Profiling of tryptophan-related plasma indoles in patients with carcinoid tumors by automated, on-line, solid-phase extraction and HPLC with fluorescence detection.
BACKGROUND
Profiling of the plasma indoles tryptophan, 5-hydroxytryptophan (5-HTP), serotonin, and 5-hydroxyindoleacetic acid (5-HIAA) is useful in the diagnosis and follow-up of patients with carcinoid tumors. We describe an automated method for the profiling of these indoles in protein-containing matrices as well as the plasma indole concentrations in healthy controls and patients with carcinoid tumors.


METHODS
Plasma, cerebrospinal fluid, and tissue homogenates were prepurified by automated on-line solid-phase extraction (SPE) in Hysphere Resin SH SPE cartridges containing strong hydrophobic polystyrene resin. Analytes were eluted from the SPE cartridge by column switching. Subsequent separation and detection were performed by reversed-phase HPLC combined with fluorometric detection in a total cycle time of 20 min. We obtained samples from 14 healthy controls and 17 patients with metastasized midgut carcinoid tumors for plasma indole analysis. In the patient group, urinary excretion of 5-HIAA and serotonin was compared with concentrations of plasma indoles.


RESULTS
Within- and between-series CVs for indoles in platelet-rich plasma were 0.6-6.2% and 3.7-12%, respectively. Results for platelet-rich plasma serotonin compared favorably with those obtained by single-component analysis. Plasma 5-HIAA, but not 5-HTP was detectable in 8 of 17 patients with carcinoid tumors. In the patient group, platelet-rich plasma total tryptophan correlated negatively with platelet-rich plasma serotonin (P = 0.021; r = -0.56), urinary 5-HIAA (P = 0.003; r = -0.68), and urinary serotonin (P <0.0001; r = -0.80).


CONCLUSIONS
The present chromatographic approach reduces analytical variation and time needed for analysis and gives more detailed information about metabolic deviations in indole metabolism than do manual, single-component analyses.
Kema
I P
IP
Department of Pathology and Laboratory Medicine, University Hospital Groningen, 9700 RB Groningen, The Netherlands. i.p.kema@lab.azg.nl
Meijer
W G
WG
Meiborg
G
G
Ooms
B
B
Willemse
P H
PH
de Vries
E G
EG
D006916:5-Hydroxytryptophan / Q000032:analysis / Q000097:blood; D000328:Adult; D000368:Aged; D001322:Autoanalysis; D002276:Carcinoid Tumor / Q000097:blood* / Q000737:chemistry; D002555:Cerebrospinal Fluid / Q000737:chemistry; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006801:Humans; D006897:Hydroxyindoleacetic Acid / Q000032:analysis / Q000097:blood; D007211:Indoles / Q000032:analysis / Q000097:blood*; D007414:Intestinal Neoplasms / Q000097:blood* / Q000737:chemistry; D008297:Male; D008875:Middle Aged; D011786:Quality Control; D015203:Reproducibility of Results; D012701:Serotonin / Q000032:analysis / Q000097:blood; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000032:analysis / Q000097:blood*; D014556:Urine / Q000737:chemistry
D007211:Indoles; D012701:Serotonin; D006897:Hydroxyindoleacetic Acid; D014364:Tryptophan; D006916:5-Hydroxytryptophan