienced the embryonic stage from 1994 to 2003， the rapid development stage from 2003 to 2018 and the steady development stage from 2019 to now. It reflects that with the attention and support of the state to ethnic medicine and the efforts of scientific research institutions and scholars， some ethnic medicine work has made certain achievements. In the paper， "Identification of Polygoni Multiflori Radix， Dipsaci Radix， Morindae Officinalis Radix and Atractylodis Macrocephalae Rhizoma in Zishen Yutai pills by solid-phase extraction combined with thin layer chromatography"， the standard improvement research was carried out on Zishen Yutai pills which containing 15 herbs contained in the drug standard of the Ministry of Health of the People’s Republic of China， so as to lay a foundation for the follow-up study of ethnic medicine. In the paper， "Visualization analysis of spatial distribution of chemical compositions in Morindae Officinalis Radix processed product by matrix-assisted laser desorption ionization mass spectrometry imaging"， the matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) has used to acquire the data of spatial distribution of iridoid and saccharous in Morindae Officinalis Radix and the contents of iridoid in the samples were determined by UPLC which provides a new way of thinking for the Chinese medicine processing. In the paper， "Research on improvement of quality standard of the Guangxi ethnic herb Onychii Herba"， the quality standard and quality control level of Onychium japonicum (Thunb.) Kze. was improved. In the other four articles， "Comparison and optimization extraction of genomic DNA from Tibetan patent medicine Shiliu Jianwei powder"， "Based on specific primer for the HRM identification method study of Tibetan patent medicine Shiliu Jianwei powder". "Genomic DNA extraction conditions optimization of Arnebiae Radix based on response surface methodology" and "RFLP-HRM identification method of Arnebiae Radix based on DNA barcoding and high resolution melting"， DNA identification technology was used to optimize the extraction of genomic DNA from Arnebiae Radix and Shiliu Jianwei powder， so as to provide data support for the root cause of ethnic medicine.

Through the above research column， this issue hopes to provide readers with new ideas and new methods for the research of ethnic medicine， and expand the depth and breadth of ethnic medicine research， so as to promote the innovative development of ethnic medicine. And then to explore appropriate new models of ethnic medicine research， and jointly promote the development of basic research in the field of ethnic medicine.

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Chinese Journal of Pharmaceutical Analysis (8)

Publication year

2022 (8)

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Study on the Ethnic Medicine Column (8)

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Study on the Ethnic Medicine Column
Development and thinking of ethnic medicines based on patent analysis from 1994 to 2021^*

DAI Sheng-yun, LIU Jie, QIAO Fei, LIAN Chao-jie, GUO Li-nong, ZHENG Jian, MA Shuang-cheng

Chinese Journal of Pharmaceutical Analysis. 2022, 42(8): 1290-1305. https://doi.org/10.16155/j.0254-1793.2022.08.01

Abstract (15) PDF (4) Knowledge map Save

Taking “ethnic medicine” as the search term, this paper searches the patent information collected in CNKI patent database from 1994 to 2021, and analyzes the development status of ethnic medicine in China from the perspective of patent. Through the analysis of patent regional distribution, patent applicant types, patent technology fields, medicinal materials, prescription preparations and drug combinations involved in different ethnic medicine patents, the results show that the ethnic medicine patent applications have experienced the embryonic stage from 1994 to 2003, the rapid development stage from 2003 to 2018 and the steady development stage from 2019 to now. It reflects that with the attention and support of the state to ethnic medicine and the efforts of scientific research institutions and scholars, some ethnic medicine work has made certain achievements. The patent application areas, patent applicants and research institutions of ethnic medicines are regional, and the research emphases of different ethnic medicines are different. Some ethnic medicines have achieved the all-round development of industry, university and research, but there are still a large number of ethnic medicines waiting to be developed, and the quality standards of ethnic medicines have been improved. The key problems to be solved in the development of ethnic medicine are the rectification of the origin of ethnic medicine and the protection and sustainable utilization of ethnic medicinal resources.
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Study on the Ethnic Medicine Column
Identification of Polygoni Multiflori Radix, Dipsaci Radix, Morindae Officinalis Radix and Atractylodis Macrocephalae Rhizoma in Zishen Yutai pills by solid-phase extraction combined with thin layer chromatography

TANG Zhe, DAI Sheng-yun, WANG Wei, NING Na, LEI Ting, HE Zi-xin, QIAO Fei, ZHENG Jian, MA Shuang-cheng

Chinese Journal of Pharmaceutical Analysis. 2022, 42(8): 1306-1311. https://doi.org/10.16155/j.0254-1793.2022.08.02

