Development and Validation of a Simple HPLC-UV Method for The Quantification of Andrographolide In Rabbit Plasma

Abstract

In this present work, a simple, rapid and accurate HPLC-UV method has been developed for the quantification of andrographolide in rabbit plasma. The assay was performed using an XTerra® MS C18 column (150 mm X 4.6 mm, 5 μm) with a mobile phase of methanol and water (60:40), at 0.8 mL/min flow rate and UV detection of 229 nm. Andrographolide was extracted from a biological sample by applying acetonitrile as a precipitating and extraction solvent. The results showed a good linearity with r = 0.9992; the accuracy reported as % diff was found to be -6.42 – 6.55 % while the recovery was 99.09, 98.55, and 105.14% for low, medium and high spiked plasma, respectively. The precision (reproducibility) reached 1.08–3.20 % RSD for the sample studied. The 2.87 % relative standard deviation (RSD) value for selectivity test indicated a good selectivity of the developed method. The developed method is simple and rapid, so that it can be applied for the quantification andrographolide in animal models during pharmacokinetics studies.