European Journal of Biomedical and Pharmaceutical Sciences

BIO ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF AMODIAQUINE AND N-DESETHYL AMODIAQUINE IN HUMAN PLASMA BY LC–MS/MS.

Premanand Ranganathan*, V. Gunasekaran and Indrajeet Singhvi

ABSTRACT

A highly sensitive and simple high performance liquid chromatographic tandem mass spectrometric (LC–MS/MS) assay is developed and validated for the quantification of Amodiaquine and N-Desethyl Amodiaquine in human plasma. Amodiaquine and N-Desethyl Amodiaquine is extracted from human plasma by methyl tertiary butyl ether and analyzed using a reversed phase isocratic elution on a Hypersil Gold (4.6 x 50mm, 4.6μm) column. A acetonitrile: 10mM Ammonium formate with 0.1% Trifluoro acetic acid (75:25 v/v) as mobile phase is used and detection was performed by Ultra triple quadrupole mass spectrometry LC-MS/MS using electrospray ionization in positive mode. Amodiaquine D10 and N- Desethyl Amodiaquine D5 are used as the internal standards. The lower limits of quantification are 0.106 ng/mL and 0.523ng/mL for Amodiaquine and N-Desethyl Amodiaquine respectively. The calibration curves are linear over the concentration range of 0.106 to 51.580 ng/mL for Amodiaquine and 0.523 to 253.855 ng/mL for N-Desethyl Amodiaquine of plasma for each analyte. This novel LC–MS/MS method show satisfactory accuracy and precision and is sufficiently sensitive for the performance of pharmacokinetic studies, Bioavailability studies and Bio Equivalence Studies in humans.

Keywords: Amodiaquine, N-Desethyl Amodiaquine, LC-MS/MS, Human Plasma, Method Validation.