蛹虫草培养残基中虫草素的水热回流提取及含量测定

doi:10.16431/j.cnki.1671-7236.2022.09.007

Abstract

Abstract: 【Objective】 The purpose of this study was to explore the optimal extraction conditions of cordycepin from Cordyceps militaris residual medium by the hydrothermal reflux method.【Method】 The single factor design and the orthogonal experiment design were conducted according to four factors including the liquid to material ratio, extraction repetitions, extraction temperature and extraction time, and the cordycepin content was measured by high-performance liquid chromatography (HPLC) method, which were used to decide the optimal extraction parameters of cordycepin from Cordyceps militaris residual medium by the hydrothermal reflux method.【Result】 The results of methodology showed that the relative standard deviation (RSD) of precision, repeatability, and stability for HPLC detection of cordycepin content in the extract of Cordyceps militaris residual medium were less than 2%, the RSD of accuracy was 2.64%, and the standard curve had high linear correlation.The results of the orthogonal experiment showed that the order of factors affecting the extraction efficacy of the hydrothermal reflux method were extraction time, extraction repetitions, liquid to material ratio and extraction temperature, and extraction time and extraction repetitions had significant effects on cordycepin content in extract (P<0.05), while extraction temperature and liquid to material ratio had not obvious effects (P>0.05).The results of the single factors and the orthogonal experiment design showed that the optimal extraction process factors of cordycepin from Cordyceps militaris residual medium were the ratio of liquid to material 25:1, repetitions 5 times, extraction temperature 70 ℃, and extraction time 60 min, respectively.Under these extraction conditions, the cordycepin content of Cordyceps militaris residual medium was 1.48 mg/g.【Conclusion】 The optimal extraction parameters of the hydrothermal reflux method in this study could effectively extract the active components from Cordyceps militaris residual medium, the operation process was easy and practical, and the yield of the active components of the extract was stable.The results provided reference for the development of product from Cordyceps militaris residual medium.

Key words: Cordyceps militaris residual medium; cordycepin; hydrothermal reflux method; extraction process

CLC Number:

R284.2

PENG Zhiping, ZHANG Jingyan, GUO Zhiting, WANG Lei, ZHANG Kai, ZHANG Kang, WANG Guibo, QIU Zhengying, WANG Xuezhi, LI Jianxi. Extraction and Content Detection of Cordycepin from Cordyceps militaris Residual Medium by Hydrothermal Reflux Method[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3333-3342.

