Validation of HPLC Method for Determination of Histamine in Human Immunoglobulin Formulations.

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.
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