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Biogenic amine assay using HPLC-ECD

  • US 5,011,608 A
  • Filed: 11/18/1988
  • Issued: 04/30/1991
  • Est. Priority Date: 11/18/1988
  • Status: Expired due to Fees
First Claim
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1. A method for the assaying of low concentrations of an oxidizable or reducible biogenic amine or derivative or metabolite thereof, comprising the steps of:

  • (a) preparing a sample of at least one said compound, said sample being suitable for quantitative analysis;

    (b) injecting said sample into a high pressure liquid chromatography apparatus to thereby form a chromatogram, said apparatus including;

    (1) a chromatography column containing a quantity of microparticulate material as a stationary phase;

    (2) a mobile phase passing through said stationary phase, said mobile phase comprising the following components;

    (a) from about 50 mM to about 250 mM of a buffer having a pH of from about 2 to about 8;

    (b) from about 0.1 mM to about 10.0 mM of a chelating agent;

    (c) from about 0.1 mM to about 10.0 mM of an alkyl sulfonate, said alkyl sulfonate having from about 4 to about 14 carbon atoms; and

    (d) from about 0.1% to about 10% by volume methanol in water solution;

    (3) an injector for injecting said sample;

    (4) a pump for pumping said mobile phase through said stationary phase; and

    (5) a guard column positioned upstream of said chromatography column;

    (c) eluting said compounds from said chromatography column;

    (d) passing said eluted compounds through an electrochemical detector cell thereby producing a current output from said detector;

    (e) measuring and integrating the current output from said electrochemical detector to obtain a quantitative analysis of said compound; and

    (f) maintaining a substantially constant base line noise at a sensitivity of said electrochemical detector cell sufficient to measure the quantity of said compound present in resting, human plasma by maintaining all portions of said apparatus contacting said mobile phase and said stationary phase substantially residue-free, wherein the signal to noise ratio of said current output is at least 2 to 1.

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