Method of detecting the presence of polycyclic aromatic hydrocarbons
View Patent ↗Described herein is a rapid coupled enzymatic assay to detect polycyclic aromatic hydrocarbon (PAH) compounds in samples. The method uses a detection composition that includes: a buffered solution comprising NADH; naphthalene dioxygenase, naphthalene 1,2-dihydroxy-1,2-dihydronaphthalene dehydrogenase, and 1,2-dihydroxynaphthalene dioxygenase. The presence of PAH is determined by noting a color change of the detection composition, either visually or spectrophotometrically.
1. A method of detecting the presence of polycyclic aromatic hydrocarbons comprising:
adding a sample suspected of containing one or more polycyclic aromatic hydrocarbons to a detection composition, wherein the detection composition comprises: NADH;
naphthalene dioxygenase; naphthalene 1,2-dihydroxy-1,2-dihydronaphthalene dehydrogenase; and 1,2-dihydroxynaphthalene dioxygenase; and
detecting the presence of polycyclic aromatic hydrocarbons by determining if the color of the detection composition has changed due to the presence of polycyclic aromatic hydrocarbons.
2. The method of claim 1 , wherein the sample is derived from food.
3. The method of claim 1 , wherein the sample is obtained from an environment.
4. The method of claim 1 , wherein the naphthalene dioxygenase is stabilized naphthalene dioxygenase.
5. The method of claim 4 , wherein the stabilized naphthalene dioxygenase is produced by the method comprising:
obtaining naphthalene dioxygenase from a bacterial source; and
dialyzing the obtained naphthalene dioxygenase using dialysis tubing having a molecular weight cut off (MWCO) of less than or equal to about 7500.
6. The method of claim 5 , wherein the dialysis is performed in the presence of dithiothreitol (DTT) and/or glycerol.
7. The method of claim 1 , wherein the naphthalene 1,2-dihydroxy-1,2-dihydronaphthalene dehydrogenase is desalted before use in the detection composition.
8. The method of claim 1 , wherein the 1,2-dihydroxynaphthalene dioxygenase is desalted before use in the detection composition.
9. The method of claim 1 , wherein a change in the color of the detection composition is determined by detecting a change in the specific absorbance of ultraviolet or visible light by the detection composition.
10. The method of claim 1 , wherein a change in the color of the detection composition is determined by detecting a change in the specific absorbance at wavelengths of between about 360 nm to about 550 nm.