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Abstract ［Objectives］ This study was conducted to investigate the effects of seeding rate and light on the seed yield and active ingredients of Vernonia anthelmintica.
［Methods］With V. anthelmintica L. Willd as the research object, the seed yield was determined by staged harvesting combined with the actual cutting and measuring method; and the caffeoylquinic acids (CQA) were used as the characterization indexes, and high performance liquid chromatography was used to determine the contents of 3,4-CQA, 3,5-CQA and 4,5-CQA of V. anthelmintica seeds under different seeding rate+light treatments, so as to compare the changes in seed yield and chemical composition of V. anthelmintica under different seeding rates and light factors.
［Results］ In Moyu County, Hetian and similar areas, the suitable planting rate of V. anthelmintica was in the range of 13.5-15.0 kg/hm2. For different harvesting purposes, the optimal seeding rate and light conditions of V. Anthelmintica were different.
［Conclusions］The seeding rate and light conditions can be used to interfere with the seed yield and quality of V. anthelmintica.
Key words Vernonia anthelmintica; Seeding rate; Light; Seed quality; Chemical component; Mass fraction
Received: January 3, 2021 Accepted: March 4, 2021
Supported by Autonomous Region Science and Technology Supporting Xinjiang Project Plant Project (2016E02007); the West Light Foundation of The Chinese Academy of Sciences(2019-FPGGRC-004).
Yanrui CHEN (1976-), female, associate researcher, PhD, devoted to research about medicinal plant resources and basic medicine.
*Corresponding author. E-mail: [email protected].
Vernonia anthelmintica (L.) Willd, also known as Indian Shanhuixiang and Aiterilali, is an annual tall herb in Vernonia of Compositae. The stems are erect, thick, stronge, and the achenes are nearly cylindrical, dark brown or black, about 3-5 mm in length, and named Kalaziran and Kaliziri in Uyghur medicine［1-2］. The mature achenes of V. anthelmintica is an important medicinal material in traditional medicine, which has the functions of expelling parasite, subsiding swelling, and eliminating cold to stop pain, and is mainly used to treat vitiligo, damp-cold stomach pain, liver disease, inflammation and intestinal parasites［3］. The single medicine and compound preparations developed with V. anthelmintica as the main medicine include injections, ointments, liniments, tablets, pills, etc.［4-6］, among which Compound Quchongbanjiuju Pill and Quchongbanjiuju injection are widely used in clinical practice and have good effects on the treatment of vitiligo［7-8］. As a main medicinal material in the commonly used prescriptions for the treatment of vitiligo in Uyghur medicine, the main anti-vitiligo components isolated from V. anthelmintica are caffeoylquinic acids and sesquiterpenes［9-11］. Our research team established the fingerprint of V. anthelmintica based on HPTLC and HPLC-PDA technology, used HPLC-QTOF-MS to identify the main components of the medicinal material combinining with statistical analysis, and found that the main active ingredients of V. anthelmintica seeds are 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid［10］. According to literatures, V. anthelmintica is distributed in India, Pakistan, Sri Lanka, Nepal, Afghanistan, Myanmar, Laos, Malaysia and Chinas western Yunnan, Xinjiang (Hotan, Aksu, etc.), Guangxi Province, Shaanxi Province, etc.［12］. V. anthelmintica seeds are mainly imported, and there is currently no large-scale production and no normal seed sales channels. The seeds are dormant, and the germination rate of the purchased seeds is usually between 5% and 65%, which hinders the planting and seed production of V. anthelmintica. In order to keep the internal quality and clinical efficacy of V. anthelmintica decoction and various preparations from being affected by the quality of its seeds as a medicinal material, on the basis of the previous research, the effects of seeding rate and light on the seed yield and active ingredients of V. anthelmintica were investigated, aiming to improve the seed yield and quality of V. anthelmintica through the regulation of the seeding rate and light conditions and to further lay a foundation for V. anthelmintica planting and production and anti-vitiligo research.
