Chromatographia

, Volume 75, Issue 21–22, pp 1257–1269 | Cite as

The Application and Validation of HybridSPE-Precipitation Cartridge Technology for the Rapid Clean-up of Serum Matrices (from Phospholipids) for the Clinical Analysis of Serotonin, Dopamine and Melatonin

  • Merisa Moriarty
  • Aoife Lee
  • Brendan O’Connell
  • Mary Lehane
  • Helen Keeley
  • Ambrose Furey
Original

Abstract

Phospholipids have been shown to cause matrix effects particularly in liquid chromatography–mass spectrometry (LC–MS) analysis of small molecules. This results in suppression of the analyte signal. This study provides a versatile validated method for the analysis of serotonin in serum along with dopamine and melatonin using LC–MS/MS. It utilises HybridSPE-Precipitation cartridges for the clean-up of serum samples. This technology involves a simple protein precipitation step together with a fast and robust SPE method that is designed to remove phospholipids. Serotonin and dopamine are major neurotransmitters in the brain which affect various functions both in the brain and in the rest of the body. Melatonin plays an important role in the regulation of circadian sleep–wake cycle. Good linear calibrations were obtained for the multiplex assay of analytes in serum samples (0.021–3.268 μmol L−1; R2 = 0.9983–0.9993). Acceptable intra- and inter-day repeatability was achieved for all analytes in serum. Excellent limits of detection (LOD) and limits of quantitation (LOQ) were achieved with LODs of 3.2–23.5 nmol L−1 and the LOQs of 15.4–70.5 nmol L−1 for these analytes in serum. The sample clean-up procedure that was developed provided efficient recovery and reproducibility while also decreasing preparation time and solvent use. A sample storage protocol was established, this was achieved by investigation of sample stability under different storage conditions. Evaluation of matrix effects was also carried out and the influence of ion suppression on analytical results reported. This clean-up protocol was then applied to the analysis of clinical serum samples.

Keywords

LC–MS/MS HybridSPE-Precipitation Serotonin Dopamine Melatonin Ion suppression Serum 

Supplementary material

10337_2012_2330_MOESM1_ESM.ppt (87 kb)
Online resource 1: Example of a post-column infusion experiment for the evaluation of ion suppression in serum matrix for serotonin (a) and dopamine (b). Shaded areas correspond to elution times of analytes. Analyte standards (2.84, 3.26 and 2.15 μmol L−1 for serotonin, dopamine and melatonin) were infused at a rate of 10 μL min−1. This experiment was repeat with different serum samples (n = 5) (PPT 87 kb)
10337_2012_2330_MOESM2_ESM.ppt (70 kb)
Online resource 2: Dilution study for the assessment of matrix ion suppression on the target compound, serotonin, comparing various dilutions of serum. With no ion suppression effects from serum matrices at zero dilution, a 1 in 2 dilution of the samples should given exactly a 50 % response. This study determined that a 1 in 5 dilution is necessary to eliminate completely the influence of matrix ion suppression effects (PPT 69 kb)

