Analytical Sciences

HPLC Determination of Acetaminophen in Saliva Based on Precolumn Fluorescence Derivatization with 12-(3,5-Dichloro-2,4,6-triazinyl)-benzo[d]benzo[1′,2′-6,5]isoindolo[1,2-b][1,3]thiazolidine

Hiroyuki FUJINO,* Hideyuki YOSHIDA,** Hitoshi NOHTA,** and Masatoshi YAMAGUCHI**

*PET Diagnostic Imaging Center, Kitakyusyu Medical Checkup Clinic, 3-1-2 Muromachi, Kokura-Kita, Kitakyusyu 803-0812, Japan
**Faculty of Pharmaceutical Sciences, Fukuoka University, Nanakuma, Johnan, Fukuoka 814-0180, Japan

A high-performance liquid-chromatographic method for the determination of acetaminophen in saliva has been developed. This method is based on the precolumn derivatization of acetaminophen with 12-(3,5-dichloro-2,4,6-triazinyl)benzo[d]benzo[1′,2′-6,5]isoindolo[1,2-b][1,3]thiazolidine, a new fluorescence derivatization reagent for phenolic compounds. The resulting derivative of acetaminophen is separated by isocratic elution on a reversed-phase column, and is fluorometrically detected at an emission wavelength of 560 nm with an excitation wavelength of 540 nm. The detection limit (signal-to-noise ratio = 3) was 0.1 µg/mL in saliva. The proposed method permits a highly sensitive and simple determination of acetaminophen in a small amount of saliva without any sample purification.

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