Determination of serotonin and bufotenin as their dansyl derivatives.
A method is described which permits the determination of serotonin and bufotenin in the same tissue sample. It comprises the following steps: (a) tissue extraction with acetone-0.1 M hydrochloric acid (19:1); (b) reaction of the tissue extract with dansyl (Dns) chloride; (c) pre-separation of O-Dns-bufotenin from O,N-bis-Dns-serotonin and other Dns-amides on a small silica gel column (this step is dispensable if only serotonin or bufotonin is being determined); (d) TLC separation of O-Dns-bufotenin and O,N-bis-Dns-serotonin from other Dns derivatives; (e) quantitative evaluation of the separated compounds by fluorimetry for O-Dns-bufotenin and by fluorimetry or mass spectrometry for the serotonin derivative. The photometer response is linear within the range 0.1-300 nmole. With the mass spectrometric method, 2 pmole of O,N-bis-Dns-serotonin could be determined with a standard deivation of +/-9%. The recovery of the amines from tissue was better than 85%. Reserpine treatment of toads caused a concomitant decrease in serotonin and bufotenin in the brain, but not in the skin of the animals. Repletion of bufotenin in the brain occurs at a higher rate than the repletion of the serotonin pool.
D000818:Animals; D001001:Anura; D001923:Brain Chemistry; D002027:Bufotenin / Q000032:analysis*; D002855:Chromatography, Thin Layer; D003619:Dansyl Compounds / Q000032:analysis*; D013058:Mass Spectrometry; D008722:Methods; D012110:Reserpine / Q000032:analysis; D012701:Serotonin / Q000031:analogs & derivatives* / Q000032:analysis*; D012867:Skin / Q000032:analysis; D013050:Spectrometry, Fluorescence
D003619:Dansyl Compounds; D002027:Bufotenin; D012701:Serotonin; D012110:Reserpine