Quantitation of melatonin and n-acetylserotonin in human plasma by nanoflow LC-MS/MS and electrospray LC-MS/MS
Melatonin (MEL) and its chemical precursor N‐acetylserotonin (NAS) are believed to be potential biomarkers for sleep‐related disorders. Measurement of these compounds, however, has proven to be difficult due to their low circulating levels, especially that of NAS. Few methods offer the sensitivity,...
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| Vydané v: | Journal of mass spectrometry. Ročník 47; číslo 3; s. 277 - 285 |
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| Hlavní autori: | , , , , |
| Médium: | Journal Article |
| Jazyk: | English |
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Chichester, UK
John Wiley & Sons, Ltd
01.03.2012
Wiley |
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| ISSN: | 1076-5174, 1096-9888, 1096-9888 |
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| Abstract | Melatonin (MEL) and its chemical precursor N‐acetylserotonin (NAS) are believed to be potential biomarkers for sleep‐related disorders. Measurement of these compounds, however, has proven to be difficult due to their low circulating levels, especially that of NAS. Few methods offer the sensitivity, specificity and dynamic range needed to monitor MEL and its precursors and metabolites in small blood samples, such as those obtained from pediatric patients. In support of our ongoing study to determine the safety, tolerability and PK dosing strategies for MEL in treating insomnia in children with autism spectrum disorder, two highly sensitive LC‐MS/MS assays were developed for the quantitation of MEL and precursor NAS at pg/mL levels in small volumes of human plasma. A validated electrospray ionization (ESI) method was used to quantitate high levels of MEL in PK studies, and a validated nanospray (nESI) method was developed for quantitation of MEL and NAS at endogenous levels. In both assays, plasma samples were processed by centrifugal membrane dialysis after addition of stable isotopic internal standards, and the components were separated by either conventional LC using a Waters SymmetryShield RP18 column (2.1 × 100 mm, 3.5 µm) or on a polyimide‐coated, fused‐silica capillary self‐packed with 17 cm AquaC18 (3 µm, 125 Å). Quantitation was done using the SRM transitions m/z 233 → 174 and m/z 219 → 160 for MEL and NAS, respectively. The analytical response ratio versus concentration curves were linear for MEL (nanoflow LC: 11.7–1165 pg/mL, LC: 1165–116500 pg/mL) and for NAS (nanoflow LC: 11.0–1095 pg/mL). Copyright © 2012 John Wiley & Sons, Ltd. |
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| AbstractList | Melatonin (MEL) and its chemical precursor N-acetylserotonin (NAS) are believed to be potential biomarkers for sleep-related disorders. Measurement of these compounds, however, has proven to be difficult due to their low circulating levels, especially that of NAS. Few methods offer the sensitivity, specificity and dynamic range needed to monitor MEL and its precursors and metabolites in small blood samples, such as those obtained from pediatric patients. In support of our ongoing study to determine the safety, tolerability, and PK dosing strategies for MEL in treating insomnia in children with autism spectrum disorder, two highly sensitive LC-MS/MS assays were developed for the quantitation of MEL and precursor NAS at pg/mL levels in small volumes of human plasma. A validated electrospray ionization (ESI) method was used to quantitate high levels of MEL in PK studies and a validated nanospray (nESI) method was developed for quantitation of MEL and NAS at endogenous levels. In both assays plasma samples were processed by centrifugal membrane dialysis after addition of stable isotopic internal standards, and the components were separated by either conventional LC using a Waters SymmetryShield RP18 column (2.1×100 mm, 3.5 μm) or on a polyimide-coated, fused-silica capillary self-packed with 17 cm AquaC18 (3 μm, 125 Å). Quantitation was done using the SRM transitions m/z 233→174 and m/z 219→160 for MEL and NAS, respectively. The analytical response