title: Development and fit-for-purpose validation of a LC-MS/MS assay for fibrinogen peptide A quantitation in human plasma.
abstract: BACKGROUND
  Fibrinopeptide A (FPA) is a plasma peptide, formed by the action of thrombin on fibrinogen during clog formation. FPA represents a direct indicator of thrombin activity and could potentially be used as a biomarker for anti-thrombotic therapy development. Results/Methodology: A LC-MS/MS assay with a high throughput solid phase extraction procedure was developed and validated to measure FPA in plasma. The lower limit-of-quantitation (LLOQ) of this assay was determined to be 0.16 nM. The inter- and intra-day%CV was <15%. Freeze-thaw stability of FPA was ±30% up to 3 cycles and linear response of FPA was observed for plasma dilution up to 16-fold.
  
  
  CONCLUSIONS
  The assay was validated and the biological variability of FPA in plasma was established (1-30 nM).
authors.affiliation: Clinical Development Laboratory, Merck Research Laboratories, Rahway, NJ, USA.
authors.forename: Mingxiang
authors.initials: M
authors.lastname: Lin
authors.forename: Michael E
authors.initials: ME
authors.lastname: Lassman
authors.forename: Russell
authors.initials: R
authors.lastname: Weiner
authors.forename: Omar F
authors.initials: OF
authors.lastname: Laterza
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D005344:Fibrinopeptide A / Q000032:analysis* / Q000302:isolation & purification / Q000592:standards; D005615:Freezing; D006801:Humans; D008969:Molecular Sequence Data; D055550:Protein Stability; D012015:Reference Standards; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000592:standards
chemical_list: D005344:Fibrinopeptide A