title: An extensive study on isosorbide-5-mononitrate acid adducts for quantification in human plasma using liquid chromatography/electrospray ionization tandem mass spectrometry and its application to bioequivalence sample analysis.
abstract: A rapid and sensitive liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) method for the determination of isosorbide-5-mononitrate (5-ISMN), used in the treatment of angina pectoris, in human plasma is described. The quantification of 5-ISMN was performed via stable acetate adduct formation with a high relative abundance. The plasma filtrate obtained after solid-phase extraction (SPE), using a polymer based, hydrophilic-lipophilic balanced (HLB) cartridge, was submitted directly to reversed-phase high-performance liquid chromatography separation followed by ESI and detection of the resulting ions using triple-quadrupole mass spectrometry in selected reaction monitoring (SRM) mode. There was no significant matrix effect on the analysis. For validation of the method, the recovery of the free analyte response was compared to that obtained from an optimized extraction method. The analyte stability was examined under conditions mimicking the sample storage, handling, and analytical procedures. The extraction procedure yielded extremely clean extracts with a recovery of 95.51% and 93.98% for iossorbide-5-mononitrate and topiramate (internal standard (IS)), respectively. The calibration curves were linear for the dynamic range of 10.0 to 1000.0 ng/mL with a correlation coefficient r > or = 0.9985. The intra-assay and inter-assay precision for the samples at the lower limit of quantification (LLOQ) were 9.02 and 13.30%, respectively. The intra-assay accuracies at LLOQ, LQC, MQC and HQC levels varied from 98.13 to 118.15, 102.34 to 105.21, 100.69 to 109.68, and 95.76 to 102.92%, respectively, while the inter-assay accuracies ranged from 93.10 to 118.15, 93.03 to 107.04, 86.97 to 109.68 and 86.18 to 105.85%, respectively, at these levels. The method is rugged and fast with a total run time of 2 min. The method was successfully applied for a bioequivalence study in 24 human subject samples after oral administration of 60 mg extended release (ER) formulations.
authors.affiliation: Department of Chemistry, School of Sciences, Gujarat University, Navrangpura, Ahmedabad 380 009, India.
authors.forename: Deepak S
authors.initials: DS
authors.lastname: Jain
authors.forename: Gunta
authors.initials: G
authors.lastname: Subbaiah
authors.forename: Mallika
authors.initials: M
authors.lastname: Sanyal
authors.forename: Pranav S
authors.initials: PS
authors.lastname: Shrivastav
authors.forename: Usha
authors.initials: U
authors.lastname: Pal
authors.forename: Shailesh
authors.initials: S
authors.lastname: Ghataliya
authors.forename: Abhijeet
authors.initials: A
authors.lastname: Kakad
authors.forename: Jignesh
authors.initials: J
authors.lastname: Bhatt
authors.forename: Vaneet
authors.initials: V
authors.lastname: Munjal
authors.forename: Heena
authors.initials: H
authors.lastname: Patel
authors.forename: Sapna
authors.initials: S
authors.lastname: Shah
mesh_terms: D000107:Acetylation; D000284:Administration, Oral; D002851:Chromatography, High Pressure Liquid; D003692:Delayed-Action Preparations; D006801:Humans; D007548:Isosorbide Dinitrate / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D020030:Nitric Oxide Donors / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013810:Therapeutic Equivalency
chemical_list: D003692:Delayed-Action Preparations; D020030:Nitric Oxide Donors; D007548:Isosorbide Dinitrate; C030397:isosorbide-5-mononitrate