References

  1. 1.
    Muller C, Schafer P, Stortzel M, Vogt S, Weinmann W (2002) Ion suppression effects in liquid chromatography-electrospray-ionisation transport-region collision induced dissociation mass spectrometry with different serum extraction methods for systematic toxicological analysis with mass spectra libraries. J Chromatogr B 773(1):47–52CrossRefGoogle Scholar
  2. 2.
    Ismaiel OA, Halquist MS, Elmamly MY, Shalaby A, Karnes HT (2007) Monitoring phospholipids for assessment of matrix effects in a liquid chromatography–tandem mass spectrometry method for hydrocodone and pseudoephedrine in human plasma. J Chromatogr B 859:84–93CrossRefGoogle Scholar
  3. 3.
    Tsao C-W, Lin Y-S, Chen C–C, Bai C-H, Wu S-R (2006) Cytokines and serotonin transporter in patients with major depression. Prog Neuro-Psychoph 30(5):899–905CrossRefGoogle Scholar
  4. 4.
    Ishida J, Takada M, Yamaguchi M (1997) 3,4-Dimethoxybenzylamine as a sensitive pre-column fluorescence derivatization reagent for the determination of serotonin in human platelet-poor plasma. J Chromatogr B 692(1):31–36CrossRefGoogle Scholar
  5. 5.
    Nutt D, Argyropoulos S, Hood S, Potokar J (2006) Generalized anxiety disorder: a comorbid disease. Eur Neuropsychopharm 16:S109–S118CrossRefGoogle Scholar
  6. 6.
    Mitani H, Shirayama Y, Yamada T, Kawahara R (2006) Plasma levels of homovanillic acid, 5-hydroxyindoleacetic acid and cortisol, and serotonin turnover in depressed patients. Prog Neuro- Psychoph 30(3):531–534CrossRefGoogle Scholar
  7. 7.
    Kema IP, de Vries EGE, Muskiet FAJ (2000) Clinical chemistry of serotonin and metabolites. J Chromatogr Biomed 747(1–2):33–48CrossRefGoogle Scholar
  8. 8.
    Martínez A, Knappskog PM, Haavik J (2001) A structural approach into human tryptophan hydroxylase and its implications for the regulation of serotonin biosynthesis. Curr Med Chem 8:1077–1091Google Scholar
  9. 9.
    Peterson ZD, Lee ML, Graves SW (2004) Determination of serotonin and its precursors in human plasma by capillary electrophoresis-electrospray ionization-time-of-flight mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 810(1):101–110Google Scholar
  10. 10.
    Eriksson K, Ostin A, Levin JO (2003) Quantification of melatonin in human saliva by liquid chromatography–tandem mass spectrometry using stable isotope dilution. J Chromatogr B Anal Technol Biomed Life Sci 794(1):115–123CrossRefGoogle Scholar
  11. 11.
    Yang SM, Zheng XH, Xu Y, Zhou X (2002) Rapid determination of serum melatonin by ESI–MS–MS with direct sample injection. J Pharm Biomed Anal 30(3):781–790CrossRefGoogle Scholar
  12. 12.
    Carta M, Fadda F, Stancampiano R (2006) Tryptophan-deficient diet increases the neurochemical and behavioral response to amphetamine. Brain Res 1094(1):86–91CrossRefGoogle Scholar
  13. 13.
    Antkiewicz-Michaluk L, Romanska I, Papla I, Michaluk J, Bakalarz M, Vetulani J, Krygowska-Wajs A, Szczudlik A (2000) Neurochemical changes induced by acute and chronic administration of 1,2,3,4-tetrahydroisoquinoline and salsolinol in dopaminergic structures of rat brain. Neuroscience 96(1):59–64CrossRefGoogle Scholar
  14. 14.
    Lam KSL, Aman MG, Arnold LE (2006) Neurochemical correlates of autistic disorder: a review of the literature. Res Dev Disabil 27(3):254–289CrossRefGoogle Scholar
  15. 15.
    Stephanson N, Dahl H, Helander A, Beck O (2005) Determination of urinary 5-hydroxytryptophol glucuronide by liquid chromatography–mass spectrometry. J Chromatogr Biomed 816(1–2):107–112Google Scholar
  16. 16.
    Ma N, L-w Tan, Wang Q, Z-X Li, L-J Li (2007) Lower levels of whole blood serotonin in obsessive-compulsive disorder and in schizophrenia with obsessive-compulsive symptoms. Psychiatry Res 150(1):61–69CrossRefGoogle Scholar