Materials and Methods
Sources of test seeds
V. anthelmintica seeds were purchased from Kashgar Kunlun Pharmaceutical Co., Ltd. of Xinjiang in 2015. The original medicinal material was identified as the mature seeds of V. anthelmintica in Vernonia of Compositae by Associate Researcher Feng Ying of Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences. The specimen (WY02316) is deposited in the Key Laboratory of Plant Resources and Chemistry of Arid Zone, Chinese Academy of Sciences. We have carried out a standardized planting test of V. anthelmintica in Moyu County, Hotan, Xinjiang since 2015. The seeding material for this test was the self-propagated and self-retained seed material purchased above.
Design of field trials
The experimental area is located in Moyu Agricultural Science and Technology Park, Zawa Town, Moyu County, Hotan Prefecture, at E 79°37′, N 37°09′, and 1 386 m above sea level. The planting area was 1.5 hm2. The annual average temperature in this area is 11.4 ℃; and the accumulated temperature on the limit of above 0 ℃ is 4 450 ℃, and that on the limit equal to or greater than 10 ℃ is 4 017 ℃. The frost-free period is about 210 d. The main source of irrigation water was groundwater. The soil is fertile sandy loam with a pH value of 7.3-7.6. The 1 000-seed weight was 4.22 g, and the total annual irrigation amount was 9 000-12 000 m3/hm2. The experiment adopted an orthogonal experiment design, and set with 9 treatments, in 3 replicates, i.e., 27 plots in total. Seeding was carried out in early April with a planting depth of 1.0-2.0 cm and a row spacing of 40 cm. Based on the previous seeding rate control experiment, the seeding rate of V. anthelmintica was set to 12.0, 13.5 and 15.0 kg/hm2, and the light was divided into two layers of shading (Z2), one layer of shading (Z1), and no shading (CK), as shown in Table 1.
The phenology and plant growth changes of V. anthelmintica were observed every 10 d; during the seed maturiation period of V. anthelmintica, harvesting, drying, and sieving were carried out in stages according to the seed maturity conditions in plots.
Testing methods
Seed yield testing method
Seed yield was measured by the actual cutting method, which is to collect V. anthelmintica seeds and harvest from the harvested stems, then weigh them with scales, water meters, etc., and measure impurities.
Method for determination of active ingredient contents
The contents of 3,4-dicaffeoylquinic acid (3,4-CQA), 3,5-dicaffeoylquinic acid (3,5-CQA), and 4,5-dicaffeoylquinic acid (4,5-CQA) and the content of total caffeoylquinic acid (3 total CQA) in V. anthelmintica seeds were determined by HPLC.
Data processing methods
SPSS16.0 and Originpro9.0 were used for data statistics and analysis.
Results and Analysis
Effect of seeding rate+light on seed yield of V. anthelmintica
V. anthelmintica seeds were matured in stages, generally from mid-to-late August to early October. At first, mature V. anthelmintica seeds in each plot were harvested daily, dried, cleaned and stored separately; by the beginning of October, when the seeds on the plants were basically mature, V. anthelmintica stalks were harvested by plot at one time, and the seeds were collected, dried, cleaned and stored separately. Then, the yields of V. anthelmintica seeds under different treatment conditions were measured, and the relationship between each seeding rate+light treatment and the seed yield of V. anthelmintica is shown in Fig. 1 below.
From Fig. 1 above, it can be seen that there were differences in the seed yield of V. anthelmintica under various seeding rate+shading treatment conditions. The differences in the seed yield of V. anthelmintica under different seeding rate+shading treatment conditions were rather large. Treatment 8 (seeding rate 15.0 kg/hm2+one shading layer) had the highest yield of V. anthelmintica, followed by treatment 7 (seeding rate 15.0 kg/hm2+two shading layers), while treatment 9 (seeding rate 15.0 kg/hm2+no shading) showed the lowest seed yield. Effects of seeding rate+light on the active ingredients of V. anthelmintica
The contents of 3,4-CQA, 3,5-CQA, 4,5-CQA and total CQA of V. anthelmintica seeds under different seeding rates+light conditions were tested and analyzed. The results are shown in Fig. 2 below.
