Determination of Formaldehyde in the Polymerized Ragweed Antigen by HPLC

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Abstract
Formaldehyde is used in the polymerization of ragweed antigen for the desensitization treatment of hay fever. Because the polymerized ragweed antigen is used as a vaccine which stimulates the production of the desired immunological effects upon injecting into the body, trace formaldehyde in the injected material is a health concern. Two procedures for the determination of formaldehyde were developed. In the direct procedure, formaldehyde in the sample was reacted with 2,4-dinitrophenylhydrazine (2,4-DNP) in solution to yield a strongly UV absorbing derivative. In an alternate approach, formaledhyde in the sample in a sealed vial was allowed to diffuse in the gaseous form into a reaction chamber consisted of a culture tube insert in which formaldehyde was derivatized with 2,4-DNP reagent. Following extraction into methylene chloride, the derivative was injected into the liquid chromatograph, separated on a reversed phase column and detected at UV 254 nm.