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A liquid chromatographic method for the determination of histamine in immunoglobulin preparation using solid phase extraction and pre-column derivatization

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An immunoglobulin (IgG) preparation with micro-amount of histamine fixed on the active protein fraction has been used to increase the resistance to allergic reactions. However, excessive histamine may cause hypo- or hypertension, headache, or anaphylactic shock and so the histamine content of the drug is strictly controlled by a regulation: 0.15 μg of histamine dihydrochloride is allowed for 12 mg of immunoglobulin. In this study, a liquid Chromatographic method to determine micro-amount of histamine in the pharmaceutical was developed and validated. This method include a sample cleanup by a solid phase extraction (SPE) using a polystyrenedivinyl benzene (PS-DVB) polymeric sorbent and high-performance liquid chromatography after precolumn fluorescent labeling of the histamine with o-phthaldialdehyde. The drug sample was loaded to the SPE cartridge after adjusting to pH 9.5. After successive washings of the cartridge with water and 30% aqueous methanol, histamine was then eluted with 100 mM sodium acetate (pH 9.5)-methanol (20:80, v/v). An aliquot from the eluate was labeled with o-phthaldialdehyde-mercaptoethanol (OPA-ME) for fluorescence detection at the excitation maximum of 340 nm and emission maximum of 450 nm. HPLC analysis was performed on a phenyl-hexyl column with an acetonitrile-phosphate buffer (pH 6.8; 50 μM) (35:65, v/v) as the mobile phase. The retention times of histamine and 3-methylhistamine (IS) were approximately 7.2 and 9.5 min, respectively. The quantitation range was between 0.01-0.2 mg/mL of histamine showing good linearity (r=0.9996). This analytical method would provide a potential mean for the strict control of histamine content in the pharmaceutical product.

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Correspondence to Nam Hee Kim or Youmie Park or Eun Sook Jeong or Chang-Soo Kim or Min Kyo Jeoung or Kyoung Soon Kim or Seung-Hwa Hong or Jong-Keun Son or Jin Tae Hong or II-young Park or Dong-Cheul Moon.

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Kim, N.H., Park, Y., Jeong, E.S. et al. A liquid chromatographic method for the determination of histamine in immunoglobulin preparation using solid phase extraction and pre-column derivatization. Arch. Pharm. Res. 30, 1350 (2007). https://doi.org/10.1007/BF02980277

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Key words

  • Immunoglobulin preparation
  • Histamine
  • Solid-phase extraction
  • PS-DVB
  • Fluorescent detection
  • o-Phthaldialdehyde