องค์ประกอบทางเคมี และฤทธิ์ต้านอนุมูลอิสระของสารสกัดถั่งเช่าสีทอง (Cordyceps militaris) ที่เพาะเลี้ยงในอาหารเพาะเหลว และอาหารเพาะแข็งธัญพืชลูกเดือย
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ม.ค. 21, 2021
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ถั่งเช่าสีทอง อาหารเพาะ องค์ประกอบทางเคมี ฤทธิ์ต้านอนุมูลอิสระ
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บทคัดย่อ
งานวิจัยนี้ มีวัตถุประสงค์เพื่อศึกษาองค์ประกอบทางเคมี และฤทธิ์ต้านอนุมูลอิสระของสารสกัดถั่งเช่าสีทองที่เพาะเลี้ยงในอาหารเพาะเหลว และที่เพาะเลี้ยงในอาหารเพาะแข็งธัญพืชลูกเดือย วิเคราะห์องค์ประกอบทางเคมีด้วยวิธี GC-MS และศึกษาฤทธิ์ต้านอนุมูลอิสระโดยวิเคราะห์ฟีนอลิกและฟลาโวนอยด์รวม และวิเคราะห์ความสามารถในการต้านอนุมูลอิสระโดยวิธี DPPH และ ABTS assay ผลการทดลอง พบสารในสารสกัดถั่งเช่าสีทองที่เพาะเลี้ยงในอาหารเพาะเหลว 23 ชนิด ที่เพาะเลี้ยงในอาหารเพาะแข็งธัญพืชลูกเดือย 26 ชนิด มีสารเหมือนกันในสารสกัดทั้งสอง 16 ชนิด สารหลักที่พบในสารสกัดทั้งสองชนิด คือ คอร์ไดซีปิน ปริมาณฟีนอลิกและฟลาโวนอยด์รวมของสารสกัดถั่งเช่าสีทองที่เพาะเลี้ยงในอาหารเพาะเหลว คือ 178.009±0.712 mgGE/gExt และ 91.000±0.953 mgQE/gExt ซึ่งสูงกว่าที่เพาะเลี้ยงในอาหารเพาะแข็งธัญพืชลูกเดือย (135.48+0.602 mgGE/gExt และ 6.817±0.195 mgQE/gExt) ความสามารถในการกำจัดอนุมูลอิสระ DPPH และ ABTS ของสารสกัดถั่งเช่าสีทองที่เพาะเลี้ยงในอาหารเพาะเหลว มีค่า IC50 เป็น 0.499±0.0421 และ 0.068±0.004 mg/mL สูงกว่าที่เพาะเลี้ยงในอาหารเพาะแข็งธัญพืชลูกเดือย (0.995±0.0689 และ 0.223±0.013 mg/mL)
ผลจากการวิจัยครั้งนี้ แสดงให้เห็นว่า สารสกัดถั่งเช่าสีทองที่เพาะเลี้ยงในอาหารเพาะเหลว มีฤทธิ์ต้านอนุมูลอิสระสูงกว่าสารสกัดถั่งเช่าสีทองที่เพาะเลี้ยงในอาหารเพาะแข็งธัญพืชลูกเดือย ฟีนอลิกและฟลาโวนอยด์รวมในสารสกัดทำหน้าที่เป็นสารออกฤทธ์ต้านอนุมูลอิสระ
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References
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2. Hur H. Chemical ingredients of Cordyceps militaris. Microbiol 2008;36(4):233-235.
3. Sung GH, Hywel-Jones NL, Sung JM, Luangsa-ard JJ, Shrestha B, Spatafora JW. Phylogenetic classification of Cordyceps and the Clavicipitaceous fungi. Stud Mycol 2007;57:5-59.