It can be seen from Fig. 2 that under different seeding rate+light treatment conditions, the contents of 3,4-CQA, 3,5-CQA, 4,5-CQA, and total CQA in V. anthelmintica seeds were different, and the contents of 3,5-CQA and total CQA content varied greatly, while the contents of 3,4-CQA and 4,5-CQA varied very little. Specifically, under the condition of treatment 8 (the seeding rate of 15 kg/hm2 and one layer of shading), the 4,5-CQA and total CQA contents of V. anthelmintica seeds were significantly higher than other treatments; treatment 6 (the seeding rate of 13.5 kg/hm2, no shading) showed a higher total CQA content, while the 3,5-CQA content was higher than other treatments; treatment 9 (the seeding rate of 15.0 kg/hm2+no shading) showed a 3,4-CQA content higher than other treatments; under the condition of treatment 2 (the seeding rate of 12.0 kg/hm2+one layer of shading), the contents of 3,4-CQA, 3,5-CQA and total CQA were lower than other treatments; and under the condition of treatment 1 (the seeding rate of 12.0 kg/hm2+two layers of shading), the content of 4,5-CQA was lower than other treatments. It can be seen that the seeding rate and light had important effects on the quality of V. anthelmintica.
Conclusions
After 3 years of seeding density control gradient test in the early stage, we concluded that the suitable seeding rate for V. anthelmintica in Moyu County, Hotan and similar areas was 12.0-15.0 kg/hm2. Based on this, the effects of different seeding rate+light conditions on the seed yield and active ingredients of V. anthelmintica were investigated in this study, and the results showed that the suitable seeding rate of V. anthelmintica was 13.5-15.0 kg/hm2.
① If the purpose is to harvest the seeds of V. anthelmintica, the suitable sowing rate in Moyu County, Hotan and areas with similar climatic conditions can be 15.0 kg/hm2, and proper shading during the vegetative growth period can increase its yield.
② If the purpose is to extract 4,5-CQA and total CQA in V. anthelmintica seeds, the optimal seeding rate and light for planting V. anthelmintica can be 15 kg/hm2 and one layer of shading. For the purpose of 3,5-CQA in V. anthelmintica seeds, the optimal seeding rate and light for planting V. anthelmintica recommended are 13.5 kg/hm2 and no shading. If 3,4-CQA is to be extracted from V. anthelmintica seeds, the optimal seeding rate and light can be 15.0 kg/hm2 and no shading. ③ The seed yield of V. anthelmintica and the contents of 3,4-CQA, 3,5-CQA and total CQA under the condition of 12.0 kg/hm2+one layer of shading were lower than other treatments, while under the condition of the seeding rate 12 kg/hm2+two layers of shading, the content of 4,5-CQA in seeds was lower than other treatments.
