The current study focused on the optimization of a rapid in-vitro-culturing protocol and analytical characterization of the Cordyceps militaris (L.) Link fruiting bodies. The main intension of the study is to increase the yield and bioactive metabolite content in the Cordyceps militaris. The tissue engineering process, the solid-state fermentation technique, used in the culturing of the Cordyceps militaris (L.) Link. The different cultural conditions were optimized to increase the growth and metabolite production. The main parameters were optimized with respect to physicochemical condition and to growing media. The temperature, pH, and shaking speed were systematically optimized to maximize growth and production of metabolite the temperature at 19–21°C, pH 5.5–6.0, and 120–150 rpm established the uniform and good growth of the mycelium. The Potato Dextrose Broth (PDB) confirmed the uniform growth and yield of the Cordyceps militaris fruiting bodies. The nutrient broth supplemented with brown rice used as substrate in the process of mycelial growth and taken 50–55 days, achieving a biological efficiency of 90–110%.
The phytochemical screening of the cordyceps militaries shows the presence of major bioactive components, including phenolics, flavonoids, saponins, and proteins. The UV- spectroscopic analysis revealed the assay of total phenolic and flavonoid contents of 14.36% and 0.81% (w/w), respectively. The sequential solvent extraction demonstrated maximum recovery with ethyl acetate and hydroalcoholic solvents. The HPLC findings identified adenosine (1.20%) and cordycepin (3.73%) with retention times of 4.5 and 5.3 minutes, respectively, indicating efficient extraction of pharmacologically relevant nucleosides. The cordyceps militaries extract was subjected to GC–MS and LC-MS analysis and GC-MS revealed 80 volatile and semi-volatile compounds, including esters, fatty acids, and nitrogenous derivatives. The LC–MS confirmed the presence of inosine and 5-methyluridine as key polar metabolites. The study confirms the in-vitro culture of Cordyceps militaris can serve as a reproducible and sustainable system for producing high-quality fruiting bodies rich in therapeutic and nutritional constituents comparable to previous cultivation studies.
Keywords: Cordyceps militaris, solid-state fermentation, optimization, cordycepin, adenosine, HPLC, GC–MS, LC–MS, phytochemical profiling, nutraceutical potential.
How to cite this article: Sukanya P, Chandan S, Shivaprasad K Chandan D and Pradeep S; In-Vitro Cultivation Optimization of Cordyceps militaris and Analytical profiling: Implications for Nutraceutical and Functional Food Development. Int J Drug Deliv Technol. 2026;16(1): 321-336. DOI: 10.25258/ijddt.16.1.35