Novel “HYDALJSS08” Hydroalcoholic Polyherbal Formulation Development and Ultra-performance Liquid Chromatographic Separation, Estimation of Andrographolides in Andrographis paniculata whole Plants and a Marketed Siddha-based Polyherbal Formulation “Kabusura Kudineer”

Abstract

Background: The Siddha-based polyherbal formulation known as “Kabusura Kudineer (Marketed)" and developed as “HYDALJSS08” hydroalcoholic polyherbal formulation contains some fifteen plant materials in a dried raw form. Due to its immuno-booster properties, the Ministry of Ayush, Govt of India, highly recommended the use of "Kabusura Kudineer" during the pandemic of COVID-19. Objective: The present study intends to expand and validate the analytical profile for Andrographolides (AP), and isolated Andrographolides (AP) from the Andrographis Paniculata whole plant and in the Polyherbal Formulations (Marketed-Kabusura Kudineer, & Developed “HYDALJSS08”). Methods: One of the active components of “Kabusura Kudineer” marketed and developed as “HYDALJSS08” Hydroalcoholic Polyherbal formulation is kalmegh, also known as the king of bitter (Andrographis Paniculata-Acanthaceae). Kalmegh composes active principal components of Andrographolides (AP), which are proven for their Anti-viral and immunomodulatory activity. The preliminary identification of AP and the sample was carried out by TLC and FT-IR. The liquid chromatography was performed on a Zorbaz SB C8 (250*4.6mm & 5μm). The mobile phase incorporated pH 2.8 phosphate buffer with Acetonitrile: Methanol (60:30:10). The flow rate of the mobile phase was 1ml/min, and effluents were kept an eye on at 223 nm in a UV detector. The run time on the chromatogram was 10 min, and retention time was also observed. Results: The Rf value of Andrographolides (AP) was found to be 0.62. ICH guidelines were followed to carry out the Validation parameter. The retention time of AP was 2.5 min, and the Valid parameters of AP and system precision were as follows: SD (1831.11), % RSD (0.2), regression equations y = 41978 + x−10763, and correlation coefficient (R2) 0.9994. The adequate Linearity concentration was found to be 5 to 50 μg/ml, the value of LODs was 0.61μg /ml, LOQs was 2.01 μg/ml, method precision % RSD was 0.2, SD was 1597.1, and recovery was 99.9% and 101%. AP content found in a formulation (“Kabusura Kudineer” 1.48 μg/mL, developed “HYDALJSS08” Hydroalcoholic Polyherbal formulation-0.48 μg/ml) and isolated Andrographolides from Andrographis paniculata was 112.4μg/ml. Conclusion: The developed HPLC methods enabled simple, novel, rapid, easy, accurate, reproducible, and linear analysis of isolated andrographolides, and Siddha-based Polyherbal formulations.