ratio vs. concentration curves were linear for MEL (nanoflow LC: 11.7–1165 pg/mL, LC: 1165–116500 pg/mL) and for NAS (nanoflow LC: 11.0–1095 pg/mL). Melatonin (MEL) and its chemical precursor N-acetylserotonin (NAS) are believed to be potential biomarkers for sleep-related disorders. Measurement of these compounds, however, has proven to be difficult due to their low circulating levels, especially that of NAS. Few methods offer the sensitivity, specificity and dynamic range needed to monitor MEL and its precursors and metabolites in small blood samples, such as those obtained from pediatric patients. In support of our ongoing study to determine the safety, tolerability and PK dosing strategies for MEL in treating insomnia in children with autism spectrum disorder, two highly sensitive LC-MS/MS assays were developed for the quantitation of MEL and precursor NAS at pg/mL levels in small volumes of human plasma. A validated electrospray ionization (ESI) method was used to quantitate high levels of MEL in PK studies, and a validated nanospray (nESI) method was developed for quantitation of MEL and NAS at endogenous levels. In both assays, plasma samples were processed by centrifugal membrane dialysis after addition of stable isotopic internal standards, and the components were separated by either conventional LC using a Waters SymmetryShield RP18 column (2.1 × 100 mm, 3.5 µm) or on a polyimide-coated, fused-silica capillary self-packed with 17 cm AquaC18 (3 µm, 125 Å). Quantitation was done using the SRM transitions m/z 233 → 174 and m/z 219 → 160 for MEL and NAS, respectively. The analytical response ratio versus concentration curves were linear for MEL (nanoflow LC: 11.7-1165 pg/mL, LC: 1165-116,500 pg/mL) and for NAS (nanoflow LC: 11.0-1095 pg/mL). Melatonin (MEL) and its chemical precursor N‐acetylserotonin (NAS) are believed to be potential biomarkers for sleep‐related disorders. Measurement of these compounds, however, has proven to be difficult due to their low circulating levels, especially that of NAS. Few methods offer the sensitivity, specificity and dynamic range needed to monitor MEL and its precursors and metabolites in small blood samples, such as those obtained from pediatric patients. In support of our ongoing study to determine the safety, tolerability and PK dosing strategies for MEL in treating insomnia in children with autism spectrum disorder, two highly sensitive LC‐MS/MS assays were developed for the quantitation of MEL and precursor NAS at pg/mL levels in small volumes of human plasma. A validated electrospray ionization (ESI) method was used to quantitate high levels of MEL in PK studies, and a validated nanospray (nESI) method was developed for quantitation of MEL and NAS at endogenous levels. In both assays, plasma samples were processed by centrifugal membrane dialysis after addition of stable isotopic internal standards, and the components were separated by either conventional LC using a Waters SymmetryShield RP18 column (2.1 × 100 mm, 3.5 µm) or on a polyimide‐coated, fused‐silica capillary self‐packed with 17 cm AquaC18 (3 µm, 125 Å). Quantitation was done using the SRM transitions m/z 233 → 174 and m/z 219 → 160 for MEL and NAS, respectively. The analytical response ratio versus concentration curves were linear for MEL (nanoflow LC: 11.7–1165 pg/mL, LC: 1165–116500 pg/mL) and for NAS (nanoflow LC: 11.0–1095 pg/mL). Copyright © 2012 John Wiley & Sons, Ltd. Melatonin (MEL) and its chemical precursor N-acetylserotonin (NAS) are believed to be potential biomarkers for sleep-related disorders. Measurement of these compounds, however, has proven to be difficult due to their low circulating levels, especially that of NAS. Few methods offer the sensitivity, specificity and dynamic range needed to monitor MEL and its precursors and metabolites in small blood samples, such as those obtained from pediatric patients. In support of our ongoing study to determine the safety, tolerability and PK dosing strategies for MEL in treating insomnia in children with autism spectrum disorder, two highly sensitive LC-MS/MS assays were developed for the quantitation of MEL and precursor