References

[1] 郑壮丽, 黄春花, 梅彩英, 等.蛹虫草国内外研究的新进展[J].环境昆虫学报, 2011, 33(2):225-233. ZHENG Z L, HUANG C H, MEI C Y, et al.Current advance on the research of Cordyceps militaris[J].Journal of Environmental Entomology, 2011, 33(2):225-233.(in Chinese)
[2] 吴征镒, 周太炎, 肖培根, 等.新华本草纲要:第三册[M].上海:上海科学技术出版社, 1990. WU Z Y, ZHOU T Y, XIAO P G, et al.Compendium of Materia Medica of Xinhua:Volume Ⅲ[M].Shanghai:Shanghai Scientific & Technical Publishers, 1990.(in Chinese)
[3] LEE C T, HUANG K S, SHAW J F, et al.Trends in the immunomodulatory effects of Cordyceps militaris:Total extracts, polysaccharides and cordycepin[J].Frontiers in Pharmacology, 2020, 11:575704.
[4] OLATUNJI O J, TANG J, TOLA A, et al.The genus Cordyceps:An extensive review of its traditional uses, phytochemistry and pharmacology[J].Fitoterapia, 2018, 129:293-316.
[5] DAS S K, MASUDA M, SAKURAI A, et al.Medicinal uses of the mushroom Cordyceps militaris:Current state and prospects[J].Fitoterapia, 2010, 81(8):961-968.
[6] 文庭池, 查岭生, 康冀川, 等.蛹虫草研究和开发过程中的一些问题和展望[J].菌物学报, 2017, 36(1):14-27. WEN T C, ZHA L S, KANG J C, et al.Problems and prospects of research and development of Cordyceps militaris[J].Mycosystema, 2017, 36(1):14-27.(in Chinese)
[7] 陈丽冰.北虫草固体培养残基中多糖和虫草素高效提取与纯化研究[D].荆州:长江大学, 2015. CHEN L B.High efficiency extraction and purification of polysaccharide and cordycepin from Cordyceps militaris reside medium[D].Jingzhou:Yangtze University, 2015.(in Chinese)
[8] VUONG H T, NGUYEN C P, PHAN L T M, et al.Ultrasound-assisted enzymatic extraction of adenosine from Vietnamese Cordyceps militaris and bioactivity analysis of the extract[J].Journal of Chemistry, 2020, 2020:1-10.
[9] 赵星月, 李倩, 刘文静, 等.蛹虫草菌生物合成虫草素的研究进展[J].生物工程学报, 2020, 36(7):1293-1304. ZHAO X Y, LI Q, LIU W J, et al.Advances in biosynthesis of cordycepin from Cordyceps militaris[J].Chinese Journal of Biotechnology, 2020, 36(7):1293-1304.(in Chinese)
[10] 王英娟, 李多伟, 王义潮, 等.蛹虫草中虫草素、虫草多糖综合提取工艺研究[J].西北植物学报, 2005, 25(9):1863-1867. WANG Y J, LI D W, WANG Y C, et al.Integrated extracting technology of cordycepin and poly-saccharides in Cordyceps militaris[J].Acta Botanica Boreali-Occidentalia Sinica, 2005, 25(9):1863-1867.(in Chinese)
[11] 雷燕妮, 张小斌, 陈书存, 等.蛹虫草菌渣中虫草素、多糖、多肽综合提取分离的工艺研究[J].陕西农业科学, 2021, 67(9):32-37. LEI Y N, ZHANG X B, CHEN S C, et al.Study on comprehensive extraction and separation of cordycepin, polysaccharide and peptide from Cordyceps militaris residue[J].Shaanxi Journal of Agricultural Sciences, 2021, 67(9):32-37.(in Chinese)
[12] AZWANIDA N N.A review on the extraction methods use in medicinal plants, principle, strength and limitation[J].Medicinal and Aromatic Plants, 2015, 4(3):1000196.
[13] 邢增涛, 饶钦雄, 黄志英, 等.虫草制品中虫草素和腺苷的测定高效液相色谱法:NY/T 2116-2012[S].北京:中国人民共和国农业部, 2012. XING Z T, RAO Q X, HUANG Z Y, et al.Determination of cordycepin and adenosine in Cordyceps products by high performance liquid chromatography method:NY/T 2116-2012[S].Beijing:Ministry of Agriculture of the People's Republic of China, 2012.(in Chinese)
[14] 国家药典委员会.中华人民共和国药典:四部[M].北京:中国医药科技出版社, 2020. CHINESE PHARMACOPOEIA COMMISSION.Pharmacopoeia of the People's Republic of China:Volume Ⅳ[M].Beijing:China Medical Science Press, 2020.(in Chinese)
[15] PINTATHONG P, CHOMNUNTI P, SANGTHONG S, et al.The feasibility of utilizing cultured Cordyceps militaris residues in cosmetics:Biological activity assessment of their crude extracts[J].Journal of Fungi, 2021, 7(11):973.
[16] NI H, ZHOU X, LI H, et al.Column chromatographic extraction and preparation of cordycepin from Cordyceps militaris waster medium[J].Journal of Chromatography B, 2009, 877(22):2135-2141.
[17] 余俊杰.蛹虫草菌株鉴定及虫草素提取工艺优化和产品研发[D].合肥:安徽农业大学, 2021. YU J J.Identification of Cordyceps militaris strains and optimization of cordycepin extraction process and product development[D].Hefei:Anhui Agricultural University, 2021.(in Chinese)
[18] 吕丹, 郑雪, 许登泉, 等.UV法和HPLC法测定北虫草中虫草素含量的方法比较[J].广东化工, 2013, 40(16):201-202. LYU D, ZHENG X, XU D Q, et al.Comparison of UV and HPLC methods for the determination of the content of cordycepin in Cordyceps militaris[J].Guangdong Chemical Industry, 2013, 40(16):201-202.(in Chinese)
[19] 李剑梅, 郝捷, 池景良, 等.响应面法优化蛹虫草子实体虫草素提取工艺[J].中国食用菌, 2021, 40(5):62-68. LI J M, HAO J, CHI J L, et al.Optimization of extraction of cordycepin from fruiting body of Cordyceps militaris by response surface methodology[J].Edible Fungi of China, 2021, 40(5):62-68.(in Chinese)
[20] TULI H S, SHARMA A K, SANDHU S S, et al.Cordycepin:A bioactive metabolite with therapeutic potential[J].Life Sciences, 2013, 93(23):863-869.
[21] ZHANG J X, WEN C T, DUAN Y Q, et al.Advance in Cordyceps militaris (Linn.) Link poly-saccharides:Isolation, structure and bioactivities:A review[J].International Journal of Biological Macromolecules, 2019, 132:906-914.
[22] LUO Q, CAO H, LIU S, et al.Novel liquid fermentation medium of Cordyceps militaris and optimization of hydrothermal reflux extraction of cordycepin[J].Journal of Asian Natural Products Research, 2020, 22(2):167-178.
[23] 张德斌, 翟文姣, 刘国香, 等.不同粒径对蛹虫草活性成分溶出量的影响[J].食品科技, 2021, 46(12):204-209. ZHANG D B, ZHAI W J, LIU G X, et al.Effects of different particle sizes on dissolution of the active ingredients from Cordyceps militaris[J].Food Science and Technology, 2021, 46(12):204-209.(in Chinese)
[24] 常正姣.杂粮蛹虫草菌丝共生体的培养工艺参数优化[D].郑州:郑州大学, 2016. CHANG Z J.Parameter optimization of culture processing of multigrain Cordyceps militaris hyphae symbionts[D].Zhengzhou:Zhengzhou University, 2016.(in Chinese)
[25] 孟胜楠.蛹虫草子座培养剩余基质中虫草素的提取及分离[D].杨凌:西北农林科技大学, 2013. MENG S N.Extraction and separation of cordycepin from Cordyceps militaris medium[D].Yangling:Northwest A&F University, 2013.(in Chinese)
[26] 张丽艳.从蛹虫草栽培残渣中提取分离虫草素及其抑制肿瘤活性的研究[D].福州:福建师范大学, 2010. ZHANG L Y.Extraction and isolation of cordycepin from the deserted solid medium of Cordyceps militaris and antitumor effects[D].Fuzhou:Fujian Normal University, 2010.(in Chinese)
[27] 石浩.蛹虫草下脚料主要活性成分提取及其对西葫芦保鲜贮藏的研究[D].无锡:江南大学, 2019. SHI H.Extraction of main active components from the spent substrate of Cordyceps militaris and its application in fresh storage of Cucurbita pepo L.[D].Wuxi:Jiangnan University, 2019.(in Chinese)

