title: Determination of acetone in saliva by reversed-phase liquid chromatography with fluorescence detection and the monitoring of diabetes mellitus patients with ketoacidosis.
abstract: BACKGROUND
  In diabetes mellitus (DM) patients with ketoacidosis, ketone bodies, i.e., acetone, acetoacetic acid (AA) and β-hydroxybutyric acid (HA), are increased in the blood and urine. Acetone is also excreted by breathing due to the spontaneous decomposition of AA. Thus, the increase in acetone has been considered as one of the biomarkers for the diagnosis of DM. However, the determination of acetone in one's breath is not recommended because of the sample handling difficulty. We measured acetone in saliva by reversed-phase liquid chromatography (LC) with fluorescence (FL) detection. The proposed method was applied to the determination of acetone in the saliva of healthy volunteers and DM patients with and without ketoacidosis.
  
  
  METHODS
  3-Pentanone (I.S.) and DBD-H in acetonitrile were added to freshly collected saliva and reacted at room temperature for 20 min in the presence of trifluoroacetic acid. After the reaction, the solution was centrifuged at 10,000 × g and 4 °C for 5 min. The supernatant was separated by reversed-phase LC and the FL detected at 550 nm (excitation at 460 nm).
  
  
  RESULTS
  The concentrations of acetone in the DM patients with ketoacidosis were significantly higher than those of the normal subjects and DM patients without ketoacidosis. Furthermore, the total contents of the ketone bodies in the blood correlated with acetone in the saliva of the DM patients. The concentrations of acetone in the saliva of an emergency patient also correlated with the ketone bodies in the blood at each sampling time.
  
  
  CONCLUSIONS
  The proposed method using LC-FL seems to be useful for the determination of acetone in the saliva of DM patients with ketoacidosis. The method offers a new option for the diagnosis and monitoring of DM patients with ketoacidosis.
authors: Laboratory of Analytical and Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors: Shinya
authors: S
authors: Fujii
authors: Laboratory of Clinical Pharmaceutics and Pharmacy Practice, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors: Toshio
authors: T
authors: Maeda
authors: Department of Pharmaceutical, Numazu City Hospital, Harunoki Higashi-Shiiji Numazu 410-0302, Japan.
authors: Ichiro
authors: I
authors: Noge
authors: Department of Clinical and Molecular Endocrinology, Numazu City Hospital, Harunoki Higashi-Shiiji Numazu 401-0302, Japan.
authors: Yutaka
authors: Y
authors: Kitagawa
authors: Laboratory of Analytical and Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors: Kenichiro
authors: K
authors: Todoroki
authors: Laboratory of Analytical and Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors: Koichi
authors: K
authors: Inoue
authors: Laboratory of Analytical and Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors: Jun Zhe
authors: JZ
authors: Min
authors: Laboratory of Analytical and Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan. Electronic address: toyooka@u-shizuoka-ken.ac.jp.
authors: Toshimasa
authors: T
authors: Toyo'oka
mesh_terms: D000096:Acetone / Q000032:analysis*; D000328:Adult; D000368:Aged; D056148:Chromatography, Reverse-Phase; D003920:Diabetes Mellitus / Q000175:diagnosis / Q000378:metabolism*; D005260:Female; D005453:Fluorescence; D006801:Humans; D007662:Ketosis / Q000150:complications* / Q000175:diagnosis / Q000378:metabolism*; D008163:Luminescent Measurements; D008297:Male; D008875:Middle Aged; D012463:Saliva / Q000737:chemistry*
chemical_list: D000096:Acetone
keywords: Acetone; Diabetes mellitus; Fluorescence labeling; Ketoacidosis; Liquid chromatography; Saliva