Validation of the quantification of histamine in rat brain using the radioenzymatic assay

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Quantification of histamine (Hm) in rat brain is problematic. The authors have evaluated tissue extraction procedures and validated a new Hm radioenzymatic assay for brain. Brain was extracted twice with either 0.1M perchloric acid (PCA), boiling water, or water. Sacrifice by microwave irradiation increased whole brain Hm from 64.4 +/- 16.5 to 450.9 +/- 90.8 ng/g. The validity of the assay was demonstrated by isolating the product formed by the radioenzymatic assay and subjecting it to TLC and autoradiography. The sensitivity of this assay permitted the quantification of Hm in as little as 4.3 ..mu..g of hypothalamus. The authors conclude: 1) The preferred method for sacrifice is decapitation; 2) homogenization in PCA effectively extracts Hm from brain tissue; 3) the radioenzymatic assay is specific for Hm in this tissue.

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