Makoto Suzuki1, Shin Nishiumi1, Takashi Kobayashi1, Arata Sakai1, Yosuke Iwata2, Takato Uchikata2, Yoshihiro Izumi3,4, Takeshi Azuma1, Takeshi Bamba3,4, Masaru Yoshida1,5,6. 1. Division of Gastroenterology, Department of Internal Medicine, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe, Hyogo, 650-0017, Japan. 2. Shimadzu Corporation, 1 Nishinokyo Kuwabara-cho, Nakagyo-ku, Kyoto, 604-8511, Japan. 3. Medical Institute of Bioregulation, Kyusyu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. 4. Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan. 5. Division of Metabolomics Research, Department of Internal Related, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe, Hyogo, 650-0017, Japan. 6. AMED-CREST, AMED, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe, Hyogo, 650-0017, Japan.
Abstract
RATIONALE: The analytical stability and throughput of biomarker assays based on dried serum spots (DSS) are strongly dependent on the extraction process and determination method. In the present study, an on-line system based on supercritical fluid extraction-supercritical fluid chromatography coupled with tandem mass spectrometry (SFE-SFC/MS/MS) was established for analyzing the levels of disease biomarkers in DSS. METHODS: The chromatographic conditions were investigated using the ODS-EP, diol, and SIL-100A columns. Then, we optimized the SFE-SFC/MS/MS method using the diol column, focusing on candidate biomarkers of oral, colorectal, and pancreatic cancer that were identified using liquid chromatography (LC)/MS/MS. RESULTS: By using this system, four hydrophilic metabolites and 17 hydrophobic metabolites were simultaneously detected within 15 min. In an experiment involving clinical samples, PC 16:0-18:2/16:1-18:1 exhibited 93.8% sensitivity and 64.3% specificity, whereas PC 17:1-18:1/17:0-18:2 showed 81.3% sensitivity and 92.9% specificity for detecting oral cancer. In addition, assessments of the creatine levels demonstrated 92.3% sensitivity and 78.6% specificity for detecting colorectal cancer. CONCLUSIONS: The results of this study indicate that our method has great potential for clinical diagnosis and would be suitable for large-scale screening.
RATIONALE: The analytical stability and throughput of biomarker assays based on dried serum spots (DSS) are strongly dependent on the extraction process and determination method. In the present study, an on-line system based on supercritical fluid extraction-supercritical fluid chromatography coupled with tandem mass spectrometry (SFE-SFC/MS/MS) was established for analyzing the levels of disease biomarkers in DSS. METHODS: The chromatographic conditions were investigated using the ODS-EP, diol, and SIL-100A columns. Then, we optimized the SFE-SFC/MS/MS method using the diol column, focusing on candidate biomarkers of oral, colorectal, and pancreatic cancer that were identified using liquid chromatography (LC)/MS/MS. RESULTS: By using this system, four hydrophilic metabolites and 17 hydrophobic metabolites were simultaneously detected within 15 min. In an experiment involving clinical samples, PC 16:0-18:2/16:1-18:1 exhibited 93.8% sensitivity and 64.3% specificity, whereas PC 17:1-18:1/17:0-18:2 showed 81.3% sensitivity and 92.9% specificity for detecting oral cancer. In addition, assessments of the creatine levels demonstrated 92.3% sensitivity and 78.6% specificity for detecting colorectal cancer. CONCLUSIONS: The results of this study indicate that our method has great potential for clinical diagnosis and would be suitable for large-scale screening.
Authors: Vijayasarathy Ketavarapu; Vishnubhotla Ravikanth; Mitnala Sasikala; G V Rao; Ch Venkataramana Devi; Prabhakar Sripadi; Murali Satyanarayana Bethu; Ramars Amanchy; H V V Murthy; Stephen J Pandol; D Nageshwar Reddy Journal: BMC Cancer Date: 2022-07-19 Impact factor: 4.638
Authors: Francisco W Maciel-Silva; Daniel Lachos-Perez; Luz Selene Buller; William G Sganzerla; Montserrat Pérez; Mauricio A Rostagno; Tania Forster-Carneiro Journal: Molecules Date: 2022-09-23 Impact factor: 4.927