Employment of ATR-FTIR and HPLC-UV method for detection and quantification of andrographolide

Abstract

Objective: The purpose of this present study was to describe the employment of infrared (IR) spectroscopy and high-performance liquid chromatography with ultraviolet detection (HPLC-UV) method in determining the identity and purity of bulk material containing andrographolide. Methods: Attenuated total reflectance (ATR)-Fourier transform infrared (FTIR) spectroscopy was carried out in transmittance mode to investigate the molecular vibration of the bulk material component and the similarity of functional groups between bulk material and standard andrographolide. Meanwhile, the liquid chromatographic analyses were performed on a reversed-phase C18 column and under UV detection at 224 nm to determine the andrographolide content of the bulk material. Results: The obtained ATR-FTIR spectra indicated that functional group of the bulk material were in close similarity with those of standard andrographolide. The linearity of the evaluated method achieved over a concentration range of 1 to 60 μg/ml with a high correlation coefficient (0.999). By using the studied HPLC-UV method, the andrographolide of bulk material was found to be 98.12% (retention time of 2.58 min). Conclusion: The studied HPLC-UV method of andrographolide determination is accurate, precise, selective, and brief in terms of analysis time. The studied method, therefore, provides a rapid and reliable assessment for identifying and determining the purity of andrographolide bulk material.