4. Park JG, Son YJ, Lee TH, Baek NJ, Yoon DH, Kim TW, Aravinthan A, Hong S, Kim JH, Sung GH, Cho JY. Anticancer efficacy of Cordyceps militaris ethanol extract in a xenografted leukemia model, Hindawi Evidence-Based Complem Alt Med 2017. https://DOI.org/10.1155/2017/8474703.
5. Gui ZZ, Zhu YH. Advance on cultivation, bioactive compound and pharmacological mechanism of Cordyceps militaris. Sci Seri 2008;34:178-184.
6. Zheng P, Xia Y, Xiao G, Xiong C, Hu X, Zhang S, Zheng H, Huang Y, Zhou Y, Wang S et al. Genome sequence of the insect pathogenic fungus Cordyceps militaris, a valued traditional Chinese medicine. Genome Biol 2011;(12):116.
7. Dong CH, Yang T, Lian T. A Comparative study of the antimicrobial, antioxidant, and cytotoxic activities of methanol extracts from fruit bodies and fermented mycelia of caterpillar medicinal mushroom Cordyceps militaris (Ascomycetes). Int J Med Mushrooms 2014;16(5):485-495 DOI: 10.1615/Int J Med Mushrooms.v16.i5.70
8. Dai JJ, Fan T, Wu CH, Xiao LZ, Tian SF. Summarization of the study on the artificial cultivation of Cordyceps militaris Link. J Anhui Agric Sci 2007;35:5469–5471.
9. Paterson R. Cordyceps: a traditional Chinese medicine and another fungal therapeutic biofactory. Phytochem 2008; 69:1469-1495.
10. Cui J.D. Biotechnological production and applications of Cordyceps militaris, a valued traditional Chinese medicine. Crit Rev Biotechnol 2014;35:475-484.
11. Mizuno T. Medicinal effects and utilization of Cordyceps (Fr.) Link (Ascomycetes) and Isaria Fr. (Mitosporic Fungi) Chineese caterpiller fungi, “Tochukaso” (Review). Int J Med Mushroom 1999;1:251-261.
12. Ng TB, Wang HX. Pharmacological actions of Cordyceps, a prized folk medicine. J Pharm Pharmacol 2005;57:1509- 1519.
13. Rukachaisirikul V, Pramjit S, Pakawatchai C, Isaka M, Supothina S. 10-Membered macrolides from the insect pathogenic fungus Cordyceps militaris BCC 2816. J Nat Prod 2004;67:1953-1955.
14. Hur H. Chemical ingredients of Cordyceps militaris Microbiol 2008;36(4):233-235.
15. Reis FS, Barros L, Calhelha RC, Cirić A, van Griensven LJ, Soković M, Ferreira IC. The methanolic extract of Cordyceps militaris (L.) Link fruiting body shows antioxidant, antibacterial, antifungal and antihuman tumor cell lines properties. Food Chem Toxicol 2013;62:91-8. DOI:10.1016/j.fct.2013.08.033.
16. Cunningham KG, Hutchinson SA, Manson W, Spring FS. Cordycepin, a metabolic product from cultures of Cordyceps militaris (Linn.) Link. Part I. Isolation and characterization. J Chem Soc 1951;2299–3200.
17. Yu HM, Wan BS, Huang SC, Duh PD. Comparison of protective effects between cultured Cordyceps militaris and natural Cordyceps sinensis against oxidative damage. J Agric Food Chem 2006;54: 3132-3138.
18. Huang L, Li Q, Chen Y, Wang X, Zhou X. Determination and analysis of cordycepin and adenosine in the products of Cordyceps spp. Afri J Microbiol Res 2009; 3 (12):957-961. http://www.academic journals.org/ajmr
19. Chou SM, Lai WJ, Hong TW, Lai JY, Tsai SH, Chen YH, Yu SH, Kao CH, Chu R, Ding ST, Li TK, Shen TL. Synergistic property of cordycepin in cultivated Cordyceps militaris-mediated apoptosis in human leukemia cells. Phytomed 2014;21: 1516-1524.
