title: Validation of an automated chromogenic assay of potency of factor VIII in commercial concentrates.
abstract: The determination of factor VIII (FVIII) potency in FVIII concentrates can be performed using both manual and automated methods. This work aimed to validate the use of the chromogenic kit Coamatic® FVIII (Chromogenix) on the automated ACL® Elite PRO analyzer for evaluating the potency of FVIII in commercial preparations in pharmaceutical analytical laboratories. After setting the activation and reading times to 2 min and 3 min, respectively, the validation parameters, according to the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guideline Q2 (R1), were as follows: linearity, expressed by the adjusted model: log (Absorbance) = 1.848 + 0.777∙log (Concentration), with r² = 0.998; accuracy was verified (P-value = 0.6959); and the coefficient of variation for repeatability and intermediate precision was ≤6.5%. The Coamatic® FVIII kit method has been adapted to the ACL Elite PRO analyzer with improved performance compared with a manual microplate method.
authors.affiliation: Biological Products and Biotechnology Division, Spanish Medicines Agency, Madrid, Spain. mvcollazo@aemps.es
authors.forename: V
authors.initials: V
authors.lastname: Collazo
authors.forename: C
authors.initials: C
authors.lastname: Alonso
authors.forename: G
authors.initials: G
authors.lastname: Frutos
mesh_terms: D057205:Automation, Laboratory; D002138:Calibration; D002626:Chemistry, Pharmaceutical; D002863:Chromogenic Compounds / Q000737:chemistry; D003029:Coagulants / Q000737:chemistry*; D005169:Factor VIII / Q000032:analysis*; D006801:Humans; D007700:Kinetics; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
chemical_list: D002863:Chromogenic Compounds; D003029:Coagulants; C078147:F8 protein, human; D005169:Factor VIII