Development of a reliable analytical method for endogenous hormones applicable to various animal species

• Project/Area Number: 26550048
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Environmental impact assessment
• Research Institution: Ehime University
• Principal Investigator: Kunisue Tatsuya 愛媛大学, 沿岸環境科学研究センター, 教授 (90380287)
• Co-Investigator(Renkei-kenkyūsha): TANABE SHINSUKE 愛媛大学, 沿岸環境科学研究センター, 教授 (60116952)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2016: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 甲状腺ホルモン / 機器分析 / 遊離型 / タンパク結合型 / ステロイドホルモン

Outline of Final Research Achievements

The present study aimed to develop a sensitive and reliable analytical method for endogenous hormones, especially free thyroid hormones (THs) that play important role in the brain development, in serum/plasma of various animal species using ultrafiltration (UF) in combination with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), which can resolve the interferences encountered due to the nonspecific reactivity of anti-hormone antibodies used in immunoassay (IA) methods. UF at 1100 × g and 37°C for 30 minutes with a 30 kDa ultrafiltration device (Centrifree YM-30) yielded excellent precision. Acceptable accuracy (recovery: 100-110%) and intra- and inter-day precision (CV: <10% and <12%, respectively) were obtained, when triplicate analyses in three different days were conducted using bovine serum. The developed analytical method was successfully applied to the determination of free THs in plasma samples of humans and cats.

Research Products

• [Journal Article] Organohalogens and their hydroxylated metabolites in the blood of pigs from an open waste dumping site in south India: Association with hepatic cytochrome P450 (2015)
  • Author(s): Mizukawa H, Nomiyama K, Kunisue T, Watanabe MX, Subramanian A, Iwata H, Ishizuka M, Tanabe S
  • Journal Title: Environ Res Volume: 138 Pages: 255-263
  • DOI: 10.1016/j.envres.2015.02.012
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Occurrence of Perchlorate and Thiocyanate in Human Serum From E-Waste Recycling and Reference Sites in Vietnam : Association With Thyroid Hormone and Iodide Levels (2014)
  • Author(s): Akifumi Eguchi, Tatsuya Kunisue, Qian Wu, Pham Thi Kim Trang, Pham Hung Viet, Kurunthachalam Kannan, Shinsuke Tanabe
  • Journal Title: Archives of environmental contamination and toxicology Volume: (in press) Issue: 1 Pages: 29-41
  • DOI: 10.1007/s00244-014-0021-y
  • Peer Reviewed
• [Presentation] A sensitive and reliable analytical method for free thyroid hormones in animal serum using ultrafiltration in combination with UHPLC-MS/MS (2016)
  • Author(s): Tanoue, R., Kume, I., Yamamoto, Y., Nomiyama, K., Tanabe, S., Kunisue, T.
  • Organizer: SETAC North America 37th Annual Meeting/7th SETAC World Congress
  • Place of Presentation: Orlando, USA
  • Year and Date: 2016-11-06
  • Int'l Joint Research
• [Presentation] Accumulation features of organohalogen compounds and their hydroxylated metabolites in pet cats and dogs: effects on thyroid hormones homeostasis (2016)
  • Author(s): Nomiyama, K., Yamamoto, Y., Mizukawa, H., Eguchi, A., Nakayama, S. M., Ikenaka, Y., Ishizuka, M., Kunisue, T., Tanabe, S.
  • Organizer: SETAC North America 37th Annual Meeting/7th SETAC World Congress
  • Place of Presentation: Orlando, USA
  • Year and Date: 2016-11-06
  • Int'l Joint Research
• [Presentation] Contamination status of organohalogen compounds and their hydroxylated metabolites in pet cats and dogs: effects on thyroid hormones homeostasis (2016)
  • Author(s): Nomiyama, K., Yamamoto, Y., Mizukawa, H., Takaguchi, K., Takiguchi, M., Eguchi, A., Nakayama, S., Ikenaka, Y., Ishizuka, M., Kunisue, T., Tanabe, S.
  • Organizer: 36th International Symposium on Halogenated Persistent Organic Pollutants
  • Place of Presentation: Firenze, Italy
  • Year and Date: 2016-08-28
  • Int'l Joint Research
• [Presentation] 有機ハロゲン化合物によるコンパニオンアニマルの汚染実態と甲状腺ホルモンへの影響評価 (2016)
  • Author(s): 野見山 桂・山本恭央・水川葉月・江口哲史・滝口満喜・中山翔太・池中良徳・石塚 真由美・国末 達也・田辺信介
  • Organizer: 第25回環境化学討論会
  • Place of Presentation: 新潟コンベンションセンター（新潟県・新潟市）
  • Year and Date: 2016-06-08
• [Presentation] Contamination status of organohalogen compounds and their hydroxylated metabolites in pet cats: Effects on thyroid hormones homeostasis (2015)
  • Author(s): Yamamoto, Y., Nomiyama, K., Mizukawa, H., Takiguchi, M., Ishizuka, M., Kunisue, T. Tanabe, S.
  • Organizer: Society of Environmental Toxicology and Chemistry (SETAC) North America 36th Annual Meeting
  • Place of Presentation: Salt Lake City, Utah, USA
  • Year and Date: 2015-11-01
  • Int'l Joint Research
• [Presentation] 遊離型甲状腺ホルモンの高精度機器分析法の開発 (2015)
  • Author(s): 久米伊万里・田上瑠美・田辺信介・国末達也
  • Organizer: 第24回環境化学討論会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2015-06-24
• [Presentation] ペットネコにおける有機ハロゲン化合物および水酸化代謝物の汚染実態と甲状腺ホルモン恒常性への影響 (2015)
  • Author(s): 山本恭央・野見山 桂・水川葉月・滝口満喜・石塚真由美・国末達也・田辺信介
  • Organizer: 第24回環境化学討論会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2015-06-24