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2)Laboratory of Proteomics and Biomolecular Science, Biomedical Research Core Facilities, 3)Laboratory of Bioanalytical Chemistry, Tohoku Medical and Pharmaceutical University, Miyagi, Japan1)Department of Esophageal and Gastroenterological Surgery, Juntendo University Graduate School of Medicine, Tokyo, JapanSei MATSUMORI1), Takashi HASHIMOTO1), Motomi NASU1), Naoko KAGA2), Hikari TAKA2), Tsutomu FUJIMURA2, 3), Takashi UENO2), Yoshiki MIURA2), Yoshiaki KAJIYAMA1)Corresponding author: Takashi Hashimoto Department of Esophageal and Gastroenterological Surgery, Juntendo University Graduate School of Medicine2-1-1 Hongo Bunkyo-ku, Tokyo 113-8421, JapanTEL: +81-3-3813-3111　FAX: +81-3-5802-1951　E-mail: tks-hsmt@juntendo.ac.jp〔Received　May. 21, 2022〕〔Accepted　Jul. 8, 2022〕J-STAGE Advance published date: Oct. 4, 2022Copyright © 2022 The Juntendo Medical Society. This is an open access article distributed under the terms of Creative Commons Attribution License (CC BY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original source is properly credited. doi: 10.14789/jmj.JMJ22-0023-OAObjectives: Since esophageal carcinoma progresses asymptomatically, for many patients the disease is already advanced at the time of diagnosis. The main methods that are currently used to diagnose esophageal carcinoma are upper gastrointestinal radiographic contrast examinations and upper gastrointestinal endoscopy, but early discovery of this disease remains difficult. There is a need to develop a diagnostic method using biomarkers that is non-invasive while both highly sensitive and specific.Materials and Methods: Exhaled breath was collected from 17 patients with esophageal squamous cell carcinoma (ESCC), as well as 9 control subjects without history of any cancer. For each fasting subject, 1L of exhaled breath was collected in a gas sampling bag. Volatile organic compounds (VOCs) were then extracted from each sample using Solid phase micro-extraction (SPME) fibers and analyzed by gas chromatography-mass spectrometry (GC-MS).Results: Levels of acetonitrile, acetic acid, acetone, and 2-butanone in exhaled breath were significantly higher in the patient group than in the control group (p = 0.0037, 0.0024, 0.0024 and 0.0037, respectively). ROC curves were drawn for these 4 VOCs, and the results for the area-under-the-curve (AUC) indicated that ESCC patients can be identified with a high probability of 0.93.Conclusion: We found distinctive VOCs in exhaled breath of ESCC patients. These VOCs have a potential as new clinical biomarkers for ESCC. The measurement of VOCs in exhaled breath may be a useful, non-invasive method for diagnosis of ESCC.Key words: Volatile organic compounds (VOCs), Esophageal squamous cell carcinoma (ESCC), Gas chromatography-mass spectrometry (GC-MS)Juntendo Medical Journal2022. 68(5), 499-504Original ArticlesDevelopment of a Non-invasive Diagnostic Method for Esophageal Squamous Cell Carcinoma IntroductionEsophageal carcinoma is the tenth most common malignancy in the world1), and the tenth most common cause of cancer death in Japan2）. Incidence of squamous cell carcinoma of the esophagus (ESCC) is decreasing in Europe and America, whereas the incidence of esophageal adenocarci-noma is, conversely, trending upward. In Asia, squamous cell carcinoma still accounts for a majority Juntendo University Graduate School of Medicine, Tokyo, Japanof all esophageal carcinomas3). Known risk factors for ESCC include smoking, drinking alcoholic beverages, hot foods, drinks and malnutrition4). The quality of diagnosis and treatment has improved recently, but these cancers still have a poor prog-nosis. The main symptoms of esophageal carcinoma are dysphagia and a feeling that food is stuck in the throat. However, because only advanced cancers cause these symptoms, there are usually no symp-toms in the early stage of the disease. As a result, 499by Gas Chromatographic Analysis of Exhaled Breath