title: Improved HPLC Method Using 2,3-naphthalenedicarboxaldehyde as Fluorescent Labeling Agent for Quantification of Histamine in Human Immunoglobulin Preparations.
abstract: OBJECTIVE
  To develop and optimize quantitative HPLC method using 2,3-naphthalenedicarboxaldehyde (NDA) after simple and efficient solid phase extraction to determine the histamine in a biopharmaceutical (Histobulin™).
  
  
  METHODS
  The HPLC method was established using NDA-induced Histobulin and compared with the recently reported HPLC method using o-phthaldehyde (OPA). The validated NDA-applied HPLC method was adjusted to 15 lots of Histobulin and compared by the current lot-release-test method using fluorimetry in recovery of histamine and reproducibility.
  
  
  RESULTS
  Analyses of six HPLC chromatograms using NDA and OPA each were compared. NDA produced a more stable chromatogram baseline than OPA, and showed better stability. The HPLC analysis was validated in accuracy (91-103%), precision (interday/intraday assay CV ≤2.30%), and linearity of dose-response curve (R(2) ≥ 0.9919). The detection limit was 0.0076 μg/mL and the quantitative limit was 0.0229 μg/mL. The amount of histamine per 12 mg of immunoglobulin was determined to be 0.17 ± 0.016 μg by the HPLC and 0.025 ± 0.013 μg by the current lot-release-test method using fluorimetry.
  
  
  CONCLUSIONS
  NDA derivatization showed better stability compared with the OPA method. Therefore the newly established NDA-derivatizated HPLC method may be more suitable than the fluorimetric method in lot-release-tests of biopharmaceuticals.
authors.affiliation: Blood Products Teams, National Institute of Food and Drug Safety Evaluation, Osong, Korea.
authors.forename: Jung-Hwan
authors.initials: JH
authors.lastname: Kim
authors.forename: In Soo
authors.initials: IS
authors.lastname: Shin
authors.forename: Yoo Kyoung
authors.initials: YK
authors.lastname: Lee
authors.forename: Ho Jung
authors.initials: HJ
authors.lastname: Oh
authors.forename: Sang Ja
authors.initials: SJ
authors.lastname: Ban
keywords: 2,3-naphthalenedi-carboxaldehye (NDA); high-performance liquid chromatography (HPLC); histamine; immunoglobulin (Ig) preparation; lot-release test