Abstract
A selective, high sensitive and high throughput liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS) method has been developed and validated for the chromatographic separation and quantitation of duloxetine in human EDTA plasma using fluoxetine (IS) as an internal standard. Analyte and IS were extracted from human plasma by liquid-liquid extraction using MTBE-n Hexane (80:20).The eluted samples were chromatographed on X-terra RP8 (50 mmx4.6 mm, 5 μm particle size) column by using mixture of 30 mM ammonium formate (pH-5.0±0.05) and acetonitrile as an isocratic mobile phase at a flow rate of 0.40 mL/min and analyzed by mass spectrometer in the multiple reaction monitoring (MRM) using the respective m/z 298.08→154.0 for duloxetine and 310.02→148.07 for IS. The linearity of the response/ concentration curve was established in human plasma over the concentration range 0.100-100.017 ng/mL. The lower detection limit (LOD,S/N>3) was 0.04 ng/mL and the lower limit of quantization (LOQ,S/N>10) was 0.100 ng/mL. This LC-MS/MS method was validated with Intra-batch and Inter-batch precision of 5.21-7.02. The Intra-batch and Inter-batch accuracy was 97.14-103.50 respectively. Recovery of duloxetine in human plasma is 80.31% and ISTD recovery is 81.09%. The main pharmacokinetic parameters were Tmax(hr) = (7.25±1.581), Cmax(ng/mL) (44.594±18.599), AUC0→t, = (984.702±526.502) and AUC0→∞, (1027.147±572.790) respectively.
Highlights
Duloxetine HCl (DLX) (Figure 1) is chemically, 2(+)-(S)-N-methyl-(gamma)-(1-naphthyloxy)-2 thiophenepropylamine hydrochloride[1]
The empirical formula is C18H19NOS.HCl and having a molecular weight of 333.88. It is used for the treatment of naturopathic pain associated with peripheral neuropathy especially diabetic polyneuropathy for which it is first-line and as an add-on treatment in stress urinary incontinence instead of surgery[4,5], indicated for the management of fibromyalgia[6,7]
The assessment of relative matrix effect was based on direct comparison of the MS/MS responses of the analytes spiked into extracts originating from different lots of plasma
Summary
Duloxetine HCl (DLX) (Figure 1) is chemically, 2(+)-(S)-N-methyl-(gamma)-(1-naphthyloxy)-2 thiophenepropylamine hydrochloride[1]. The empirical formula is C18H19NOS.HCl and having a molecular weight of 333.88 It is used for the treatment of naturopathic pain associated with peripheral neuropathy especially diabetic polyneuropathy for which it is first-line and as an add-on treatment in stress urinary incontinence instead of surgery[4,5], indicated for the management of fibromyalgia[6,7]. The standard stock solution of 1 mg/mL of duloxetine and fluoxetine was prepared by dissolving requisite amount in methanol. Calibration standards and quality control (QC) samples were prepared by spiking (2% total volume of blank plasma) blank plasma with stock solution. To an aliquot of 300 μL of spiked plasma sample, 50 μL internal standard was added and vortexed for 20 s. The dried samples were reconstituted with 300 μL of mobile phase and 20 μL was injection in the chromatographic system
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