Abstract
BIO ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF LAPATINIB 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 Lapatinib in human plasma. Lapatinib is extracted from human plasma by Liquid Liquid Extraction by 50μl 1mM NaOH and 2.5mL of Ethyl Acetate and analyzed using a reversed phase isocratic elution on a Kromacil 100 C18 (4.6 X 50mM, 5μm) column. A acetonitrile: 5mM ammonium formate (80:20%v/v) pH adjusted to 3.80 using formic acid is used as mobile phase and detection was performed by Ultra triple quadrupole mass spectrometry LC-MS/MS using electrospray ionization in positive mode. Pioglitazone is used as the internal standard. The lower limit of quantification is 15.004ng/mL for lapatinib. The calibration curves are consistently accurate and precise over the concentration range of 15.004 to 2000.540 ng/mL in plasma for Lapatinib. This novel LC–MS/MS method shows satisfactory accuracy and precision and is sufficiently sensitive for the performance of pharmacokinetic, bioavailability and bioequivalence studies in humans.

Keywords: Lapatinib, LC-MS/MS, Bioanalytical Method Validation, Human Plasma, Liquid Liquid Extraction, Breast Cancer.


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