title: Estimation of uncertainty from method validation data: Application to a reverse-phase high-performance liquid chromatography method for the determination of amino acids in gelatin using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent.
abstract: A detailed procedure for estimating uncertainty according to the Laboratory of Government Chemists/Valid Analytical Measurement (LGC/VAM) protocol for determination of 18 amino acids in gelatin is proposed. The expanded uncertainty was estimated using mainly the method validation data (precision and trueness). Other sources of uncertainties were contributed by components in standard preparation measurements. The method scope covered a single matrix (gelatin) under a wide range of analyte concentrations. The uncertainty of method precision, μ(P) was 0.0237-0.1128pmolμl(-1) in which hydroxyproline and histidine represented the lowest and highest values of uncertainties, respectively. Proline and phenylalanine represented the lowest and highest uncertainties value for method recovery, μ(R) that was estimated within 0.0064-0.0995pmolμl(-1). The uncertainties from other sources, μ(Std) were 0.0325, 0.0428 and 0.0413pmolμl(-1) that were contributed by hydroxyproline, other amino acids and cystine, respectively. Hydroxyproline and phenylalanine represented the lowest and highest values of expanded uncertainty, U(y) that were determined at 0.0949 and 0.2473pmolμl(-1), respectively. The data were accurately defined and fulfill the technical requirements of ISO 17025:2005.
authors.affiliation: Laboratory of Halal Analysis, Halal Products Research Institute, Universiti Putra Malaysia, 43400 Selangor, Malaysia.
authors.forename: M I
authors.initials: MI
authors.lastname: Azilawati
authors.affiliation: Laboratory of Halal Analysis, Halal Products Research Institute, Universiti Putra Malaysia, 43400 Selangor, Malaysia.
authors.forename: M H
authors.initials: MH
authors.lastname: Dzulkifly
authors.affiliation: Laboratory of Halal Analysis, Halal Products Research Institute, Universiti Putra Malaysia, 43400 Selangor, Malaysia.
authors.forename: B
authors.initials: B
authors.lastname: Jamilah
authors.affiliation: Laboratory of Halal Analysis, Halal Products Research Institute, Universiti Putra Malaysia, 43400 Selangor, Malaysia.
authors.forename: M
authors.initials: M
authors.lastname: Shuhaimi
authors.affiliation: Laboratory of Halal Analysis, Halal Products Research Institute, Universiti Putra Malaysia, 43400 Selangor, Malaysia; Department of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Selangor, Malaysia. Electronic address: aminis@upm.edu.my.
authors.forename: I
authors.initials: I
authors.lastname: Amin
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D000634:Aminoquinolines / Q000737:chemistry*; D002219:Carbamates / Q000737:chemistry*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D005780:Gelatin / Q000737:chemistry*; D006909:Hydroxyproline / Q000737:chemistry; D007202:Indicators and Reagents / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results; D035501:Uncertainty
chemical_list: C078178:6-aminoquinolyl-N-hydroxysuccinimidyl carbamate; D000596:Amino Acids; D000634:Aminoquinolines; D002219:Carbamates; D007202:Indicators and Reagents; D005780:Gelatin; D006909:Hydroxyproline
keywords: 6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent; Amino acids; Estimation of uncertainty; Gelatin