Agricultural Biotechnology2021
References
［1］ Chinese Pharmacopoeia Commission. Drug Standards of the Ministry of Health, the Peoples Republic of China-Uyghur Medicines: Uyghur Medicine Section［S］. Urumchi: Xinjiang Science and Technology Health Press, 1998. (in Chinese)
［2］ The state Administration of Traditional Chinese Medicine of Chinese Materia Medica editorial board. Chinese materia medica, Uygur part［M］. Shanghai: Shanghai science and Technology Press, 2005: 206-207. (in Chinese)
［3］ SHI DR, TIAN HY, LIU JL, et al. Preliminary study on the treatment of vitiligo with Uyghur medicine and western medicine［J］. Xinjiang Zhongyiyao, 2005,23(1): 31-32. (in Chinese)
［4］ National Institutes for Food and Drug Control. Records of Chinese ethnodrugs［M］. Beijing: Peoples Medical Publishing House. (in Chinese)
［5］ The Editorial Board of Flora of Xinjiang. Flora of Xinjiang［M］. Urumchi: Xinjiang Science and Technology Health Press, 1999. (in Chinese)
［6］ LIU YM, SAWET YIKIM. Records of Uyghur medicine［M］. Urumchi: Xinjiang Science and Technology Health Press,1993. (in Chinese)
［7］ HURXIGUL MAMAT. Clinical observation on 90 cases of vitiligo treated by Uygur medicine［J］. Jornal of Xlnnjiang Ulghur medical College, 1997(3): 26-31. (in Chinese)
［8］ TURSUN UPUR, ANIWAR JAMAL. Observation on curative effect of uigurian medicine in treating 610 cases suffered from Pityriasis versicolor alba［J］ . Journal of Medicine & Pharmacy of Chinese Minoritie, 2003(1): 12. (in Chinese)
［9］ LIU ZY, ZAN K, WU X, et al. Research progress on chemical constituents, quality control and pharmacological activity of Vernonia anthelmintica［J］. Chinese Journal of Pharmaceutical Analysis, 2020, 40(2): 199-208. (in Chinese)
［10］ MAIMAITI ZULIPIYA. Study on the optimization of the preparation process and quality standards of the active ingredients from Vernonia anthelmintica［D］. Beijing: University of Chinese Academy of Sciences, 2019. (in Chinese)
［11］ ABDUWAR ABDULHAK, SALAMET ELY, WANG YL. Studies on quality control of uyghur crude drug coriander seeds of Vernonia anthelmintica Willd［J］. Journal of Medicine & Pharmacy of Chinese Minoritie, 2015, 21(4): 44-46. (in Chinese)
［12］ L U TH. Research and application of uighur medicine Vernonia anthelmintica in the treatment of vitiligo［J］. Foreign Medical Sciences, 2004, 26(2): 103-104. (in Chinese)
［Methods］With V. anthelmintica L. Willd as the research object, the seed yield was determined by staged harvesting combined with the actual cutting and measuring method; and the caffeoylquinic acids (CQA) were used as the characterization indexes, and high performance liquid chromatography was used to determine the contents of 3,4-CQA, 3,5-CQA and 4,5-CQA of V. anthelmintica seeds under different seeding rate+light treatments, so as to compare the changes in seed yield and chemical composition of V. anthelmintica under different seeding rates and light factors.
［Results］ In Moyu County, Hetian and similar areas, the suitable planting rate of V. anthelmintica was in the range of 13.5-15.0 kg/hm2. For different harvesting purposes, the optimal seeding rate and light conditions of V. Anthelmintica were different.
［Conclusions］The seeding rate and light conditions can be used to interfere with the seed yield and quality of V. anthelmintica.
Key words Vernonia anthelmintica; Seeding rate; Light; Seed quality; Chemical component; Mass fraction
Received: January 3, 2021 Accepted: March 4, 2021
Supported by Autonomous Region Science and Technology Supporting Xinjiang Project Plant Project (2016E02007); the West Light Foundation of The Chinese Academy of Sciences(2019-FPGGRC-004).
Yanrui CHEN (1976-), female, associate researcher, PhD, devoted to research about medicinal plant resources and basic medicine.
*Corresponding author. E-mail: [email protected].