Abstract (10) PDF (1) Knowledge map Save

Objective: To establish a thinlayer chromatographic method which combine with solid-phase extraction for the identification of Polygoni Multiflori Radix, Dipsaci Radix, Morindae Officinalis Radix and Atractylodis Macrocephalae Rhizoma in Zishen Yutai pills. Methods: C₁₈ solid phase extraction was used. The sample was loaded into the activated solidphase column after ultrasonic extraction, steam drying and redissolution of methanol. It was eluted with water and methanol of different concentrations successively, and the elution rate was 1 mL·min^-1. The water washing was collected, evaporated the filtrate to dryness and dissolved the residue in 1 mL of methanol to produce the test solution of Morindae Officinalis Radix, a mixture of chloroform, methanol, water and glacial acetic acid (7:3:1:1) was used as the mobile phase; 60% methanol eluent was collected, evaporated the filtrate to dryness and dissolved the residue in 1ml of methanol to produce the test solution of Dipsaci Radix identification, a mixture of upper layer solution of n-butanol, acetic acid and water (4:1:5) was used as the mobile phase; 80% methanol eluent was collected, evaporated the filtrate to dryness and dissolved the residue in 1 mL of methanol to produce the test solution of Polygoni Multiflori Radix, a mixture of Cyclohexane, dichloromethane, ethyl acetate, formic acid (10:1:1:0.1) was used as the mobile phase; 100% methanol eluent was collected, evaporated the filtrate to dryness and dissolved the residue in 1 mL of methanol to produce the test solution of Atractylodis Macrocephalae Rhizoma, n-hexane, ethyl acetate, formic acid (40:10:0.5) was used as the mobile phase. Results: The fluorescent spots in the chromatogram obtained with the test solution correspond in position and colour to the spots in the chromatogram obtained with the reference drug solution and reference solution, negative samples without interference. Conclusion: Solid phase extraction can effectively enrich and analyze the low-specific components in Zishen Yutai pills. The established method has the characteristics of simple operation and good repeatability, and can be used as thin layer chromatography identification of ZishenYutai pills.
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Study on the Ethnic Medicine Column
Visualization analysis of spatial distribution of chemical compositions in Morindae Officinalis Radix processed product by matrix-assisted laser desorption ionization mass spectrometry imaging

QIAO Fei, DAI Sheng-yun, LIAN Chao-jie, LIU Jie, DONG Jing, ZHENG Jian, MA Shuang-cheng

Chinese Journal of Pharmaceutical Analysis. 2022, 42(8): 1312-1318. https://doi.org/10.16155/j.0254-1793.2022.08.03

Abstract (10) PDF (2) Knowledge map Save

Objective: To visual analyze the changes of iridoid and saccharous in Morindae Officinalis Radix processed product by matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI). Methods: The data of spatial distribution of iridoid and saccharous in Morindae Officinalis Radix during different processing were acquired by MALDI-MSI. The contents of iridoid in the samples were determined by UPLC. The accuracy of MS imaging microscope was verified by the results of content determination. Results: The MS imaging microscope could distinguish different processed samples and realize the visualization analysis of spatial distribution of iridoid and saccharous in Morindae Officinalis Radix. The results were basically consistent with those determined by UPLC. Conclusion: MALDI-MSI can be used to study the processing quality of Morindae Officinalis Radix and this method provides new ideas for the Chinese medicine processing.
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Study on the Ethnic Medicine Column
Research on improvement of quality standard of the Guangxi ethnic herb Onychii Herba^*

LIN Que-yue, DAI Sheng-yun, XIE Pei-de, TENG Ai-jun, MA Shuang-cheng, LI Li-li, ZHENG Jian

Chinese Journal of Pharmaceutical Analysis. 2022, 42(8): 1319-1327. https://doi.org/10.16155/j.0254-1793.2022.08.04

