Recent Advancements of High-Performance Thin Layer Chromatography Method for Identification and Quantification of Oleanolic Acid in the Roots of Achyranthes aspera Linn.

Aim: A correlation exists between biological activities and presence of secondary metabolites in
plants. The present research work is carried out to develop and validate a chromatographic method
using HPTLC for identification and quantification of Oleanolic Acid from the roots of Achyranthes
aspera Linn.
Methods: The External Standard Method is used for quantification analysis in present study. It
assures accuracy and precision in quantitative analysis and is especially suited for HPTLC studies.
Chromatography was performed on aluminium-backed silica gel 60F254 HPTLC plates of 250 μm
thickness with solvents toluene: ethyl acetate: Methanol: Acetone 14:4:1:1 (v/v/v/v) as the mobile
phase. Derivatisation was carried out with Anisaldehyde sulphuric acid, scanned, and quantified at
540 nm.
Results: The concentration of Oleanolic Acid in roots of the plant is found to be 1.277ng/ μg. The
statistical analysis proved that the developed method is suitable and specific.
Conclusion: The developed HPTLC technique can be used for the routine quality control analysis
and quantitative determination of oleanolic acid from A. aspera Linn. The oleanolic acid was found to
be linear in the range of 0.20-0.60 μg/μl. Considering the wide therapeutic applications of oleanolic
acid an alternative quantification technique of this marker constituent was generated to ensure identity
and quality of the selected plant. This is a sensitive, specific and reproducible HPTLC method for the
quantification of Oleanolic Acid from roots of Achyranthes aspera Linn.