Development of LC-MS/MS-based accurate quantification methods for assessing serum vitamin D metabolites

• Project/Area Number: 15H06096
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Laboratory medicine
• Research Institution: Chiba University
• Principal Investigator: ISHIGE Takayuki 千葉大学, 医学部附属病院, 臨床検査技師 (30757315)
• Research Collaborator: SATOH Mamoru 千葉大学, 医学部附属病院, 特任准教授 (20401002) ; NOMURA Fumio 千葉大学, 医学部附属病院, 特任教授 (80164739) ; NISHIMURA Motoi 千葉大学, 医学部附属病院, 講師 (80400969) ; OGAWA Shoujiro 東京理科大学, 薬学部, 助教 (30546271) ; HIGASHI Tatsuya 東京理科大学, 薬学部, 教授 (90272963)
• Project Period (FY): 2015-08-28 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: ビタミンD代謝物 / LC-MS/MS / 誘導体化

Outline of Final Research Achievements

DAPTAD derivatization of vitamin D metabolites were performed for enhancing the ionization efficiency of LC-MS/MS. As compared with 25(OH)D3, the intensity of 25(OH)D3-DAPTAD was increased approximately 100-fold higher. By using SLE for sample preparation, the high-sensitive and robust method for quantification of 4 serum vitamin D metabolites was established. We are planning to establish serum 1,25(OH)2D analysis method by LC/MS/MS.

Research Products

• [Journal Article] Locked nucleic acid probe enhances Sanger sequencing sensitivity and improves diagnostic accuracy of high-resolution melting-based KRAS mutational analysis (2016)
  • Author(s): Takayuki Ishige, Sakae Itoga, Kazuyuki Matsushita, Fumio Nomura
  • Journal Title: Clinica Chimica Acta Volume: 457 Pages: 75-80
  • DOI: 10.1016/j.cca.2016.04.005
  • Peer Reviewed
• [Journal Article] Development and validation of the simultaneous measurement of four vitamin D metabolites in serum by LC-MS/MS for clinical laboratory applications (2016)
  • Author(s): Mamoru Satoh, Takayuki Ishige, Shoujiro Ogawa, Motoi Nishimura, Kazuyuki Matsushita, Tatsuya Higashi, Fumio Nomura
  • Journal Title: Anal Bioanal Chem Volume: 408 Issue: 27 Pages: 7617-7627
  • DOI: 10.1007/s00216-016-9821-4
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Combined Secretomics and Transcriptomics Revealed Cancer-Derived GDF15 is Involved in Diffuse-Type Gastric Cancer Progression and Fibroblast Activation. (2016)
  • Author(s): Ishige T, Nishimura M, Satoh M, Fujimoto M, Fukuyo M, Semba T, Kado S, Tsuchida S, Sawai S, Matsushita K, Togawa A, Matsubara H, Kaneda A, Nomura F
  • Journal Title: Sci Rep. Volume: 6 Issue: 1 Pages: 21681-21681
  • DOI: 10.1038/srep21681
  • Peer Reviewed / Open Access
• [Journal Article] Assessment of Vitamin D Metabolism by LC-MS/MS (2015)
  • Author(s): Ishige T, Satoh M, Ogawa S, Nishimura M, Higashi T, Nomura F.
  • Journal Title: Rinsho Byori Volume: 63 Pages: 457-464
  • NAID: 40020474101
• [Presentation] サンガー法の検出感度向上のための野生型アレル阻害サイクルシーケンス反応 (2016)
  • Author(s): 石毛崇之、糸賀栄、西村基、野村文夫、松下一之
  • Organizer: 第23回日本遺伝子診療学会
  • Place of Presentation: イイノホール＆カンファレンスセンター（東京都・千代田区）
  • Year and Date: 2016-10-08
• [Presentation] Long-PCRと大量並列シーケンシングによる家族性地中海熱の原因遺伝子MEFVの遺伝学的検査 (2016)
  • Author(s): 石毛崇之、糸賀栄、北村浩一、西村基、松下一之、野村文夫
  • Organizer: 日本臨床検査自動化学会第48回大会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-09-22
• [Presentation] Integrated secretome and transcriptome analyses for identification of functional secreted molecules involved in diffuse-type gastric cancer progression (2016)
  • Author(s): Takayuki Ishige, Motoi Nishimura, Mamoru Satoh, Kazuyuki Matsushita, Fumio Nomura
  • Organizer: The association for Mass Spectrometry: Applications to the Clinical Lab 2016 EU (MSACL2016EU)
  • Place of Presentation: Salzburg Congress Center, AUSTRIA
  • Year and Date: 2016-09-15
• [Presentation] LC/MS/MSによる血清ビタミンD代謝物の精密分析 (2016)
  • Author(s): 石毛崇之
  • Organizer: 千葉県臨床検査技師会第25回千葉地区研修会
  • Place of Presentation: 千葉メディカルセンター（千葉県・千葉市）
  • Year and Date: 2016-07-02
• [Presentation] LC/MS/MSの臨床検査応用 - ビタミンD代謝物測定を基盤に - (2015)
  • Author(s): 佐藤守, 石毛崇之, 小川祥二郎, 西村基, 松下一之, 東達也, 野村文夫
  • Organizer: 日本臨床検査自動化学会第47回大会
  • Place of Presentation: パシフィコ横浜 (神奈川県･横浜市)
  • Year and Date: 2015-10-08
• [Presentation] LNA-PCR Sequencingによる高感度KRAS遺伝子変異解析 (2015)
  • Author(s): 石毛崇之, 糸賀栄, 松下一之, 野村文夫
  • Organizer: 日本臨床検査自動化学会第47回大会
  • Place of Presentation: パシフィコ横浜 (神奈川県･横浜市)
  • Year and Date: 2015-10-08
• [Presentation] LC/MS/MSの臨床検査応用 - ビタミンD代謝物測定を基盤に - (2015)
  • Author(s): 佐藤守, 石毛崇之, 小川祥二郎, 西村基, 松下一之, 東達也, 野村文夫
  • Organizer: 第40回日本医用マススペクトル学会年会
  • Place of Presentation: アクトシティ浜松コングレスセンター (静岡県･浜松市)
  • Year and Date: 2015-09-17