20. Ahn YJ, Park SJ, Lee SG, Shin SC, Choi DH. Cordycepin: selective growth inhibitor derived from liquid culture of Cordyceps militaris against Clostridium spp. J Agric Food Chem 2002;48:2744-2748.
21. Nakamura K, Shinozuka K, Yoshikawa N. Anticancer and antimetastatic effects of cordycepin, an active component of Cordyceps sinensis. J Pharmacol Sci 2015; 127:53-56.
22. Jo WS, Choi YJ, Kim HJ, Lee JY, Nam BH, Lee JD, Lee SW, Seo SY, Jeong MH. The anti-inflammatory effects of water extract from Cordyceps militaris in murine macrophage. Mycobiol 2010;38(1):46-51. DOI:10.4489/MYCO.2010.38.1.046.
23. Jeong MH, Park YS, Jeong DH, Lee CG, Kim JS, Oh SJ, JeongSK, Yang K, Jo WS. In vitro evaluation of Cordyceps militaris as a potential radioprotective agent. Int J Molec Med 2014;34:1349-1357. DOI: 10.3892/ijmm.2014.1901
24. Tuli HS, Sharma AK, Sandhu SS, Kashyap D. Cordycepin: a bioactive metabolite with therapeutic potential. Life Sci 2013;93:863-869.
25. Shrestha B, Zhang W, Zhang Y, Liu X. The medicinal fungus Cordyceps militaris: research and development. Mycol Prog 2012;11:599-614.
26. Quy TN, Xuan TD, Andriana Y, Tran HD, Khanh TD, Teschke R. Cordycepin Isolated from Cordyceps militaris: Its newly discovered herbicidal property and potential plant-based novel alternative to glyphosate. Molec 2019;24:2901.DOI:10.3390/molecules24162901 www.mdpi.com/journal/molecules
27. Wasser SP. Medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides. Appl Microbiol Biotechnol 2002;60:258-274.
28. Liu F, Ooi VEC, Chang ST. Free radical scavenging activities of mushroom polysaccharide extracts. Life Sci 1997;60: 763-771.
29. Wu F, Yan H, Ma X, Jia J, Zhang G, Guo X, Gui Z. Structural characterization and antioxidant activity of purified polysaccharide from cultured Cordyceps militaris. Afr J Microbiol Res 2011;5(18): 2743-2751. http://www.academicjournals.org/ajmr DOI:10.5897/AJMR11.548
30. Yoo HS, Shin JW, Cho JH, et al. Effects of Cordyceps militaris extract on angiogenesis and tumor growth. Acta Pharmacol Sin 2004;25:657-665.
31. Won SY, Park EH. Anti-inflammatory and related pharmacological activities of cultured mycelia and fruiting bodies of Cordyceps militaris. J Ethnopharmacol 2005;96:555-561.
32. Yu HM, Wang BS, Huang SC, Duh PD. Comparison of protective effects between cultured Cordyceps militaris and natural Cordyceps sinesis against oxidative damage. J Agric Food Chem 2006;54: 3132-3138.
33. Chen C, Luo SS, Li Y, Sun YJ, Zhang CK. Study on antioxidant activity of three Cordyceps sp. by chemiluminescence. Shanghai J Trad Chinese Med 2004;38: 53-55.
34. Wu WC, Hsiao JR, Lian YY, Lin CY, Huang BM. The apoptotic effect of cordycepin on human OEC-M1 oral cancer cell line. Cancer Chemother Pharmacol 2007;60: 103-111.
35. Lin YW, Chiang BH. Anti-tumor activity of the fermentation broth of Cordyceps militaris cultured in the medium of Radix astragali. Proc Biochim 2008;43:244-250.
