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 language | English |
|---|---|
| Pages (from-to) | 106-110 |
| Number of pages | 5 |
| Journal | Analytical Biochemistry |
| Volume | 279 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Mar 1 2000 |
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All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology
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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 journal › Article
}
TY - JOUR
T1 - 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
AU - Hamase, Kenji
AU - Tomita, Tatsunosuke
AU - Kiyomizu, Ayako
AU - Zaitsu, Kiyoshi
PY - 2000/3/1
Y1 - 2000/3/1
N2 - 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.
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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U2 - 10.1006/abio.1999.4435
DO - 10.1006/abio.1999.4435
M3 - Article
VL - 279
SP - 106
EP - 110
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 1
ER -