Analytical Method Development, Validation, and Estimation of Lupeol, Quercetin, Vasicine in Polyherbal Formulations and Selected Plant Species by using UFLC-MS

Abstract

Background: Kabusura kudineer (KSK) is a marketed Siddha-based polyherbal formulation, and on the authority of the Govt of India, the Ministry of Ayush recommended Kabusura kudineer in the therapy of COVID-19, and due to its immuno-booster effects. Based on the “Kabusura kudineer” (KSK) formulations, we developed a “Hydaljss08” polyherbal formulation composed of 16 dried crude materials and 03 fresh forms. Both formulations contain active phytopharmaceuticals such as vasicine, quercetin, and lupeol and these are responsible for anti-viral and immunomodulatory effects, which may be due to their synergistic and additive effects. The chemical nature of vasicine is an alkaloid, quercetin is a flavonoid, and lupeol is a pentacyclic triterpenoid. Aim: The current study aims to develop and validate the analytical process for assessing vasicine, quercetin, and lupeol in both dosage forms and in a selected plant species by UFLC-MS. Methods: The ultrafast liquid chromatography study was designed by the columns of Inertsil C8, and Inertsil C18, individually, for vasicine, quercetin, and lupeol respectively. The columns and mobile phase were used as a water C18, 20 mM phosphate buffer pH 2.5: acetonitrile in a combined UFLC method development of vasicine, quercetin, and lupeol. Results: A calibration curve and adequate linearity were recorded for vasicine, quercetin, and lupeol by injecting 20.0-60.0 μg/ml, 50.0-150.0 μg/ml, and 25.0-75.0 μg/ml of marker substances. The LOD, and LOQ of the vasicine, quercetin, and lupeol were found to be 1.19, 3.60, 3.80, 11.51, 1.79, and 5.41 μg/ml, and the Ruggedness value of vasicine, lupeol, and quercetin was found in % RSD 0.4%, 0.1%, and 0.1%. Conclusion: The developed and validated method showed good linearity with a range of correlation coefficients, new, simple, novel, accurate, specificity, precision, robustness, and ruggedness are within the limits.