腦脊髓液内의 Norepinephrine，Epinephrine 및 Dopamine 의 定量法

Direct Rapid Radioenzymatic Assay for Norepinephrine, Epinephrine， and Dopamine Simultaneously by a Combination of High Pressure Liquid Chromatography in Human Cerebrospinal Fluid

A direct rapid radioenzymatic assay for measuring simultaneously picogram quantities (2—5 pg) of norepinephrine, epinephrine and dopamine by a high pressure liquid chromatography has been developed. Fifty micrcliters of CSF was incubated with catechol-O-methyltransferase and S-adencsyl-L- 〔methyl-sH〕methionine to convert the three catecholamines to their O-methylated analogues. These derivatives were purified by the solvent extraction and isolated by high pressure liquid chromatography using ᄍ-Bondapak-Cis column and 0.17 M acetic acid-5% methanol—0.02 M Na2 SO4 as eluant. When the integration mark was automatically placed on the chart of data module, manual collection was started two seconds later, and each peak was collected. The norepinephrine and epinephrine derivatives fractions were oxidized to vanillin, and then the solvent extraction of the vanillin and the dopamine derivative fractions were performed and counted by liquid scintillation. A rapid high pressure liquid chromatographic separation of the three catecholamine derivatives, together with the above oxidation and the solvent extraction contributes to high sensitivity and specificity of the method and makes us to complete the differential analysis of 10 CSF samples (20 tubes) within one day. In case of norepinephrine assay alone, more than 30 samples can be completed in a day.