Extraction and Content Detection of Cordycepin from Cordyceps militaris Residual Medium by Hydrothermal Reflux Method

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 12

Recommended Articles

Metrics

Comments

[1]	LIU Peng, LI Jingjing, QIAO Yanliang, ZHU Lianqin, ZHU Fenghua. Extraction and Structure Characterization of Scutellaria baicalensis Polysaccharide [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2518-2530.
[2]	LI Yahui, LUO Shengjun, TANG Xinggang, YUAN Minggui, WEI Qilin, LOU Hua, XIANG Rong. Study on Optimization of Extraction Process and Antibacterial Activity to Clostridium Perfringens of Total Flavonoids from Vitex negundo L.var. cannabifolia (Sieb.et Zucc.) Hand.-Mazz. [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2406-2413.
[3]	CHEN Yuxiao, LI Huanjuan, XI Xiaohua, HAO Jie, ZHOU Degang, SI Hongbin. Study on Extraction Process of Jiawei Pingwei Oral Liquid [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2414-2422.
[4]	YAN Lijun, LYU Xinyu, ZHANG Duo, CHEN Changbin, ZHANG Shuang, GAO Yuan, LUO Jianghan, WANG Fuling, CUI Wenyu, ZHANG Chengxian. Extraction of Effective Fraction of Huanglian Jiedu Decoction and Its Scavenging Effect on Candida albicans in vivo [J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(11): 3414-3421.
[5]	LI Yan, XU Hongyue, LI Shulin, WANG Xueyan, TANG Xudong, YU Lu. Study on Cordycepin Prevents Obesity and Mediates Prolactin Secretion Through ADORA₁ in GH3 Cells [J]. , 2018, 45(5): 1382-1388.
[6]	XIE Kan, XIONG Chun-mei, FANG Li-yuan, JIANG Chan, LEI Fu-hong, OUYANG Wu-le, GUO Ai-wei. Optimization of Oil Extraction Process from Jatropha curcas Seed and the Analysis of Crude Protein Content in Meal [J]. , 2017, 44(10): 2951-2956.
[7]	LU Guang-fu, CHEN Xiao-lan, HUANG Ya-qi, JIANG Chun-mao, YU Jie, QU Jing-wen, GENG Qian-li. Study on the Optimum Extraction Process of Mulberry Leaf Polysaccharide [J]. , 2016, 43(7): 1922-1927.
[8]	WANG Rui, CHU Xiu-ling, SU Jian-qing, ZHANG Min, DING Chan. Optimization of Stachydrine Hydrochloride in Leonurus Extraction Method by HPLC-ELSD [J]. , 2016, 43(3): 707-713.
[9]	SUN Huan-lin，ZHANG Wen-ju，LIU Yan-feng，TANG Shu-zhen. Biological Function and Extraction Process of Glycyrrhizic Acid and its Application in Monogastric Animal [J]. , 2014, 41(7): 126-130.
[10]	GU Yu-feng, HUANG Ya-jun, WANG Qin, WU Yong, LIU Zhao-ying, ZHANG Ming-jun, SUN Zhi-liang, JIANG Zheng-yun. Study on the Extraction Process of Compound Epimedium by Orthogonal Experimental Design [J]. , 2014, 41(3): 255-258.
[11]	LI Xiao-zhen, JIA Shu-hong, ZHANG Dong-sheng, ZHU Xiao-qing, LUO Yan, SUN Yue-bo, CHEN Jie, GU Xin-li. Study on Extraction Process of Alkaloids in Traditional Chinese Medicine Compound Gongyanjing [J]. , 2014, 41(10): 113-117.
[12]	XIONG Chun-mei;GUO Ai-wei;CHEN Fen-fen;WANG San-mei. Experimental Study on Parameter Optimization of Extraction Process of Leaf Protein from Clover [J]. , 2011, 38(12): 61-63.