Rapid Determination of Andrographolide and Dehydroandrographolide in <i>Andrographis paniculata</i> by HPLC-UV at Equal Absorption Wavelength

Abstract

A rapid and reference component saving method for the determination of andrographolide and dehydroandrographolide in Andrographis paniculata was developed in this study. The sample was prepared using matrix solid-phase dispersion and analyzed by an Agilent Poroshell 120 EC-C18 column (50 mm × 4.6 mm, 2.7 μm) with a water-acetonitrile mobile phase. The equal absorption wavelength of two components was confirmed to be 240 nm using a combination of ultraviolet spectrophotometer and high performance liquid chromatograph. Results showed that andrographolide and dehydroandrographolide exhibited good linearity in their concentration ranges (r ≥ 0.9999), with the limit of quantification of 80.85 μg/g and 80.63 μg/g, respectively. Two components also had a good precision as their relative standard deviation (RSD) value less than 1.00%. The average recoveries of two components were 102.80% (RSD = 1.54%) and 100.70% (RSD = 1.91%), respectively. The developed method shared the same determination result with external standard method and Chinese Pharmacopoeia, which took a total of 10 minutes, consumed 8.5 mL harmful solvent, and only used one reference component to determine the content of two components. This method is characterized by rapid determination and low reference component consumption in the rapid study on the main active components of Andrographis paniculata and its products, which contributes to the quality evaluation of Andrographis paniculata.