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- Römsing, Susanne, et al.
(författare)
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Determination of melatonin in human plasma with solid-phase extraction, high-performance liquid chromatography and fluorescence detection
- 2003
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Ingår i: Scandinavian Journal of Clinical and Laboratory Investigation. - 0036-5513 .- 1502-7686. ; 63:1, s. 81-88
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Tidskriftsartikel (refereegranskat)abstract
- A new bioanalytical method for the determination of melatonin in plasma with high-performance liquid chromatography (HPLC) and fluorescence detection preceded by solid-phase extraction has been developed and validated. Melatonin was extracted from 3 mL plasma using a Waters Oasis HLB solid-phase extraction cartridge and the elute was evaporated to dryness and dissolved in 200 microl mobile phase; acetonitrile-phosphate buffer, 0.01 M pH 7.2 (25:75, v/v). 125 microL was injected into the HPLC system and separation was carried out on a Waters SymmetryShield RP18 column 5 microm (250 x 4.6 mm). Excitation and emission wavelengths were set to 285 nm and 345 nm, respectively. The HPLC system was able to separate melatonin and internal standard (5-fluorotryptamine) from other endogenous indole compounds such as serotonin and tryptophan. Determination down to 0.10 nmol/L was possible, with an intra-assay precision of about 13%. Melatonin was stable in plasma for at least 30 days at about 23 degrees C.
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| 3. |
- Römsing, Susanne, et al.
(författare)
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Determination of melatonin in saliva using solid-phase extraction, high-performance liquid chromatography and fluorescence detection
- 2006
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Ingår i: Scandinavian Journal of Clinical and Laboratory Investigation. - : Informa UK Limited. - 0036-5513 .- 1502-7686. ; 66:3, s. 181-190
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Tidskriftsartikel (refereegranskat)abstract
- A sensitive bioanalytical method for the determination of melatonin in saliva by solid-phase extraction (SPE), high-performance liquid chromatography (HPLC) and fluorescence detection has been developed and validated. Saliva was collected with a Salivette((R)) sampling device (Sarstedt) and a mixed-mode SPE column was used for the extraction of melatonin and internal standard (N-acetyl-6-methoxytryptamine) from the saliva. Chromatographic separation was performed using a HyPurity C18 LC column (150x2.1 mm) with mobile phase acetonitrile-ammonium hydrogen carbonate buffer, 0.015 M, pH 6.8 (23:77, v/v). Excitation and emission wavelengths were set to 285 nm and 345 nm, respectively. The within-day precision for the method at 50 pmol/L was 7.9% and the between-day precision was 10.5%. The limit of quantification was 50 pmol/L.
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