Abstract

QUANTITATIVE ANALYSIS OF ANDROGRAPHOLIDE IN A STANDARDISED HERBAL EXTRACT USING THE UPLC TECHNIQUE

Buvana C.*, Naveenkumar R., Thenmozhi S., Rabiniraj S.*, Ragul V., S.M. Manjunath

Abstract

The objective of this study was to establish and authenticate a UPLC method for determining the level of Andrographolide in herbal formulation that include extract of Andrographics Paniculata (Burm f.). The Ultra Performance Liquid Chromatography (UPLC) technique was developed and subsequently validated to facilitate the efficient and uncomplicated determination of Andrographolide in different extracts. The experimental setup employed for this study consisted of a high-quality Nano UPLC Waters separation module, which was equipped with a PDA detector (190-700). The column employed for the experiment was an ACQUITY UPLC Bridge (C18 180 x 130mm, 1.7 - 3.5 μm). To make mobile phase A, 40 percent acetonitrile (ACN) and 60 percent phosphoric acid (0.1 percent) were mixed. Mobile phase B, on the other hand, was made by combining 70% methanol with 0% phosphoric acid (0.1%). Adjusting the flow rate to 0.5 ml/min and keeping the detecting wavelength at 230 nm required constant attention. The findings indicated that 0.15 ng/ml was the lower limit of detection (LOD) and that 0.27 ng/ml was the lower limit of quantitation (LOQ). A coefficient of determination (r2) of 0.987 indicated that the linearity of the calibration curve was in the range of 0.01-0.1 g/mL. The approach has a relative standard deviation (RSD) of 1.34% and an accuracy of 96.89% to 100.2%. We calculated an intra-day accuracy of 6.15% and an inter-day precision of 6.53%. The currently-being-developed herbal Nano emulsion is anticipated to be standardised using the validated approach.

Keywords: Andrographics Paniculata, validation, UPLC, Andrographolide,