Application of 13C stable isotope labeling liquid chromatography-multiple reaction monitoring-tandem mass spectrometry method for determining intact absorption of bioactive dipeptides in rats

Eduardo M.N. Nakashima, Akihiro Kudo, Yuri Iwaihara, Mitsuru Tanaka, Kiyoshi Matsumoto, Toshiro Matsui

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The liquid chromatography-multiple reaction monitoring-tandem mass spectrometry (LC-MRM-MS/MS) method using 13C stable isotope-labeled dipeptides was newly developed to simultaneously determine the absorption of three antihypertensive peptides (Val-Tyr, Met-Tyr, and Leu-Tyr) into blood of spontaneously hypertensive rats in one run-in assay. After extracting 13C-labeled peptides in blood sample with a C18 cartridge, the extract was applied to a 13C monoisotopic transition LC-MRM-MS/MS system with d-Val-Tyr included as internal standard. An excellent separation of each dipeptide in LC was achieved at the elution condition of 5-100% methanol in 0.1% formic acid at a flow rate of 0.25 ml/min. The 13C-labeled peptides ionized by electron spray were detected in the positive ion mode within 15 min. The established method showed high reproducibility with less than 10% coefficient of variation as well as high accuracy of more than 85%. After the administration of a mixture containing the three 13C-labeled dipeptides to rats at each dose of 30 mg/kg, we could successfully determine the intact absorption of each 13C- labeled peptide with the maximal absorption amount of 1.1 ng/ml plasma for Val-Tyr by the proposed LC-MRM-MS/MS method.

Original languageEnglish
Pages (from-to)109-116
Number of pages8
JournalAnalytical Biochemistry
Volume414
Issue number1
DOIs
Publication statusPublished - Jul 1 2011

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valyltyrosine
Isotope Labeling
Dipeptides
Liquid chromatography
Tandem Mass Spectrometry
Liquid Chromatography
Isotopes
Labeling
Mass spectrometry
Rats
Peptides
formic acid
Monitoring
leucyltyrosine
Blood
Inbred SHR Rats
Antihypertensive Agents
Methanol
Assays
Positive ions

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Application of 13C stable isotope labeling liquid chromatography-multiple reaction monitoring-tandem mass spectrometry method for determining intact absorption of bioactive dipeptides in rats. / Nakashima, Eduardo M.N.; Kudo, Akihiro; Iwaihara, Yuri; Tanaka, Mitsuru; Matsumoto, Kiyoshi; Matsui, Toshiro.

In: Analytical Biochemistry, Vol. 414, No. 1, 01.07.2011, p. 109-116.

Research output: Contribution to journalArticle

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