Development and Validation of Highly Sensitive HPLC-Ms/Ms Method for the Determination of Duloxetine in Human Plasma and its Application to Clinical Pharmacokinetic Study by Assessing Multiple Bioequivalence Approaches
*Corresponding Author E-mail: email@example.com
Duloxetine (DUX) is a potent selective serotonin and norepinephrine reuptake inhibitor used to treat depression and anxiety in human. The concentration of DUX in biological matrix is highly variable; hence it is necessary to have a highly sensitive and selective analytical method to measure the anticipated range in the plasma. Thus, there is a necessity to develop a sensitive and practical analytical method for the determination of DUX in human plasma.
The objective of the study is to estimate DUX in human plasma using high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) and its application to clinical pharmacokinetic study by assessing multiple bioequivalence approaches.
The analyte was extracted by simple liquid-liquid extraction technique using n-hexane as an extraction solvent. The chromatographic separation was carried out by using Luna® 5 μm C8 (2) 100 Å, LC Column 100 x 4.6 mm with the mobile phase consisting of Milli q water (0.05% formic acid and 0.1% ammonium trifluroacetate solution) and Methanol (23: 77%v/v) at 0.5mL/min flow rate. The MRM transitions of 298.00 → 154.00 and 256.00 → 148.00 were used for the quantification of DUX and Atomoxetine (internal standard) separately.
The method was validated over the concentration range of 0.100–100.000 ng/mL, the coefficient of determination was ≽ 0.9967. The sensitivity of the method was 0.100ng/mL with the accuracy and precision of 108% and 5.12% respectively. All validation parameters are found to be within the acceptable limits as per the USFDA bioanalytical method validation guidelines.
We have developed and validated a highly sensitive analytical method for the determination of DUX in human plasma using high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The developed method was successfully applied for analyzing the samples of clinical pharmacokinetic study conducted in healthy human adult subjects and the pharmacokinetic data was analyzed by using average, population and individual bioequivalence approaches. The method was very rugged and is capable of continuously analyzing the clinical pharmacokinetics study samples without any run failures.
Bioanalytical Method, Duloxetine, Average Bioequivalence, Individual Bioequivalence, Population Bioequivalence.