HPLC-Based Elucidation of Tannins from the Tissue and Callus Culture Extracts of Selected Medicinal Plants
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Background. Optimized HPLC profiling is a powerful and effective analytical tool to standardize plant samples and authenticate plant materials. In this study, three selected medicinal plants namely: Achyranthes aspera, Ipomoea hederacea, and Ocimum basilicum were subjected to callus induction following seedling, leaf, and stem germination.
Methods. 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. HPLC fingerprinting was performed using a Shimadzu LC-20A system with a retention time of 2.9 minutes at 270 nm. The aim was to ensure quality and consistency in tannin analysis across different plant parts and callus culture samples.
Results. The highest callogenic response occurred in A. aspera leaf explants on MS medium with 2.0 mg/L 2,4-D and 4.0 mg/L NAA, producing green, granular callus. The lowest was in I. hederacea stem explants with 0.5 mg/L 2,4-D and BAP, yielding brown, granular callus. O. basilicum leaf callus extract showed the largest sample area (9365.56) and tannin content (2.66), with superior precision in tannin analysis for O. basilicum and A. aspera (7.81).
Conclusion. HPLC profiling proved to be an accurate, efficient, and precise method for evaluating tannin content in selected plant samples. It is a crucial method to standardize the quality of medicinal plant compounds.
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