  17. 17.
    Monaghan PJ, Brown HA, Houghton LA, Keevil BG (2009) Measurement of serotonin in platelet depleted plasma by liquid chromatography tandem mass spectrometry. J Chromatogr B 877(22):2163–2167CrossRefGoogle Scholar
  18. 18.
    Miller AG, Brown H, Degg T, Allen K, Keevil BG (2010) Measurement of plasma 5-hydroxyindole acetic acid by liquid chromatography tandem mass spectrometry—comparison with HPLC methodology. J Chromatogr B 878(7–8):695–699CrossRefGoogle Scholar
  19. 19.
    Umeda S, Stagliano GW, Borenstein MR, Raffa RB (2005) A reverse-phase HPLC and fluorescence detection method for measurement of 5-hydroxytryptamine (serotonin) in Planaria. J Pharmzcol Toxicol 51(1):73–76CrossRefGoogle Scholar
  20. 20.
    Lang WS, Masucci JA, Caldwell GW, Hageman W, Hall J, Jones WJ, Rafferty BM (2004) Liquid chromatographic and tandem mass spectrometric assay for evaluation of in vivo inhibition of rat brain monoamine oxidases (MAO) A and B following a single dose of MAO inhibitors: application of biomarkers in drug discovery. Anal Biochem 333(1):79–87CrossRefGoogle Scholar
  21. 21.
    Johnson RD, Lewis RJ, Canfield DV, Blank CL (2004) Accurate assignment of ethanol origin in postmortem urine: liquid chromatographic-mass spectrometric determination of serotonin metabolites. J Chromatogr B 805(2):223–234CrossRefGoogle Scholar
  22. 22.
    de Jong WHA, Graham KS, de Vries EGE, Kema IP (2008) Urinary 5-HIAA measurement using automated on-line solid-phase extraction–high-performance liquid chromatography–tandem mass spectrometry. J Chromatogr B 868(1–2):28–33CrossRefGoogle Scholar
  23. 23.
    Supelco Hybrid SPE-Precipitation Technology. BrochureGoogle Scholar
  24. 24.
    Pucci V, Di Palma S, Alfieri A, Bonelli F, Monteagudo E (2009) A novel strategy for reducing phospholipids-based matrix effect in LC–ESI–MS bioanalysis by means of HybridSPE. J Pharm Biomed Anal 50:867–871CrossRefGoogle Scholar
  25. 25.
    Ardjomand-Woelkart K, Kollroser M, Li L, Derendorf H, Butterweck V, Bauer R (2011) Development and Validation of a LC–MS/MS method based on a new 96-well HybridSPE-precipitation technique for quantification of a CYP450 substrates/metabolites in rat plasma. Anal Bioanal Chem 400:2371–2381CrossRefGoogle Scholar
  26. 26.
    Jiang H, Zhang Y, Ida M, LaFayette A, Fast DM (2011) Determination of Carboplatin in human plasma using HybridSPE-precipitation along with liquid chromatography–tandem mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 879:2162–2170CrossRefGoogle Scholar
  27. 27.
    Choi BK, Hercules DM, Gusev AI (2001) LC–MS/MS signal suppression effects in the analysis of pesticides in complex environmental matrices. Fresenius J Anal Chem 369(3–4):370–377CrossRefGoogle Scholar
  28. 28.
    Antignac J-P, de Wasch K, Monteau F, De Brabander H, Andre F, Le Bizec B (2005) The ion suppression phenomenon in liquid chromatography–mass spectrometry and its consequences in the field of residue analysis. Anal Chim Acta 529(1–2):129–136CrossRefGoogle Scholar
  29. 29.
    Benijts T, Dams R, Lambert W, De Leenheer A (2004) Countering matrix effects in environmental liquid chromatography–electrospray ionization tandem mass spectrometry water analysis for endocrine disrupting chemicals. J Chromatogr A 1029(1–2):153–159Google Scholar
  30. 30.
    Kall MA, Fu I, Dige T, Vallano P, Woolf E, Jorgensen M (2007) Development and validation of a selective and sensitive bioanalytical procedure for the quantitative determination of gaboxadol in human plasma employing mixed mode solid phase extraction and hydrophilic interaction liquid chromatography with tandem mass spectroscopic detection. J Chromatogr B 858(1–2):168–176CrossRefGoogle Scholar