36. Wei HP, Xiao B, Hu KZ. Pharmaceutical values of Cordyceps militaris. J Chin Med Mater 2004;27:215-217.
37. Patel KJ, Ingalhalli RS. Cordyceps militaris (L.: Fr.) Link – An important medicinal mushroom. J Pharmacog Phytochem 2013; 2:315. www.phytojournal.com
38. Oh J, Yoon DH, Shrestha B, Choi HK, Sung GH. Metabolomic profiling reveals enrichment of cordycepin in senescence- process of Cordyceps militaris fruit bodies. J Microbiol 2019;57(1):54-63.DOI: 10.1007/s12275-019-8486-z
39. Singleton VL, Orthofer RLR. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods Enzym 1999; 299:152-178.
40. Chang C, Yang M, Wen HCJ. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 2002;10:178-182.
41. Ursini F, Maiorino M, Morazzoni P, Roveri A PG. A novel antioxidant flavonoid (IdB 1031) affecting molecular mechanisms of cellular activation. Free Radic Biol Med 1994;16:547-553.
42. Lee HL, Halliwell B. Oxidation and generation of hydrogen peroxide by thiol compounds in commonly used cell culture media. Biochem Biophys Res Commun 2001;286:991-994.
43. Guo C, Zhu J, Zhang C, Zhang LJ. Determination of adenosine and 3'-deoxyadenosine in Cordyceps militaris (L.) Link. by HPLC. J Chinese Med 1998;23: 236-237.
44. Jung EC, Kim KD, Bai CH, Kim JC, Kim DK, Kim HH. A mushroom lectin from ascomycete Cordyceps militaris. Biochem Biophys Acta Gen Subj 2007;1770:833-838.
45. Sun YJ, Lu P, Ling JY, Zhang HX, Chen C, Zhang CK. Nucleoside from Cordyceps kyushuensis and the distribution of two active components in its different parts. Yao Xue Xue Bao 2003;38:690–694.
46. Fan Z. Coix: Chemical composition and health effects. Trends Food Sci & Technol 2017;61:160-175.
47. Kondo Y, Nakajima K, Nozoe S, Suzuki S. Isolation of ovulatory-active substances from crops of Job's Tears (Coix lacryma-jobi L. var. ma-yuen Stapf.). Chem Pharm Bull 1988;36:3147-152.
48. Nagao T, Otsuka H, Kohda H, Sato T, Yamasaki K. Benzoxazinones from Coix lachryma-jobi var. ma-yuen. Phytochem 1985;24:2959-2962.
49. Gu YX, Wang ZS, Li SX, Yuan QS. Effect of multiple factors on accumulation of nucleosides and bases in Cordyceps militaris. Food Chem 2007;102:1304-1309.
50. Shih IL, Tsai KL, Hsieh C. Effects of culture conditions on the mycelial growth and bioactive metabolite production in submerged culture of Cordyceps militaris. Biochem Eng J 2007;33:193-201.
51. Dziezak JD. Antioxidants - the ultimate answer to oxidation. Food Technol 1986; 40:94.
52. Mei H, Wang YS, Huang SC, Duh PD. Comparison of protective effects between cultured Cordyceps militaris and natural Cordyceps sinensis against oxidative damage. J Agric Food Chem 2006;54(8): 3132-3138. https://DOI.org/10.1021/jf053111w
53. Joshi M, Sagar A, Singh zkanwar S and Singh S. Anticancer, antibacterial and antioxidant activities of Cordyceps militaris, Ind J Experimental Biol 2019;57:15-20.http://nopr.
niscair.res.in/handle/123456789/ 45567
54. Li SP, Li P, Dong TT, Tsim KW. Anti-oxidation activity of different types of natural Cordyceps sinensis and cultured Cordyceps mycelia. Phytomed 2001;8:207-212.
2. Hur H. Chemical ingredients of Cordyceps militaris. Microbiol 2008;36(4):233-235.
3. Sung GH, Hywel-Jones NL, Sung JM, Luangsa-ard JJ, Shrestha B, Spatafora JW. Phylogenetic classification of Cordyceps and the Clavicipitaceous fungi. Stud Mycol 2007;57:5-59.
