ABSTRACT
BACKGROUND: Histamine fixed-immunoglobulin formulations, which consisted of 0.15 µg of histamine dihydrochloride and 12 mg of human immunoglobulin in a vial, are used for anti-allergic treatments, and controlling the amounts of histamine in the formulations is essential to avoid histamine intoxication. OBJECTIVE: A high-performance liquid chromatography (HPLC) method for determination of histamine contents of the formulations was established and validated. METHODS: Histamine extracted from the formulation was labeled with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate and was analyzed by gradient elution HPLC with UV detection at 260 nm. RESULTS: The method showed linearity in the range 0.8-2.4 µM (R > 0.999), accuracy (100.1-105.8% recovery), and precision (relative standard deviation ≤ 1.93%). The validated method was applied for five lots of the pharmaceutical, and their histamine contents were determined to be 0.149-0.155 µg/vial. CONCLUSIONS: These results indicated that the validated method is useful to control amounts of histamine in biopharmaceutical products. HIGHLIGHTS: The HPLC method was developed for quantitative determination of histamine content of the histamine fixed-immunoglobulin formulations.
