Simultaneous quantification of five bioactive phenylethanoid, iridoid, and flavonol glycosides in Duranta erecta L.: Ultra performance liquid chromatography method validation and uncertainty measurement.

Duranta erecta L.is a valuable herb used in traditional system of medicine in India and China for the treatment of malaria and abscesses. The presence of phenylethanoid (e.g., acteoside as major bioactive) in addition to iridoid and flavonoids glycosides are mainly responsible for reported pharmacological properties. Although D. erecta is widely used in many Asian countries, the quality assurance of raw herb and its derived products are still in question, due to the lack of validated protocol. Current Indian guidelines for quality assurance of phytopharmaceutical product demands accurate determination of minimum four chemicals. Moreover, there is no chromatographic method to simultaneously analyse acteoside, isoacteoside, durantoside I, quercetin, and methylapigenin-7-O-β-D-glucopyranuronate in D. erecta. The present study establishes a systematic approach for the quality evaluation of D. erecta with efficient and validated UPLC-PDA method that simultaneously analyses the five bioactive marker chemicals in D. erecta. A fast, sensitive, and reliable ultraperformance liquid chromatography-photodiode array (UPLC-PDA) method was developed for the quantification five marker compounds-acteoside, isoacteoside, durantoside-I, quercetin, and methylapigenin-7-O-β-D-glucopyranuronate in D. erecta. The optimized UPLC conditions on Reverse phase column Phenomenex Luna® (C₁₈, 2.5μm, 2.0×100mm)) and binary gradient elution resulted in best separation for all five targeted phytoconstituents. Validation characteristics viz. linearity (r² >0.999), accuracy (%RSD<2.0), precision (%RSD, 1.62-2.59), recovery (99-101%), and sensitivity (LOD- 0.27-0.40μg mL^-1; LOQ- 0.90-1.35μgmL^-1) were satisfying the ICH criteria. Uncertainty in the measurement was also estimated using method validation data and other sources to fulfil the technical requirement of ISO 17025:2017. It is the first validated method that provides the simultaneous and accurate analysis of five bioactive phytoconstituents of D. erecta in short time with defined traceability and accuracy profile. Further, it delivers a holistic quality analysis of raw medicinal herb and its preparation.