4. Park JG, Son YJ, Lee TH, Baek NJ, Yoon DH, Kim TW, Aravinthan A, Hong S, Kim JH, Sung GH, Cho JY. Anticancer efficacy of Cordyceps militaris ethanol extract in a xenografted leukemia model, Hindawi Evidence-Based Complem Alt Med 2017. https://DOI.org/10.1155/2017/8474703.
5. Gui ZZ, Zhu YH. Advance on cultivation, bioactive compound and pharmacological mechanism of Cordyceps militaris. Sci Seri 2008;34:178-184.
6. Zheng P, Xia Y, Xiao G, Xiong C, Hu X, Zhang S, Zheng H, Huang Y, Zhou Y, Wang S et al. Genome sequence of the insect pathogenic fungus Cordyceps militaris, a valued traditional Chinese medicine. Genome Biol 2011;(12):116.
7. Dong CH, Yang T, Lian T. A Comparative study of the antimicrobial, antioxidant, and cytotoxic activities of methanol extracts from fruit bodies and fermented mycelia of caterpillar medicinal mushroom Cordyceps militaris (Ascomycetes). Int J Med Mushrooms 2014;16(5):485-495 DOI: 10.1615/Int J Med Mushrooms.v16.i5.70
8. Dai JJ, Fan T, Wu CH, Xiao LZ, Tian SF. Summarization of the study on the artificial cultivation of Cordyceps militaris Link. J Anhui Agric Sci 2007;35:5469–5471.
9. Paterson R. Cordyceps: a traditional Chinese medicine and another fungal therapeutic biofactory. Phytochem 2008; 69:1469-1495.
10. Cui J.D. Biotechnological production and applications of Cordyceps militaris, a valued traditional Chinese medicine. Crit Rev Biotechnol 2014;35:475-484.
11. Mizuno T. Medicinal effects and utilization of Cordyceps (Fr.) Link (Ascomycetes) and Isaria Fr. (Mitosporic Fungi) Chineese caterpiller fungi, “Tochukaso” (Review). Int J Med Mushroom 1999;1:251-261.
12. Ng TB, Wang HX. Pharmacological actions of Cordyceps, a prized folk medicine. J Pharm Pharmacol 2005;57:1509- 1519.
13. Rukachaisirikul V, Pramjit S, Pakawatchai C, Isaka M, Supothina S. 10-Membered macrolides from the insect pathogenic fungus Cordyceps militaris BCC 2816. J Nat Prod 2004;67:1953-1955.
14. Hur H. Chemical ingredients of Cordyceps militaris Microbiol 2008;36(4):233-235.
15. Reis FS, Barros L, Calhelha RC, Cirić A, van Griensven LJ, Soković M, Ferreira IC. The methanolic extract of Cordyceps militaris (L.) Link fruiting body shows antioxidant, antibacterial, antifungal and antihuman tumor cell lines properties. Food Chem Toxicol 2013;62:91-8. DOI:10.1016/j.fct.2013.08.033.
16. Cunningham KG, Hutchinson SA, Manson W, Spring FS. Cordycepin, a metabolic product from cultures of Cordyceps militaris (Linn.) Link. Part I. Isolation and characterization. J Chem Soc 1951;2299–3200.
17. Yu HM, Wan BS, Huang SC, Duh PD. Comparison of protective effects between cultured Cordyceps militaris and natural Cordyceps sinensis against oxidative damage. J Agric Food Chem 2006;54: 3132-3138.
18. Huang L, Li Q, Chen Y, Wang X, Zhou X. Determination and analysis of cordycepin and adenosine in the products of Cordyceps spp. Afri J Microbiol Res 2009; 3 (12):957-961. http://www.academic journals.org/ajmr
19. Chou SM, Lai WJ, Hong TW, Lai JY, Tsai SH, Chen YH, Yu SH, Kao CH, Chu R, Ding ST, Li TK, Shen TL. Synergistic property of cordycepin in cultivated Cordyceps militaris-mediated apoptosis in human leukemia cells. Phytomed 2014;21: 1516-1524.
