Validated Ultra Performance Chromatographic Method for Simultaneous Determination of Andrographolide and Its 4 Related Diterpenoids in Andrographis paniculata Capsules

A simple and rapid isocratic reverse phase chromatographic method was developed using UPLC for simultaneous determination of 5 diterpenoids; andrographolide, deoxyandrographiside, neoandrographolide, 14-deoxyandrographolide and 14-deoxy-11,12-didehydroandrographolide in Andrographis paniculata capsules. The chromatographic separation was achieved on a Waters Acquity BEH C18, 50 × 2.1 mm, 1.7 μm column using mobile phase consisting of acetonitrile and water in a ratio of 25:75 (v/v). The flow rate was 0.4 ml/min with a runtime of 12 min. The method was validated according to the regulatory guidelines with respect to specificity, linearity, precision, accuracy, limit of detection, limit of quantitation and robustness. The proposed method provided linear responses (r2>0.999) with good precision (%RSD<3%). The recovery values ranged from 98.00% to 101.36% for all compounds. The developed method was applied for analysis of 5 active compounds in capsule formulation.