The Chemical Reaction
The "central" chemical in this reaction is luminol (C8H7O3N3), a powdery compound made up of nitrogen, hydrogen, oxygen and carbon. Criminalists mix the luminol powder with a liquid containing hydrogen peroxide (H2O2), a hydroxide (OH-) and other chemicals, and pour the liquid into a spray bottle. The hydrogen peroxide and the luminol are actually the principal players in the chemical reaction, but in order to produce a strong glow, they need a catalyst to accelerate the process. The mixture is actually detecting the presence of such a catalyst, in this case the iron in hemoglobin (see Microsoft Encarta: Catalysis for more information on catalysts).
To perform a luminol test, the criminalists simply spray the mixture wherever they think blood might be. If hemoglobin and the luminol mixture come in contact, the iron in the hemoglobin accelerates a reaction between the hydrogen peroxide and the luminol. In this oxidation reaction, the luminol loses nitrogen and hydrogen atoms and gains oxygen atoms, resulting in a compound called 3-aminophthalate. The reaction leaves the 3-aminophthalate in an energized state -- the electrons in the oxygen atoms are boosted to higher orbitals. The electrons quickly fall back to a lower energy level, emitting the extra energy as a light photon (see How Fluorescent Lamps Work for more information on light production). With iron accelerating the process, the light is bright enough to see in a dark room.
Investigators may use other chemiluminescent chemicals, such as fluorescein, instead of luminol. These chemicals work the same basic way, but the procedure is a little bit different.