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Measurement of plasma 5-hydroxyindole acetic acid by liquid chromatography tandem mass spectrometry--comparison with HPLC methodology.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Mar 01; 878(7-8):695-9.JC

Abstract

In patients with carcinoid disease, urinary concentration of the serotonin metabolite 5-hydroxyindole acetic acid (5-HIAA) is currently used to monitor disease progression or response to treatment as it is the metabolic end-product resulting from free and stored serotonin turnover. However, due to the undignified, cumbersome and error-prone nature of 24-h urine collections, there is constant pressure to replace them. It has been demonstrated using high performance liquid chromatography (HPLC) with fluorescence detection technology that plasma can achieve this, with the added advantage that it can be used for diagnostic purposes also. Here we describe a much simpler method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) that is twice as fast as a HPLC method currently in routine use. The sample preparation protocol requires 50 micorL of plasma and a simple protein precipitation step facilitated by acetonitrile. Chromatography was performed on a Phenomenex C18 Security Guard column coupled to a SIELC Primesep B reversed-phase, anion-exchange dual chemistry column and methanolic mobile phase gradient elution. Eluant was directly connected to a Waters Quattro Premier XE tandem mass spectrometer operating in positive ion mode. We detected multiple reaction monitoring transitions m/z 191.9>145.6 and 193.9>147.6 for 5-HIAA and d2-5-HIAA respectively, which co-eluted at 2.1 min. Ion suppression was negligible, recovery from spiked plasma was 103% (range 97-113%) and the method showed good linearity to 10,000 nmol/L (r(2)=0.999). Within-batch and between-batch imprecision was <10% and bias <15% at 3 concentrations, the limit of detection was 5 nmol/L and lower limit of quantitation 15 nmol/L. No interference was observed with l-tryptophan or 5-hydroxytryptamine. Comparison of LC-MS/MS and HPLC showed good agreement between the two methods but this LC-MS/MS assay displays several advantages; it requires 10-fold less sample, has a simpler extraction procedure and extended linearity, thus increasing laboratory throughput, lowering reagent costs and removing the need to dilute samples in patients with established carcinoid disease being monitored for therapeutic efficacy.

Authors+Show Affiliations

Department of Clinical Biochemistry, University Hospital of South Manchester, Southmoor Road, Wythenshawe, Manchester M23 9LT, UK. adrian.miller@nhs.netNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

20117968

Citation

Miller, Adrian G., et al. "Measurement of Plasma 5-hydroxyindole Acetic Acid By Liquid Chromatography Tandem Mass Spectrometry--comparison With HPLC Methodology." Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, vol. 878, no. 7-8, 2010, pp. 695-9.
Miller AG, Brown H, Degg T, et al. Measurement of plasma 5-hydroxyindole acetic acid by liquid chromatography tandem mass spectrometry--comparison with HPLC methodology. J Chromatogr B Analyt Technol Biomed Life Sci. 2010;878(7-8):695-9.
Miller, A. G., Brown, H., Degg, T., Allen, K., & Keevil, B. G. (2010). Measurement of plasma 5-hydroxyindole acetic acid by liquid chromatography tandem mass spectrometry--comparison with HPLC methodology. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 878(7-8), 695-9. https://doi.org/10.1016/j.jchromb.2010.01.010
Miller AG, et al. Measurement of Plasma 5-hydroxyindole Acetic Acid By Liquid Chromatography Tandem Mass Spectrometry--comparison With HPLC Methodology. J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Mar 1;878(7-8):695-9. PubMed PMID: 20117968.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Measurement of plasma 5-hydroxyindole acetic acid by liquid chromatography tandem mass spectrometry--comparison with HPLC methodology. AU - Miller,Adrian G, AU - Brown,Heather, AU - Degg,Tim, AU - Allen,Keith, AU - Keevil,Brian G, Y1 - 2010/01/18/ PY - 2009/08/04/received PY - 2009/12/18/revised PY - 2010/01/11/accepted PY - 2010/2/2/entrez PY - 2010/2/2/pubmed PY - 2010/6/16/medline SP - 695 EP - 9 JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences JO - J Chromatogr B Analyt Technol Biomed Life Sci VL - 878 IS - 7-8 N2 - In patients with carcinoid disease, urinary concentration of the serotonin metabolite 5-hydroxyindole acetic acid (5-HIAA) is currently used to monitor disease progression or response to treatment as it is the metabolic end-product resulting from free and stored serotonin turnover. However, due to the undignified, cumbersome and error-prone nature of 24-h urine collections, there is constant pressure to replace them. It has been demonstrated using high performance liquid chromatography (HPLC) with fluorescence detection technology that plasma can achieve this, with the added advantage that it can be used for diagnostic purposes also. Here we describe a much simpler method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) that is twice as fast as a HPLC method currently in routine use. The sample preparation protocol requires 50 micorL of plasma and a simple protein precipitation step facilitated by acetonitrile. Chromatography was performed on a Phenomenex C18 Security Guard column coupled to a SIELC Primesep B reversed-phase, anion-exchange dual chemistry column and methanolic mobile phase gradient elution. Eluant was directly connected to a Waters Quattro Premier XE tandem mass spectrometer operating in positive ion mode. We detected multiple reaction monitoring transitions m/z 191.9>145.6 and 193.9>147.6 for 5-HIAA and d2-5-HIAA respectively, which co-eluted at 2.1 min. Ion suppression was negligible, recovery from spiked plasma was 103% (range 97-113%) and the method showed good linearity to 10,000 nmol/L (r(2)=0.999). Within-batch and between-batch imprecision was <10% and bias <15% at 3 concentrations, the limit of detection was 5 nmol/L and lower limit of quantitation 15 nmol/L. No interference was observed with l-tryptophan or 5-hydroxytryptamine. Comparison of LC-MS/MS and HPLC showed good agreement between the two methods but this LC-MS/MS assay displays several advantages; it requires 10-fold less sample, has a simpler extraction procedure and extended linearity, thus increasing laboratory throughput, lowering reagent costs and removing the need to dilute samples in patients with established carcinoid disease being monitored for therapeutic efficacy. SN - 1873-376X UR - https://www.unboundmedicine.com/medline/citation/20117968/Measurement_of_plasma_5_hydroxyindole_acetic_acid_by_liquid_chromatography_tandem_mass_spectrometry__comparison_with_HPLC_methodology_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1570-0232(10)00022-X DB - PRIME DP - Unbound Medicine ER -