Simultaneous Quantification of Major Bio‐Active Diterpenoid Lactones and Flavonoids in Andrographis paniculata (Burm. F.) Nees: LC‐ESI‐MS/MS Method Validation and Uncertainty Determination

Abstract

AbstractDiterpenoid lactones and flavonoids are major bio‐active compounds of therapeutically important medicinal herb Andrographis paniculata (Burm.F.) Nees. The lack of a validated protocol for quality assurance of raw herbs is a significant issue for its wider use. A validated LC‐ESI‐MS/MS method developed using MRM mode for simultaneous determination of five diterpenoid lactones (andrographolide, neoandrographolide, andrograpanin, 14‐deoxy‐11,12‐didehydroandrographolide and andrographiside) along with two flavonoids (7‐O‐methylwogonin and apigenin). Chromatographic separation of seven analytes was achieved within 14 minutes in Alltima column (100×4.6 mm, 3 μm) using gradient elution. The developed method was accurate (97.77 to 101.17 %; recovery), precise (intra‐day and inter‐day %RSD of 0.22–2.19 and 1.01–3.68, respectively) and linear (R2 >0.99). To meet the regulatory obligation, uncertainty associated with the measurement was also evaluated by using different validation parameters. Developed method was successfully employed in analysing the analytes quality in different solvent extracts. Total diterpenoid lactones and flavonoids (DTLF) content was recorded highest in methanol extract (45.47 μg g−1) followed by ethanol (39.23 μg g−1) and lowermost content found in petroleum ether extract (0.38 μg g−1) in triplicate samples. Since the method is simple, rapid, sensitive, and accurate, it has potential role for quality evaluation of A. paniculata and its derived formulation.