Abstract (8) PDF (5) Knowledge map Save

Objective: To improve the quality standard of Onychii Herba and improve its quality control level. Methods: A resource survey and field collection of Onychii Herba were carried out in Guangxi. Microscopical characteristics of 17 batches of Onychii Herba and 5 batchs of Sphenomeridis Herba were studied based on transverse section and powder. TLC was adopted to distinguish between Onychii Herba and Sphenomeridis Herba. The water content, total ash, acid-insoluble ash and ethanol soluble extractives were determined according to the method in the appendix of Chinese Pharmacopoeia. The content of cichoric acid was determined by HPLC. The CAPCELLPAK MGⅡC₁₈ column(250 mm×4.6 mm, 5 μm) was used, the mobile phase was acetonitrile -0.3% phosphoric acid(19:81) at a flow rate of 1.0 mL·min^-1, the column temperature was 25 ℃, and the detection wavelength was at 330 nm. Results: Compared with Quality Standards for Zhuang Medicine in Guangxi Zhuang Autonomous Region, microscopically characteristics of transverse section was proposed, the improved TLC showed the spots developed clearly. The linear range of cichoric acid was 2.73-273 μg·mL^-1(r=1.000) and the average recovery was 102.8%(RSD=1.1%, n=6), the content of cichoric acid in 17 batches of samples ranged from 0.12% to 2.23%. Conclusion: The established method in this paper such as microscopic identification,TLC identification, inspection and content determination, improves the quality standardand provides evidence for further improvement of quality standards of Onychii Herba.
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Study on the Ethnic Medicine Column
Comparison and optimization extraction of genomic DNA from Tibetan patent medicine Shiliu Jianwei powder^*

FANG Wen-liang, LIU Jie, TANG Zhe, DAI Sheng-yun, LIAN Chao-jie, QIAO Fei, GUO Li-nong, MA Shuang-cheng, ZHENG Jian

Chinese Journal of Pharmaceutical Analysis. 2022, 42(8): 1328-1334. https://doi.org/10.16155/j.0254-1793.2022.08.05

Abstract (7) PDF (1) Knowledge map Save

Objective: To compare the effects of different kits and different methods on the extraction of genomic DNA from Tibetan patent medicine Shiliu Jianwei powder. Methods: The genomic DNA of the Tibetan patent medicine Shiliu Jianwei powder was extracted using four different commercially available kits and magnetic bead method and the adsorption column method. The effect of genomic DNA extraction was comprehensively compared with Nano Drop one, Qubit, and gel electrophoresis. Results: Four commercially available kits and two methods were used to extract the genomic DNA of the Shiliu Jianwei powder. The results showed that the adsorption column method, namely kit 3 and kit 4, was superior than the bead method using kit 1 and kit 2 in terms of the concentration of genomic DNA, the integrity of DNA fragments, and the PCR amplification effect, and economy. Conclusion: Preliminary research on the extraction effect of main stream nucleic acid extraction methods on genomic DNA of Tibetan patent medicine Shiliu Jianwei powder is performed. It is proved that under the standard method of the kit, the adsorption column method has a better extraction effect of genomic DNA, which lays a preliminary foundation for the further optimization of the extraction method.
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Study on the Ethnic Medicine Column
Based on specific primer for the HRM identification method study of Tibetan patent medicine Shiliu Jianwei powder^*

LIU Jie, FANG Wen-liang, TANG Zhe, QIAO Fei, LIAN Chao-jie, DAI Sheng-yun, GUO Li-nong, ZHENG Jian, MA Shuang-cheng

Chinese Journal of Pharmaceutical Analysis. 2022, 42(8): 1335-1344. https://doi.org/10.16155/j.0254-1793.2022.08.06

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Objective: To establish the high-resolution melting(HRM) identification method for different ingredients in Tibetan patent medicine Shiliu Jianwei powder. Methods: The internal transcribed spacer Ⅱ of nuclear ribosomal DNA(ITS2) sequence from the same or similar species of the active pharmaceutical ingredient (API) which was used as the template to design primers with Primer Primer 6.0 software, according to the primer design principle. The method of agarose gel electrophoresis was used to select the specific primers that showed better amplification effect on target APIs. The HRM method was used to verify the specific primers selected by agarose gel electrophoresis, and the consistency of the detection results of the homemade Shiliu Jianwei powder and the commercially available Shiliu Jianwei powder were compared. The number of amplification cycles of the HRM experiment was optimized, the best specific primers for each API was determined, and a more specific specific HRM method was established. Results: Fifty-two pairs of specific primers were designed in the experiment, and 11 pairs of specific primers with better amplification specificity were selected by agarose gel electrophoresis detection. The HRM method was used to verify the 11 pairs of specific primers selected by agarose gel electrophoresis, the specificity of the results was consistent with the results of agarose gel electrophoresis, and the test results in the homemade Shiliu Jianwei powder and the commercially available Shiliu Jianwei powder were consistent. The best specific primers in the 5 APIs were determined. The best specific primer for Semen Punicae was SLZ-7A7S (32 cycles), the best specific primer for Carthami Flos was HH-2A2S (25 cycles), the best specific primer for Cinnamomi Cortex was RGCA-1A1S (40 cycles), the best specific primer for Amomi Fructus Rotundus was DKAC-1A1S (27 cycles), and the best specific primer for Fructus Piperis Longi was BBPBN-1A1S (35 cycles). Conclusion: Finally, the optimal amplification primers and the optimal amplification cycle number of each API in the Tibetan patent medicine Shiliu Jianwei powder for HRM detection method are determined, and the more specific identification of each API in Shiliu Jianwei powder is realized. It provides research ideas and reference for the molecular biology quality control of Chinese patent medicines with original powder.
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Study on the Ethnic Medicine Column
Genomic DNA extraction condition optimization of Arnebiae Radix based on response surface methodology

