AN HPLC METHOD FOR THE ESTIMATION OF ANDROGRAPHOLIDE IN RABBIT SERUM

Objective: To develop a HPLC method for the estimation of andrographolide in biological fluids. Methods: Andrographolide was extracted from serum using methanol. A Shimadzu HPLC (Model LC 10 A) with UV- Vis spectrophotometric detector (model SPD 10 AV) and a data processor (model 10 R4 A) were used for andrographolide estimation. A Neuclosil C 18 octadecyl silane (ODS) column (5μ, 250x4.6 mm Hypersil) was used. The mobile phase used was methanol: Water in the ratio of 65:35 v/v at a flow rate of 1.0 ml/min. The elutes were monitored at 223 nm. The precision and accuracy of the method were determined by measuring inter-day and intra-day variations. Results: The absolute recovery of andrographolide ranged between 97.24% to 99.4%. The inter-day co-efficient of variation was 0.4% to 1.4% and intra-day co-efficient of variation was 1.05% to 2.8%. Conclusion: The developed HPLC method for estimation of andrographolide in serum is sensitive, reliable and accurate.