HPLC-Based Elucidation of Tannins from the Tissue and Callus Culture Extracts of Selected Medicinal Plants
Optimization and Analytical Profiling for Sustainable Tannin Extraction
Abstract
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Optimized HPLC profiling is a powerful and effective analytical tool for standardizing plant samples and authenticating plant materials. In this study, three selected medicinal plants: Achyranthes aspera, Ipomoea hederacea, and Ocimum basilicum were subjected to callus induction following seedling, leaf, and stem germination. The induced callus was subsequently dried, finely ground, and extracted using methanol and water for HPLC analysis. A validated procedure was employed to identify and separate the tannin content in seedling leaf, stem, and callus culture extracts. The highest callogenic response was observed in A. aspera leaf explants cultured on MS medium supplemented with 2.0 mg/L 2,4-D and 4.0 mg/L NAA. These callus cultures exhibited a green color and granular morphology. Conversely, the lowest callogenesis response was noted in I. hederacea stem explants grown on MS medium supplemented with 0.5 mg/L 2,4-D and 0.5 mg/L BAP, resulting in brown-colored, granular callus.
HPLC fingerprinting was performed using a Shimadzu LC-20A system with a retention time of 2.9 minutes at 270 nm to ensure quality consistency in tannin analysis across different plant parts. Among all explants and callus culture samples, O. basilicum leaf callus culture extract exhibited the highest sample area (9365.56) and the highest tannin content (95.04%). Furthermore, the tannin content of O. basilicum (2.66) and A. aspera (7.81) leaf callus culture extracts demonstrated superior precision and accuracy in relative standard deviation values compared to stem seedling tissues and other callus culture extracts.HPLC profiling proves to be an accurate, efficient, and precise method for evaluating tannin content in selected plant samples. Moreover, these plants hold significant potential for isolating bioactive compounds in pure form, which could be harnessed for pharmaceutical applications. The optimized chromatographic fingerprint serves not only as an alternative analytical tool for authentication but also as a crucial method for standardizing the quality of medicinal plant compounds.
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