High-performance liquid chromatographic analysis of melatonin in human plasma and rabbit serum with on-line column enrichment.
This paper describes the development of a simple and sensitive analytical method for the quantification of melatonin in human plasma and rabbit serum, using standard analytical equipment and on-line column enrichment without prior extraction, clean-up or derivatization. The analytical procedure was found to be accurate, precise and linear. For human plasma, the accuracy was 101% (range 89-106%), and the mean precision was 5% (range 2-9%) for all concentrations (0, 2, 10, 50 and 200 ng/ml) tested (n=6). The accuracy in rabbit serum was 101% (range 90-112%), and the mean precision was 13% (range 8-19%) for all concentrations (0, 2, 10, 50, 200 and 500 ng/ml) tested (n=6). The retention time of melatonin was about 8 min and the total recoveries were found to be approximately 65 and 85%, respectively, for human plasma and rabbit serum. The limit of detection was found to be lower than 1 ng/ml for human plasma and around 2 ng/ml for rabbit serum. The method is, therefore, found to be suitable for melatonin bioavailability studies in rabbits and presumably also in humans.
The Royal Danish School of Pharmacy, Department of Pharmaceutics, Copenhagen.
E
E
Bechgaard
K
K
Lindhardt
L
L
Martinsen
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008550:Melatonin / Q000097:blood*; D011817:Rabbits; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
D008550:Melatonin