Determination of pineal melatonin by precolumn derivatization reversed- phase high-performance liquid chromatography and its application to the study of circadian rhythm in rats and mice

Kenji Hamase, Tatsunosuke Tomita, Ayako Kiyomizu, Kiyoshi Zaitsu

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Determination of minute amounts of endogenous melatonin in rat and mouse pineal gland was performed using an RP-HPLC system. Melatonin was separated following precolumn derivatization and determined with a fluorescence detector at the emission wavelength of 380 nm with the excitation at 245 nm. The calibration curve of melatonin constructed by adding known amounts of melatonin to the homogenates of mouse pineal gland was linear over the range of 1-500 fmol (injection amount/20 μl). The detection limit of added melatonin was 1 fmol (S/N = 5). Repeatability and day-to-day precision for the melatonin spiked sample of mouse pineal gland was 4.0 and 3.8% (RSD), respectively. Using the present method, circadian changes of melatonin content in rat (Wistar) and mouse (C3H) pineal gland were determined. In addition, a minute amount of melatonin in ddY mouse pineal gland was determined, because pineal melatonin of many inbred mouse strains has been reported to be lower than the detection limit. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)106-110
Number of pages5
JournalAnalytical Biochemistry
Volume279
Issue number1
DOIs
Publication statusPublished - Mar 1 2000

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High performance liquid chromatography
Reverse-Phase Chromatography
Melatonin
Circadian Rhythm
Rats
High Pressure Liquid Chromatography
Pineal Gland
Limit of Detection
Inbred Strains Mice
Inbred C3H Mouse
Calibration
Wistar Rats
Fluorescence
Detectors
Wavelength
Injections

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Determination of pineal melatonin by precolumn derivatization reversed- phase high-performance liquid chromatography and its application to the study of circadian rhythm in rats and mice. / Hamase, Kenji; Tomita, Tatsunosuke; Kiyomizu, Ayako; Zaitsu, Kiyoshi.

In: Analytical Biochemistry, Vol. 279, No. 1, 01.03.2000, p. 106-110.

Research output: Contribution to journalArticle

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AB - Determination of minute amounts of endogenous melatonin in rat and mouse pineal gland was performed using an RP-HPLC system. Melatonin was separated following precolumn derivatization and determined with a fluorescence detector at the emission wavelength of 380 nm with the excitation at 245 nm. The calibration curve of melatonin constructed by adding known amounts of melatonin to the homogenates of mouse pineal gland was linear over the range of 1-500 fmol (injection amount/20 μl). The detection limit of added melatonin was 1 fmol (S/N = 5). Repeatability and day-to-day precision for the melatonin spiked sample of mouse pineal gland was 4.0 and 3.8% (RSD), respectively. Using the present method, circadian changes of melatonin content in rat (Wistar) and mouse (C3H) pineal gland were determined. In addition, a minute amount of melatonin in ddY mouse pineal gland was determined, because pineal melatonin of many inbred mouse strains has been reported to be lower than the detection limit. (C) 2000 Academic Press.

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