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Alcohol biomarker analysis: simultaneous determination of 5-hydroxytryptophol glucuronide and 5-hydroxyindoleacetic acid by direct injection of urine using ultra-performance liquid chromatography-tandem mass spectrometry.
J Mass Spectrom. 2007 Jul; 42(7):940-9.JM

Abstract

A direct ultra-performance liquid chromatography-tandem mass spectrometry method (UPLC-MS/MS) for simultaneous measurement of urinary 5-hydroxytryptophol glucuronide (GTOL) and 5-hydroxyindoleacetic acid (5-HIAA) was developed. The GTOL/5-HIAA ratio is used as an alcohol biomarker with clinical and forensic applications. The method involved dilution of the urine sample with deuterated analogues (internal standards), reversed-phase chromatography with gradient elution, electrospray ionisation and monitoring of two product ions per analyte in selected reaction monitoring mode. The measuring ranges were 6.7-10 000 nmol/l for GTOL and 0.07-100 micromol/l for 5-HIAA. The intra- and inter-assay imprecision, expressed as the coefficient of variation, was below 7%. Influence from ion suppression was noted for both compounds but was compensated for by the use of co-eluting internal standards. The accuracy in analytical recovery of added substance to urine samples was 96 and 98%, respectively, for GTOL and 5-HIAA. Method comparison with GC-MS for GTOL in 25 authentic patient samples confirmed the accuracy of the method with a median ratio between methods (GC-MS to UPLC-MS/MS) of 1.14 (r(2) = 0.975). The difference is explained by the fact that the GC-MS method also measures unconjugated 5-hydroxytryptophol naturally present in urine. The comparison with data for 5-HIAA obtained by an HPLC method demonstrated a median ratio of 1.05 between the methods. The UPLC-MS/MS method was capable of measuring endogenous GTOL and 5-HIAA levels in urine, which agreed with the literature data. In conclusion, a fully validated and robust direct method for the routine measurement of urinary GTOL and 5-HIAA was developed.

Authors+Show Affiliations

Stephanson N
Department of Medicine, Division of Clinical Pharmacology, Karolinska Institute and University Hospital, SE-171 76 Stockholm, Sweden.
Helander A
No affiliation info available
Beck O
No affiliation info available

MeSH

Alcohol DrinkingBiomarkersChromatography, High Pressure LiquidGas Chromatography-Mass SpectrometryGlucuronidesHumansHydroxyindoleacetic AcidHydroxytryptopholIndicators and ReagentsReproducibility of ResultsTandem Mass Spectrometry

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17565712

Citation

Stephanson, Nikolai, et al. "Alcohol Biomarker Analysis: Simultaneous Determination of 5-hydroxytryptophol Glucuronide and 5-hydroxyindoleacetic Acid By Direct Injection of Urine Using Ultra-performance Liquid Chromatography-tandem Mass Spectrometry." Journal of Mass Spectrometry : JMS, vol. 42, no. 7, 2007, pp. 940-9.
Stephanson N, Helander A, Beck O. Alcohol biomarker analysis: simultaneous determination of 5-hydroxytryptophol glucuronide and 5-hydroxyindoleacetic acid by direct injection of urine using ultra-performance liquid chromatography-tandem mass spectrometry. J Mass Spectrom. 2007;42(7):940-9.
Stephanson, N., Helander, A., & Beck, O. (2007). Alcohol biomarker analysis: simultaneous determination of 5-hydroxytryptophol glucuronide and 5-hydroxyindoleacetic acid by direct injection of urine using ultra-performance liquid chromatography-tandem mass spectrometry. Journal of Mass Spectrometry : JMS, 42(7), 940-9.
Stephanson N, Helander A, Beck O. Alcohol Biomarker Analysis: Simultaneous Determination of 5-hydroxytryptophol Glucuronide and 5-hydroxyindoleacetic Acid By Direct Injection of Urine Using Ultra-performance Liquid Chromatography-tandem Mass Spectrometry. J Mass Spectrom. 2007;42(7):940-9. PubMed PMID: 17565712.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Alcohol biomarker analysis: simultaneous determination of 5-hydroxytryptophol glucuronide and 5-hydroxyindoleacetic acid by direct injection of urine using ultra-performance liquid chromatography-tandem mass spectrometry. AU - Stephanson,Nikolai, AU - Helander,Anders, AU - Beck,Olof, PY - 2007/6/15/pubmed PY - 2007/9/22/medline PY - 2007/6/15/entrez SP - 940 EP - 9 JF - Journal of mass spectrometry : JMS JO - J Mass Spectrom VL - 42 IS - 7 N2 - A direct ultra-performance liquid chromatography-tandem mass spectrometry method (UPLC-MS/MS) for simultaneous measurement of urinary 5-hydroxytryptophol glucuronide (GTOL) and 5-hydroxyindoleacetic acid (5-HIAA) was developed. The GTOL/5-HIAA ratio is used as an alcohol biomarker with clinical and forensic applications. The method involved dilution of the urine sample with deuterated analogues (internal standards), reversed-phase chromatography with gradient elution, electrospray ionisation and monitoring of two product ions per analyte in selected reaction monitoring mode. The measuring ranges were 6.7-10 000 nmol/l for GTOL and 0.07-100 micromol/l for 5-HIAA. The intra- and inter-assay imprecision, expressed as the coefficient of variation, was below 7%. Influence from ion suppression was noted for both compounds but was compensated for by the use of co-eluting internal standards. The accuracy in analytical recovery of added substance to urine samples was 96 and 98%, respectively, for GTOL and 5-HIAA. Method comparison with GC-MS for GTOL in 25 authentic patient samples confirmed the accuracy of the method with a median ratio between methods (GC-MS to UPLC-MS/MS) of 1.14 (r(2) = 0.975). The difference is explained by the fact that the GC-MS method also measures unconjugated 5-hydroxytryptophol naturally present in urine. The comparison with data for 5-HIAA obtained by an HPLC method demonstrated a median ratio of 1.05 between the methods. The UPLC-MS/MS method was capable of measuring endogenous GTOL and 5-HIAA levels in urine, which agreed with the literature data. In conclusion, a fully validated and robust direct method for the routine measurement of urinary GTOL and 5-HIAA was developed. SN - 1076-5174 UR - https://www.unboundmedicine.com/medline/citation/17565712/Alcohol_biomarker_analysis:_simultaneous_determination_of_5_hydroxytryptophol_glucuronide_and_5_hydroxyindoleacetic_acid_by_direct_injection_of_urine_using_ultra_performance_liquid_chromatography_tandem_mass_spectrometry_ L2 - https://doi.org/10.1002/jms.1231 DB - PRIME DP - Unbound Medicine ER -