Vernonia anthelmintica (L.) Willd, also known as Indian Shanhuixiang and Aiterilali, is an annual tall herb in Vernonia of Compositae. The stems are erect, thick, stronge, and the achenes are nearly cylindrical, dark brown or black, about 3-5 mm in length, and named Kalaziran and Kaliziri in Uyghur medicine［1-2］. The mature achenes of V. anthelmintica is an important medicinal material in traditional medicine, which has the functions of expelling parasite, subsiding swelling, and eliminating cold to stop pain, and is mainly used to treat vitiligo, damp-cold stomach pain, liver disease, inflammation and intestinal parasites［3］. The single medicine and compound preparations developed with V. anthelmintica as the main medicine include injections, ointments, liniments, tablets, pills, etc.［4-6］, among which Compound Quchongbanjiuju Pill and Quchongbanjiuju injection are widely used in clinical practice and have good effects on the treatment of vitiligo［7-8］. As a main medicinal material in the commonly used prescriptions for the treatment of vitiligo in Uyghur medicine, the main anti-vitiligo components isolated from V. anthelmintica are caffeoylquinic acids and sesquiterpenes［9-11］. Our research team established the fingerprint of V. anthelmintica based on HPTLC and HPLC-PDA technology, used HPLC-QTOF-MS to identify the main components of the medicinal material combinining with statistical analysis, and found that the main active ingredients of V. anthelmintica seeds are 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid［10］. According to literatures, V. anthelmintica is distributed in India, Pakistan, Sri Lanka, Nepal, Afghanistan, Myanmar, Laos, Malaysia and Chinas western Yunnan, Xinjiang (Hotan, Aksu, etc.), Guangxi Province, Shaanxi Province, etc.［12］. V. anthelmintica seeds are mainly imported, and there is currently no large-scale production and no normal seed sales channels. The seeds are dormant, and the germination rate of the purchased seeds is usually between 5% and 65%, which hinders the planting and seed production of V. anthelmintica. In order to keep the internal quality and clinical efficacy of V. anthelmintica decoction and various preparations from being affected by the quality of its seeds as a medicinal material, on the basis of the previous research, the effects of seeding rate and light on the seed yield and active ingredients of V. anthelmintica were investigated, aiming to improve the seed yield and quality of V. anthelmintica through the regulation of the seeding rate and light conditions and to further lay a foundation for V. anthelmintica planting and production and anti-vitiligo research.
Materials and Methods
Sources of test seeds
V. anthelmintica seeds were purchased from Kashgar Kunlun Pharmaceutical Co., Ltd. of Xinjiang in 2015. The original medicinal material was identified as the mature seeds of V. anthelmintica in Vernonia of Compositae by Associate Researcher Feng Ying of Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences. The specimen (WY02316) is deposited in the Key Laboratory of Plant Resources and Chemistry of Arid Zone, Chinese Academy of Sciences. We have carried out a standardized planting test of V. anthelmintica in Moyu County, Hotan, Xinjiang since 2015. The seeding material for this test was the self-propagated and self-retained seed material purchased above.
Design of field trials
The experimental area is located in Moyu Agricultural Science and Technology Park, Zawa Town, Moyu County, Hotan Prefecture, at E 79°37′, N 37°09′, and 1 386 m above sea level. The planting area was 1.5 hm2. The annual average temperature in this area is 11.4 ℃; and the accumulated temperature on the limit of above 0 ℃ is 4 450 ℃, and that on the limit equal to or greater than 10 ℃ is 4 017 ℃. The frost-free period is about 210 d. The main source of irrigation water was groundwater. The soil is fertile sandy loam with a pH value of 7.3-7.6. The 1 000-seed weight was 4.22 g, and the total annual irrigation amount was 9 000-12 000 m3/hm2. The experiment adopted an orthogonal experiment design, and set with 9 treatments, in 3 replicates, i.e., 27 plots in total. Seeding was carried out in early April with a planting depth of 1.0-2.0 cm and a row spacing of 40 cm. Based on the previous seeding rate control experiment, the seeding rate of V. anthelmintica was set to 12.0, 13.5 and 15.0 kg/hm2, and the light was divided into two layers of shading (Z2), one layer of shading (Z1), and no shading (CK), as shown in Table 1.
The phenology and plant growth changes of V. anthelmintica were observed every 10 d; during the seed maturiation period of V. anthelmintica, harvesting, drying, and sieving were carried out in stages according to the seed maturity conditions in plots.
