Development and Validation of a Stability Indicating Method for Seven Novel Derivatives of Lamivudine With Anti-HIV and Anti-HBV Activity in Simulated Gastric and Intestinal Fluids.

GUALDESI, MARÍA S.; ESTEVE-ROMERO, JOSEP; BRIÑÓN, MARGARITA C., RAVIOLO, MÓNICA A.

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2013 vol. 78 p. 52 - 56

0731-7085

A simple micellar liquid chromatography (MLC) method has been developed and validate for use in stability indicating studies of lamivudine and its carbonate derivatives with proved activity against human immunodeficiency and hepatitis B viruses (HIV and HBV, respectively), in simulated gastric (SGF) and intestinal (SIF) fluids samples. The optimized method involve a C18 column thermostated at 30ºC, UV detection at 272 nm, a flow rate of 1.0 mL min-1 and a micellar mobile phase composed by 0.15 M sodium dodecyl sulphate (SDS) -4 % (v/v) 1-butanol-0.01 M KH2PO4-Na2HPO4 (pH 7), using zidovudine (AZT) as internal standard. Validation under Food and Drug Administration (FDA) guideline of the analytical parameters include: linearity (r2 0.9996), LODs (1.6x10-7-6.9x10-6 M) and LOQ (1x10-5 M), intra (0.02-1.48%) and inter-day precision (0.04-1.66%) expressed as relative standard deviation (RSD), and robustness (less than 1.98%). Using this method, recoveries ranging from 92.9 to 119% were obtained for the eight substances. Thus, this method provides a simple, sensitive, precise and reproducible assay for all the compounds that can be readily adaptable to routine use by clinical laboratories with standard equipment. In addition, we evaluated the stability of carbonates of lamivudine in buffer pH 1.2 and 6.8; SGF (pH 1.2) and SIF one (pH 6.8), all as indicated in United States Pharmacopeia (USP) 32. Finally, this MLC method was applied to stability studies which resulted in all the compounds following a pseudo-first-order kinetics, and in the determination of its kinetic constant and half-life time.