Determination of andrographolide content in self nano emulsifying drug delivery system (SNEDDS) for in vitro diffusion study using validated HPLC

Abstract

This research aims to validate the analytical methods of a rapid and straightforward HPLC-UV method for determining of andrographolide content in Self-Nano Emulsifying Drug Delivery System (SNEDDS) formulation during in vitro diffusion study. The assay used an XTerra® MS Cl8 column (l50 mm X 4.6 mm, five µm) with a mobile phase of methanol and water (70: 30), at 0.8 mL/min flow rate and UV detection of 229 nm. The study was prepared HCl pH 3.4, phosphate buffer pH 6.8 and saline buffer pH 7.4 as diffusion medium. The validation parameter was established involving the assess on linearity, precision, accuracy, LOD, and LOQ. The andrographolide SNEDDS diffusion test was performed using Franz diffusion cell with Capryol 90, tween 20 and PEG 400 as SNEDDS lipid-based. The result demonstrated good linearity with r = 0.999 in HCl pH 3.4 and phosphate buffer pH 6.8 medium, while r = 0.998 in saline buffer medium. Validation of the analytical method for the diffusion test demonstrated an accurate result with % recovery was found to be a range 99.14-102.39; 102.28-103.02; and 93.52%-104.85% in HCl pH 3.4, phosphate buffer pH 6.8 and saline buffer pH 7.4, respectively. The technique showed adequate precision, with a relative standard deviation (RSD) less than % Horwitz. LOD and LOQ were found 1.31 and 3.13 µg/mL in HCl pH 3.4 medium, 1.95 and 5.92 µg/mL in phosphate buffer medium, and 2.21 and 6.70 µg/mL. The diffusion study showed that the concentration of drug release 73.35, 59.53 and 42.03 µg in HCl pH 3.4, phosphate buffer pH 6.8 and saline buffer pH 7.4, respectively. In conclusion, the HPLC method developed appropriately for determination andrographolide content in SNEDDS formulation for the in-vitro diffusion study.