Screening the elite chemotypes of Gloriosa superba L. in India for the production of anticancer colchicine: simultaneous microwave-assisted extraction and HPTLC studies

Abstract

Background Gloriosa superba L. (Colchicaceae) is a high-value medicinal plant indigenous to Africa and Southeast Asia. Its therapeutic benefits are well-established in traditional medicines including Ayurveda. It is well known for its natural bioactive compound colchicine which exhibits a wide range of pharmacological activities i.e. rheumatism, gout and was also introduced into clinical practices. The increasing demand as well as its illegal harvesting has brought this valuable plant under threatened category. Methods The present investigation describes a microwave assisted extraction (MAE) strategy coupled with a densitometric-high performance thin layer chromatographic (HPTLC) methodology for the analysis of colchicine from 32 different populations of G. superba. A Box-Behnken statistical design (3 level factor) has been employed to optimize MAE, in which power of microwave, time of irradiation, aqueous ethanol and pH were used as independent variables whereas colchicine was used as the dependent variables. Chromatography was carried out on Silica gel 60 F254 TLC plates with toluene: methanol, 85:15 (v/v) being used as solvent system. Densitometric measurement was performed at λ=254 nm following post-derivatization (10% methanolic sulphuric acid). Results Optimal conditions for extraction to obtain the maximum colchicine yield was found to be 7.51 mg g− 1 which was very close to be predicted response 7.48 mg g− 1 by maintaining microwave power (460 W), irradiation time (6.4 min), aqueous ethanol-30, pH -3. Colchicine content ranged between 2.12–7.58 mg g− 1 among 32 G. superba populations in which only three chemotypes viz. GS- 1, GS- 3, and GS- 2 collected from West Bengal and Sikkim, respectively exhibited maximum yield of colchicine. Conclusion Therefore, this newly developed optimized MAE coupled with HPTLC densitometry methodology not only quantifies colchicine in order to identify elite chemotypes of G. superba, but it also encourages in selecting high yielding populations of the plants for industrial use and economic boost for the farmers. This validated, simple and reproducible HPTLC protocol is being used for the first time to estimate colchicine from natural populations of G. superba obtained from 32 different geographical regions of India. Graphical abstract

Determination of Colchicine in Pharmaceutical Formulations, Traditional Extracts, and Ultrasonication-Based Extracts of Colchicum autumnale Pleniflorum (L.) Using Regular and Greener HPTLC Approaches: A Comparative Evaluation of Validation Parameters

“…For its determination in serum and plasma samples, various liquid chromatography-mass spectrometry (LC-MS) methods have been reported [1,[22][23][24]. For the determination of this compound in various commercial formulations and plant extracts, distinct high-performance thin-layer chromatography (HPTLC) assays have been reported [10,[25][26][27][28][29][30][31][32][33][34][35]. Some electrochemical techniques have also been utilized for the determination of this compound [36,37].…”

Vegetative anatomy and endorrhizal fungal morphology of an endangered medicinal plant Gloriosa superba L.

“…Being a mycorrhizal plant, association with arbuscular mycorrhizal (AM) fungi would aid G. superba in its growth and secondary metabolite production. For instance, Pandey et al (2021) showed that inoculation of AM fungi along with or without phosphorous fertilizer supplementation increased colchicine content in the tubers of G. superba . In addition to AM fungi, the roots of G. superba are also colonized by the ascomycetous dark septate endophytic (DSE) fungi (Muthukumar et al, 2006).…”

“…It is categorized as endangered by International Union for Conservation of Nature (IUCN) in India due to overexploitation of the species for various medicinal purposes (Tilwari & Sharma, 2021; Yadav et al, 2012). Colchicine is one of the important alkaloids present in G. superba that helps in treating various human ailments (Balkrishna et al, 2019; Pandey et al, 2021). Therefore, it is important to conserve this plant in its natural habitats for the future.…”

“…In India, it is distributed throughout the tropical regions ranging from the North‐West Himalayas to the Deccan peninsula and Western Ghats of Karnataka, Kerala, and Tamil Nadu (Maroyi, 2013; Patil & Gavade, 2012). In addition, G. superba is the national flower of Zimbabwe as well as the state flower of Tamil Nadu in India (Ashokkumar, 2015; Pandey et al, 2021).…”

Medicinal pteridophytes: ethnopharmacological, phytochemical, and clinical attributes

“…As plants proceed through their normal metabolic processes, they form phytochemicals. The term "secondary metabolites" refers to the many kinds of chemicals which can be found in plants, namely alkaloids, coumarins, flavonoids, glycosides, gums, phenols, polysaccharides, tannins, terpenes, and terpenoids [80,126]. Plants also contain other chemical compounds in addition to these.…”