Testing methods
Seed yield testing method
Seed yield was measured by the actual cutting method, which is to collect V. anthelmintica seeds and harvest from the harvested stems, then weigh them with scales, water meters, etc., and measure impurities.
Method for determination of active ingredient contents
The contents of 3,4-dicaffeoylquinic acid (3,4-CQA), 3,5-dicaffeoylquinic acid (3,5-CQA), and 4,5-dicaffeoylquinic acid (4,5-CQA) and the content of total caffeoylquinic acid (3 total CQA) in V. anthelmintica seeds were determined by HPLC.
Data processing methods
SPSS16.0 and Originpro9.0 were used for data statistics and analysis.
Results and Analysis
Effect of seeding rate+light on seed yield of V. anthelmintica
V. anthelmintica seeds were matured in stages, generally from mid-to-late August to early October. At first, mature V. anthelmintica seeds in each plot were harvested daily, dried, cleaned and stored separately; by the beginning of October, when the seeds on the plants were basically mature, V. anthelmintica stalks were harvested by plot at one time, and the seeds were collected, dried, cleaned and stored separately. Then, the yields of V. anthelmintica seeds under different treatment conditions were measured, and the relationship between each seeding rate+light treatment and the seed yield of V. anthelmintica is shown in Fig. 1 below.
From Fig. 1 above, it can be seen that there were differences in the seed yield of V. anthelmintica under various seeding rate+shading treatment conditions. The differences in the seed yield of V. anthelmintica under different seeding rate+shading treatment conditions were rather large. Treatment 8 (seeding rate 15.0 kg/hm2+one shading layer) had the highest yield of V. anthelmintica, followed by treatment 7 (seeding rate 15.0 kg/hm2+two shading layers), while treatment 9 (seeding rate 15.0 kg/hm2+no shading) showed the lowest seed yield. Effects of seeding rate+light on the active ingredients of V. anthelmintica
The contents of 3,4-CQA, 3,5-CQA, 4,5-CQA and total CQA of V. anthelmintica seeds under different seeding rates+light conditions were tested and analyzed. The results are shown in Fig. 2 below.
It can be seen from Fig. 2 that under different seeding rate+light treatment conditions, the contents of 3,4-CQA, 3,5-CQA, 4,5-CQA, and total CQA in V. anthelmintica seeds were different, and the contents of 3,5-CQA and total CQA content varied greatly, while the contents of 3,4-CQA and 4,5-CQA varied very little. Specifically, under the condition of treatment 8 (the seeding rate of 15 kg/hm2 and one layer of shading), the 4,5-CQA and total CQA contents of V. anthelmintica seeds were significantly higher than other treatments; treatment 6 (the seeding rate of 13.5 kg/hm2, no shading) showed a higher total CQA content, while the 3,5-CQA content was higher than other treatments; treatment 9 (the seeding rate of 15.0 kg/hm2+no shading) showed a 3,4-CQA content higher than other treatments; under the condition of treatment 2 (the seeding rate of 12.0 kg/hm2+one layer of shading), the contents of 3,4-CQA, 3,5-CQA and total CQA were lower than other treatments; and under the condition of treatment 1 (the seeding rate of 12.0 kg/hm2+two layers of shading), the content of 4,5-CQA was lower than other treatments. It can be seen that the seeding rate and light had important effects on the quality of V. anthelmintica.
Conclusions
After 3 years of seeding density control gradient test in the early stage, we concluded that the suitable seeding rate for V. anthelmintica in Moyu County, Hotan and similar areas was 12.0-15.0 kg/hm2. Based on this, the effects of different seeding rate+light conditions on the seed yield and active ingredients of V. anthelmintica were investigated in this study, and the results showed that the suitable seeding rate of V. anthelmintica was 13.5-15.0 kg/hm2.
