Method of detecting fluorescent materials and apparatus for their detection

In combination, an analytical apparatus for the quantitative determination of a material selected from the group of polycyclic aromatic hydrocarbons, dansylamino acids and vitamins in a mixture comprising:

• (a) means defining and containing a supply of said materials to be separated;
• (b) a liquid column chromatograph for separating said material to form a column eluate containing the separated said material;
• (c) said material separating means in said chromatographic column;
• (d) a first mixing zone;
• (e) means defining and containing a supply of oxalic acid derivative reagent;
• (f) means for passing a solution of an oxalic acid derivative to said first mixing zone at a controlled rate;
• (g) means defining and containing a supply of hydrogen peroxide reagent;
• (h) means for passing a solution of hydrogen peroxide to said first mixing zone at a controlled rate;
• (i) means for simultaneously introducing the separated solution, oxalic acid derivative reagent and hydrogen peroxide into said first mixing means;
• (j) means in said first mixing zone to mix said column eluate, said oxalic acid derivative, and said hydrogen peroxide to form a first mixture;
• (k) a coiled mixing zone through which said first mixture is continuously passed to form a second mixture, whereby chemiluminescence occurs;
• and
• (l) a light-free detecting section comprising a flow cell and a light receptive detection means for perceiving and multiplying the light produced by the chemiluminescence reaction.