20. Ahn YJ, Park SJ, Lee SG, Shin SC, Choi DH. Cordycepin: selective growth inhibitor derived from liquid culture of Cordyceps militaris against Clostridium spp. J Agric Food Chem 2002;48:2744-2748.
21. Nakamura K, Shinozuka K, Yoshikawa N. Anticancer and antimetastatic effects of cordycepin, an active component of Cordyceps sinensis. J Pharmacol Sci 2015; 127:53-56.
22. Jo WS, Choi YJ, Kim HJ, Lee JY, Nam BH, Lee JD, Lee SW, Seo SY, Jeong MH. The anti-inflammatory effects of water extract from Cordyceps militaris in murine macrophage. Mycobiol 2010;38(1):46-51. DOI:10.4489/MYCO.2010.38.1.046.
23. Jeong MH, Park YS, Jeong DH, Lee CG, Kim JS, Oh SJ, JeongSK, Yang K, Jo WS. In vitro evaluation of Cordyceps militaris as a potential radioprotective agent. Int J Molec Med 2014;34:1349-1357. DOI: 10.3892/ijmm.2014.1901
24. Tuli HS, Sharma AK, Sandhu SS, Kashyap D. Cordycepin: a bioactive metabolite with therapeutic potential. Life Sci 2013;93:863-869.
25. Shrestha B, Zhang W, Zhang Y, Liu X. The medicinal fungus Cordyceps militaris: research and development. Mycol Prog 2012;11:599-614.
26. Quy TN, Xuan TD, Andriana Y, Tran HD, Khanh TD, Teschke R. Cordycepin Isolated from Cordyceps militaris: Its newly discovered herbicidal property and potential plant-based novel alternative to glyphosate. Molec 2019;24:2901.DOI:10.3390/molecules24162901 www.mdpi.com/journal/molecules
27. Wasser SP. Medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides. Appl Microbiol Biotechnol 2002;60:258-274.
28. Liu F, Ooi VEC, Chang ST. Free radical scavenging activities of mushroom polysaccharide extracts. Life Sci 1997;60: 763-771.
29. Wu F, Yan H, Ma X, Jia J, Zhang G, Guo X, Gui Z. Structural characterization and antioxidant activity of purified polysaccharide from cultured Cordyceps militaris. Afr J Microbiol Res 2011;5(18): 2743-2751. http://www.academicjournals.org/ajmr DOI:10.5897/AJMR11.548
30. Yoo HS, Shin JW, Cho JH, et al. Effects of Cordyceps militaris extract on angiogenesis and tumor growth. Acta Pharmacol Sin 2004;25:657-665.
31. Won SY, Park EH. Anti-inflammatory and related pharmacological activities of cultured mycelia and fruiting bodies of Cordyceps militaris. J Ethnopharmacol 2005;96:555-561.
32. Yu HM, Wang BS, Huang SC, Duh PD. Comparison of protective effects between cultured Cordyceps militaris and natural Cordyceps sinesis against oxidative damage. J Agric Food Chem 2006;54: 3132-3138.
33. Chen C, Luo SS, Li Y, Sun YJ, Zhang CK. Study on antioxidant activity of three Cordyceps sp. by chemiluminescence. Shanghai J Trad Chinese Med 2004;38: 53-55.
34. Wu WC, Hsiao JR, Lian YY, Lin CY, Huang BM. The apoptotic effect of cordycepin on human OEC-M1 oral cancer cell line. Cancer Chemother Pharmacol 2007;60: 103-111.
35. Lin YW, Chiang BH. Anti-tumor activity of the fermentation broth of Cordyceps militaris cultured in the medium of Radix astragali. Proc Biochim 2008;43:244-250.
