ABSTRACT
Obesity can be considered as a chronic illness of epidemic proportion and its incidents have increased exponentially in recent years.The use of anti-obesity drugs such as sibutramine is somewhat helpful.There is a need to quantify such drugs in biological samples,which is generally quite difficult.In this report,we developed and validated a simple,sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of sibutramine (SB) and its two metabolites N-des methyl sibutramine (DSB) and N-di desmethyl sibutramine (DDSB) in human plasma.Zorbax SBC18 (4.6 mm × 75 mm,3.5 μm,80 (A)) analytical column and 5 mM ammonium formate:acetonitrile (10∶90,v/v) mobile phase were used for chromatographic separation of SB,DSB and DDSB.Multiple reaction monitoring (MRM) in the positive mode was used to detect SB,DSB and DDSB at m/z 280.3/124.9,266.3/125.3 and 252.2/124.9,respectively.Liquid liquid extraction was used for the extraction of analytes and internal standard from human plasma.This method was validated over a linear concentration range of 10.0-10,000.0 pg/mL for SB,DSB and DDSB with correlation coefficients (r) of ≥0.9997.The drug and the two metabolites were stable in plasma samples.The validated method was successfully applied in a bioequivalence and pharmacokinetic study with human volunteers under fasting condition.
ABSTRACT
A simple,sensitive,and specific liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of desloratadine (DL) in human plasma using desloratadine-d5 (DLD5) as an internal standard (IS).Chromatographic separation was performed using an Xbridge C18 column (50 mm × 4.6 mm,5 μm) with an isocratic mobile phase composed of 10 mM ammonium formate:methanol (20∶80,v/v),at a flow rate of 0.7 mL/min.DL and DLD5 were detected with proton adducts at m/z 311.2→259.2 and 316.2→264.3 in multiple reaction monitoring (MRM)positive modes,respectively.Liquid-liquid extraction (LLE) method was used to extract the drug and the IS.The method was validated over a linear concentration range of 5.0-5000.0 pg/mL with a correlation coefficient of (r2)(≥)0.9994.This method demonstrated intra- and inter-day precision within 0.7-2.0％ and 0.7-2.7％,and an accuracy within 101.4-102.4％,and 99.5-104.8％.DL was found to be stable throughout the freeze-thaw cycles,bench-top,and postoperative stability studies.This method was successfully applied in the analysis of plasma samples following oral administration of DL (5 mg) in 35healthy Indian male human volunteers under fasting conditions.