  31. 31.
    Commission decision of 12 August 2002 implementing council directive 96/23/EC concerning the performance of analytical methods and the interpretation of results (notified under document number C (2002) 3044) (Text with EEA relevance) (2002/657/EC). L 221/8-36 EN Official Journal of the European Communities 1782002Google Scholar
  32. 32.
    International Conference on Harmonisation of technical requirements for registration of pharmaceutical for human use: Validation of analytical procedures: Text and Methodology Q2 (R1) Current Step 4 version, Parent Guideline dated 27 October 1994 (Complementary Guidelines on Methodology dated 6 November 1996 incorporated in November 2005)Google Scholar
  33. 33.
    Moriarty M, Lee A, O’Connell B, Kelleher A, Keeley H, Furey A (2011) Development of an LC–MS/MS method for the analysis of serotonin and related compounds in urine and the identification of a potential biomarker for attention deficit hyperactivity/hyperkinetic disorder. Anal Bioanal Chem 401(8):2481–2493CrossRefGoogle Scholar
  34. 34.
    Ismaiel OA, Halquist MS, Elmamly MY, Shalaby A, Thomas Karnes H (2008) Monitoring phospholipids for assessment of ion enhancement and ion suppression in ESI and APCI LC/MS/MS for chlorpheniramine in human plasma and the importance of multiple source matrix effect evaluations. J Chromatogr B 875(2):333–343CrossRefGoogle Scholar
  35. 35.
    Hows MEP, Lacroix L, Heidbreder C, Organ AJ, Shah AJ (2004) High-performance liquid chromatography/tandem mass spectrometric assay for the simultaneous measurement of dopamine, norepinephrine, 5-hydroxytryptamine and cocaine in biological samples. J Neurosci Methods 138(1–2):123–132CrossRefGoogle Scholar
  36. 36.
    Semak I, Korik E, Naumova M, Wortsman J, Slominski A (2004) Serotonin metabolism in rat skin: characterization by liquid chromatography–mass spectrometry. Arch Biochem Biophys 421(1):61–66CrossRefGoogle Scholar
  37. 37.
    Koppisetti G, Siriki A, Sukala K, Subbaraju GV (2005) Estimation of L-5-hydroxytryptophan in rat serum and Griffonia seed extracts by liquid chromatography-mass spectrometry. Anal Chim Acta 549(1–2):129–133CrossRefGoogle Scholar
  38. 38.
    Villagrasa M, Guillamon M, Eljarrat E, Barcelo D (2007) Matrix effect in liquid chromatography–electrospray ionization mass spectrometry analysis of benzoxazinoid derivatives in plant material. J Chromatogr A 1157(1–2):108–114Google Scholar
  39. 39.
    Saracino MA, Gerra G, Somaini L, Colombati M, Raggi MA (2010) Chromatographic analysis of serotonin, 5-hydroxyindolacetic acid and homovanillic acid in dried blood spots and platelet poor and rich plasma samples. J Chromatogr A 1217(29):4808–4814CrossRefGoogle Scholar
  40. 40.
    Laganà A, Marino A, Fago G, Pardo-Martinez B, Bizzarri M (1995) Sensitive assay for melatonin in human serum by liquid chromatography. Anal Chim Acta 316(3):377–385CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Merisa Moriarty
    • 1
  • Aoife Lee
    • 2
  • Brendan O’Connell
    • 2
  • Mary Lehane
    • 1
  • Helen Keeley
    • 3
  • Ambrose Furey
    • 4
  1. 1.Department of Chemistry, PROTEOBIO (Mass Spectrometry Centre for Proteomic and Biotoxin Research)Cork Institute of TechnologyCorkIreland
  2. 2.Department of Biological SciencesCork Institute of TechnologyBishopstownIreland
  3. 3.Child and Adolescent Mental Health Services, Health Service Executive, SouthNorth Cork Area and the National Suicide Research FoundationCorkIreland
  4. 4.Team Elucidate and PROTEOBIO (Mass Spectrometry Centre for Proteomic and Biotoxin Research) research groupsCork Institute of TechnologyCorkIreland

Personalised recommendations