36. Wei HP, Xiao B, Hu KZ. Pharmaceutical values of Cordyceps militaris. J Chin Med Mater 2004;27:215-217.
37. Patel KJ, Ingalhalli RS. Cordyceps militaris (L.: Fr.) Link – An important medicinal mushroom. J Pharmacog Phytochem 2013; 2:315. www.phytojournal.com
38. Oh J, Yoon DH, Shrestha B, Choi HK, Sung GH. Metabolomic profiling reveals enrichment of cordycepin in senescence- process of Cordyceps militaris fruit bodies. J Microbiol 2019;57(1):54-63.DOI: 10.1007/s12275-019-8486-z
39. Singleton VL, Orthofer RLR. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods Enzym 1999; 299:152-178.
40. Chang C, Yang M, Wen HCJ. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 2002;10:178-182.
41. Ursini F, Maiorino M, Morazzoni P, Roveri A PG. A novel antioxidant flavonoid (IdB 1031) affecting molecular mechanisms of cellular activation. Free Radic Biol Med 1994;16:547-553.
42. Lee HL, Halliwell B. Oxidation and generation of hydrogen peroxide by thiol compounds in commonly used cell culture media. Biochem Biophys Res Commun 2001;286:991-994.
43. Guo C, Zhu J, Zhang C, Zhang LJ. Determination of adenosine and 3'-deoxyadenosine in Cordyceps militaris (L.) Link. by HPLC. J Chinese Med 1998;23: 236-237.
44. Jung EC, Kim KD, Bai CH, Kim JC, Kim DK, Kim HH. A mushroom lectin from ascomycete Cordyceps militaris. Biochem Biophys Acta Gen Subj 2007;1770:833-838.
45. Sun YJ, Lu P, Ling JY, Zhang HX, Chen C, Zhang CK. Nucleoside from Cordyceps kyushuensis and the distribution of two active components in its different parts. Yao Xue Xue Bao 2003;38:690–694.
46. Fan Z. Coix: Chemical composition and health effects. Trends Food Sci & Technol 2017;61:160-175.
47. Kondo Y, Nakajima K, Nozoe S, Suzuki S. Isolation of ovulatory-active substances from crops of Job's Tears (Coix lacryma-jobi L. var. ma-yuen Stapf.). Chem Pharm Bull 1988;36:3147-152.
48. Nagao T, Otsuka H, Kohda H, Sato T, Yamasaki K. Benzoxazinones from Coix lachryma-jobi var. ma-yuen. Phytochem 1985;24:2959-2962.
49. Gu YX, Wang ZS, Li SX, Yuan QS. Effect of multiple factors on accumulation of nucleosides and bases in Cordyceps militaris. Food Chem 2007;102:1304-1309.
50. Shih IL, Tsai KL, Hsieh C. Effects of culture conditions on the mycelial growth and bioactive metabolite production in submerged culture of Cordyceps militaris. Biochem Eng J 2007;33:193-201.
51. Dziezak JD. Antioxidants - the ultimate answer to oxidation. Food Technol 1986; 40:94.
52. Mei H, Wang YS, Huang SC, Duh PD. Comparison of protective effects between cultured Cordyceps militaris and natural Cordyceps sinensis against oxidative damage. J Agric Food Chem 2006;54(8): 3132-3138. https://DOI.org/10.1021/jf053111w
53. Joshi M, Sagar A, Singh zkanwar S and Singh S. Anticancer, antibacterial and antioxidant activities of Cordyceps militaris, Ind J Experimental Biol 2019;57:15-20.http://nopr.
niscair.res.in/handle/123456789/ 45567
54. Li SP, Li P, Dong TT, Tsim KW. Anti-oxidation activity of different types of natural Cordyceps sinensis and cultured Cordyceps mycelia. Phytomed 2001;8:207-212.