Quantification and comparison of extraction methods for alkaloids in Aegle marmelos leaves by HPLC

The leaves of Aegle marmelos are reported to contain multi-bioactive classes of compounds including coumarins, furanocoumarins and alkaloids. HPLC analysis of the crude extract was challenging due to low concentrations of the compounds in the leaves. Five compounds visible in the HPLC chromatogram were separated and identified by HPLC and further elaborated for quantification as marker compounds of A. marmelos leaves using a C18 column with detection at 275 nm. A gradient mobile phase consisting of acetonitrile and water was used. The developed HPLC method showed good linearity (r2 > 0.994), high precision (RSD<5%), and good recovery (99.27-99.98%) of the compounds. The lowest detection limit was 5 ng and the method was found to be robust. All the validation parameters were within the permissible limits. Therefore, the developed method is accurate and reliable for the quality control of A. marmelos. This is the first report of extensive quantitative HPLC analysis of marker compounds in A. marmelos leaves and method validation.