① If the purpose is to harvest the seeds of V. anthelmintica, the suitable sowing rate in Moyu County, Hotan and areas with similar climatic conditions can be 15.0 kg/hm2, and proper shading during the vegetative growth period can increase its yield.
② If the purpose is to extract 4,5-CQA and total CQA in V. anthelmintica seeds, the optimal seeding rate and light for planting V. anthelmintica can be 15 kg/hm2 and one layer of shading. For the purpose of 3,5-CQA in V. anthelmintica seeds, the optimal seeding rate and light for planting V. anthelmintica recommended are 13.5 kg/hm2 and no shading. If 3,4-CQA is to be extracted from V. anthelmintica seeds, the optimal seeding rate and light can be 15.0 kg/hm2 and no shading. ③ The seed yield of V. anthelmintica and the contents of 3,4-CQA, 3,5-CQA and total CQA under the condition of 12.0 kg/hm2+one layer of shading were lower than other treatments, while under the condition of the seeding rate 12 kg/hm2+two layers of shading, the content of 4,5-CQA in seeds was lower than other treatments.
Agricultural Biotechnology2021
References
［1］ Chinese Pharmacopoeia Commission. Drug Standards of the Ministry of Health, the Peoples Republic of China-Uyghur Medicines: Uyghur Medicine Section［S］. Urumchi: Xinjiang Science and Technology Health Press, 1998. (in Chinese)
［2］ The state Administration of Traditional Chinese Medicine of Chinese Materia Medica editorial board. Chinese materia medica, Uygur part［M］. Shanghai: Shanghai science and Technology Press, 2005: 206-207. (in Chinese)
［3］ SHI DR, TIAN HY, LIU JL, et al. Preliminary study on the treatment of vitiligo with Uyghur medicine and western medicine［J］. Xinjiang Zhongyiyao, 2005,23(1): 31-32. (in Chinese)
［4］ National Institutes for Food and Drug Control. Records of Chinese ethnodrugs［M］. Beijing: Peoples Medical Publishing House. (in Chinese)
［5］ The Editorial Board of Flora of Xinjiang. Flora of Xinjiang［M］. Urumchi: Xinjiang Science and Technology Health Press, 1999. (in Chinese)
［6］ LIU YM, SAWET YIKIM. Records of Uyghur medicine［M］. Urumchi: Xinjiang Science and Technology Health Press,1993. (in Chinese)
［7］ HURXIGUL MAMAT. Clinical observation on 90 cases of vitiligo treated by Uygur medicine［J］. Jornal of Xlnnjiang Ulghur medical College, 1997(3): 26-31. (in Chinese)
［8］ TURSUN UPUR, ANIWAR JAMAL. Observation on curative effect of uigurian medicine in treating 610 cases suffered from Pityriasis versicolor alba［J］ . Journal of Medicine & Pharmacy of Chinese Minoritie, 2003(1): 12. (in Chinese)
［9］ LIU ZY, ZAN K, WU X, et al. Research progress on chemical constituents, quality control and pharmacological activity of Vernonia anthelmintica［J］. Chinese Journal of Pharmaceutical Analysis, 2020, 40(2): 199-208. (in Chinese)
［10］ MAIMAITI ZULIPIYA. Study on the optimization of the preparation process and quality standards of the active ingredients from Vernonia anthelmintica［D］. Beijing: University of Chinese Academy of Sciences, 2019. (in Chinese)
［11］ ABDUWAR ABDULHAK, SALAMET ELY, WANG YL. Studies on quality control of uyghur crude drug coriander seeds of Vernonia anthelmintica Willd［J］. Journal of Medicine & Pharmacy of Chinese Minoritie, 2015, 21(4): 44-46. (in Chinese)
［12］ L U TH. Research and application of uighur medicine Vernonia anthelmintica in the treatment of vitiligo［J］. Foreign Medical Sciences, 2004, 26(2): 103-104. (in Chinese)