Stability Indicating Bioanalytical Validation of Elexacaftor, Ivacaftor and Tezacaftor using HPLC in Human Plasma

Abstract

A simple, precised, accurate method was developed for the estimation of Elexacaftor, Ivacaftor and Tezacaftor in human plasma using the Lumacaftor as internal standard by RP-HPLC (Reverse phase-High performance Liquid Chromatographic) technique. Chromatographic conditions used are stationary phase Azilent (250 x 4.6mm, 5m), Mobile phase 0.01N Potassium di-hydrogen phosphate (pH: 3.5) : Acetonitrile in the ratio of 70:30(v/v) and flow rate was maintained at 1.0ml/min, detection wave length was 250nm, column temperature was set to 30oC and diluent was mobile phase Conditions were finalized as optimized method. Retention time of Ivacaftor, Elexacaftor and Tezacaftor were found to be 2.391min, 3.208min and 3.644min. %CV of the Elexacaftor, Ivacaftor and Tezacaftor was found to be 0.08%, 1.05% and 3.59%. %Recovery was obtained as 96.41%, 95.029% and 98.21%. The linearity concentration is in the range of 435-17400ng/mL of Elexacaftor, 60-2400ng/mL of Ivacaftor and 300-1200ng/mL of Tezacaftor (r2 = 0.999) .The lower limits of quantification were 435ng/mL of Elexacaftor, 600ng/mL of Ivacaftor and 300ng/mL of Tezacaftor which reach the level of both drugs possibly found in human plasma. Further, the reported method was validated as per the ICH guidelines and found to be well within the acceptable range. The proposed method is simple, rapid, accurate, precise, and appropriate for pharmacokinetic and therapeutic drug monitoring in the clinical laboratories.