LIU Jie, FANG Wen-liang, TANG Zhe, LIAN Chao-jie, DAI Sheng-yun, GUO Li-nong, ZHENG Jian, QIAO Fei, MA Shuang-cheng, LI Yun-zheng

Chinese Journal of Pharmaceutical Analysis. 2022, 42(8): 1345-1353. https://doi.org/10.16155/j.0254-1793.2022.08.07

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Objective: To optimize the genomic DNA extraction conditions of Arnebiae Radix based on response surface methodology. Methods: The effect of different operation steps' reaction time in the plant genomic DNA extraction kit on the extraction efficiency and quality of Arnebiae Radix genomic DNA was investigated. The 3 conditions sampling amount, ethanol treatment, and lysis time in the Arnebiae Radix extraction process were optimized based on response surface method. The optimized conditions were tested and verified. The PCR amplification system of Arnebiae Radix genomic DNA was optimized, by adjusting the primer concentrations in the PCR amplification system. Results: The investigation result of different operation steps' reaction time in the plant genomic DNA extraction kit on the extraction efficiency and quality of Arnebiae Radix genomic DNA, showed that there was no obvious effect of different reaction time. Based on the response surface methodology, the optimal extraction conditions for the genomic DNA of Arnebiae Radix were determined as sampling amount of 100 mg, lysing overnight (12-15 h), and soaking the samples in 100% ethanol for 24 h for pretreatment. Through the verification of 100 batches of Arnebiae Radix market samples, the success rate of PCR amplification was 89%. The optimal primer concentration in the PCR system of the Arnebiae Radix genomic DNA was determined to be 0.10 μmol·L^-1. If the genomic DNA still couldn't be amplified normally, the DNA can be diluted 10 times. Conclusion: The extraction and amplification efficiency of the Arnebiae Radix genomic DNA has been greatly improved, by optimizing the DNA extraction conditions and PCR amplification conditions. It provides support for the molecular biological identification for Arnebiae Radix. At the same time, it also provides a reference basis for the optimization of genomic DNA extraction conditions of other traditional Chinese medicine.
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Study on the Ethnic Medicine Column
RFLP-HRM identification method of Arnebiae Radix based on DNA barcoding and high resolution melting^*

LIU Jie, FANG Wen-liang, TANG Zhe, LIAN Chao-jie, DAI Sheng-yun, GUO Li-nong, ZHENG Jian, QIAO Fei, MA Shuang-cheng, LI Yun-zheng

Chinese Journal of Pharmaceutical Analysis. 2022, 42(8): 1354-1362. https://doi.org/10.16155/j.0254-1793.2022.08.08

Abstract (8) PDF (3) Knowledge map Save

Objective: To establish an identification method of restriction fragment length polymorphism-high resolution melting(RFLP-HRM) for Arnebiae Radix from medicine market based on DNA barcoding and high resolution melting(HRM). Methods: Internal transcribed spacer Ⅱ of nuclear ribosomal DNA (ITS2) sequences of Arnebiae Radix were amplified with polymerase chain reaction(PCR). MEGA 6.06 software was used to perform NJ tree cluster analysis and genetic distance analysis on the ITS2 sequence of Arnebiae Radix. Original plant confirmation of non-pharmacopoeial Arnebiae Radix from the market was performed. The primers were disigned using the software of Primer Premier 5 based on ITS2 sequence. The best one from 4 designed primers was screened through the HRM experiment. The identification method of RFLP-HRM for Arnebiae Radix was established. Results: The neighbour joining clustering analysis and genetic distance analysis of ITS2 sequences could effectively distinguish among the different species for this study. The HRM experiment confirmed Zcf-1 was the best primer. The RFLP-HRM method to effectively identify Arnebiae Radix from medicine market was established. Conclusion: The methods of DNA Barcoding and HRM both can effectively identify the Arnebiae Radix in this study, and the results are consistent and mutual authentication. The RFLP-HRM rapid identification method of Arnebiae Radix from medicine market has been established, which provides the reference basis for other traditional medicines.

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