NAS at pg/mL levels in small volumes of human plasma. A validated electrospray ionization (ESI) method was used to quantitate high levels of MEL in PK studies, and a validated nanospray (nESI) method was developed for quantitation of MEL and NAS at endogenous levels. In both assays, plasma samples were processed by centrifugal membrane dialysis after addition of stable isotopic internal standards, and the components were separated by either conventional LC using a Waters SymmetryShield RP18 column (2.1 × 100 mm, 3.5 µm) or on a polyimide-coated, fused-silica capillary self-packed with 17 cm AquaC18 (3 µm, 125 Å). Quantitation was done using the SRM transitions m/z 233 → 174 and m/z 219 → 160 for MEL and NAS, respectively. The analytical response ratio versus concentration curves were linear for MEL (nanoflow LC: 11.7-1165 pg/mL, LC: 1165-116,500 pg/mL) and for NAS (nanoflow LC: 11.0-1095 pg/mL).Melatonin (MEL) and its chemical precursor N-acetylserotonin (NAS) are believed to be potential biomarkers for sleep-related disorders. Measurement of these compounds, however, has proven to be difficult due to their low circulating levels, especially that of NAS. Few methods offer the sensitivity, specificity and dynamic range needed to monitor MEL and its precursors and metabolites in small blood samples, such as those obtained from pediatric patients. In support of our ongoing study to determine the safety, tolerability and PK dosing strategies for MEL in treating insomnia in children with autism spectrum disorder, two highly sensitive LC-MS/MS assays were developed for the quantitation of MEL and precursor NAS at pg/mL levels in small volumes of human plasma. A validated electrospray ionization (ESI) method was used to quantitate high levels of MEL in PK studies, and a validated nanospray (nESI) method was developed for quantitation of MEL and NAS at endogenous levels. In both assays, plasma samples were processed by centrifugal membrane dialysis after addition of stable isotopic internal standards, and the components were separated by either conventional LC using a Waters SymmetryShield RP18 column (2.1 × 100 mm, 3.5 µm) or on a polyimide-coated, fused-silica capillary self-packed with 17 cm AquaC18 (3 µm, 125 Å). Quantitation was done using the SRM transitions m/z 233 → 174 and m/z 219 → 160 for MEL and NAS, respectively. The analytical response ratio versus concentration curves were linear for MEL (nanoflow LC: 11.7-1165 pg/mL, LC: 1165-116,500 pg/mL) and for NAS (nanoflow LC: 11.0-1095 pg/mL). |
| Author | Carter, Melissa D. Wade Calcutt, M. Malow, Beth A. Rose, Kristie L. Hachey, David L. |
| AuthorAffiliation | Vanderbilt University, Department of Neurology, Nashville, TN 37232, USA Vanderbilt University, Department of Biochemistry, Nashville, TN 37235, USA Vanderbilt University, Department of Pharmacology, Nashville, TN 37232, USA |
| AuthorAffiliation_xml | – name: Vanderbilt University, Department of Neurology, Nashville, TN 37232, USA – name: Vanderbilt University, Department of Biochemistry, Nashville, TN 37235, USA – name: Vanderbilt University, Department of Pharmacology, Nashville, TN 37232, USA |
| Author_xml | – sequence: 1 givenname: Melissa D. surname: Carter fullname: Carter, Melissa D. organization: Department of Biochemistry, Vanderbilt University, TN, 37235, Nashville, USA – sequence: 2 givenname: M. surname: Wade Calcutt fullname: Wade Calcutt, M. organization: Department of Biochemistry, Vanderbilt University, TN, 37235, Nashville, USA – sequence: 3 givenname: Beth A. surname: Malow fullname: Malow, Beth A. organization: Department of Neurology, Vanderbilt University, TN, 37232, Nashville, USA – sequence: 4 givenname: Kristie L. surname: Rose fullname: Rose, Kristie L. organization: Department of Biochemistry, Vanderbilt University, TN, 37235, Nashville, USA – sequence: 5 givenname: David L. surname: Hachey fullname: Hachey, David L. email: david.l.hachey@vanderbilt.edu (D. L. Hachey), david.l.hachey@vanderbilt.edu organization: Department of Biochemistry, Vanderbilt University, 37235, Nashville, TN, USA |
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| Keywords | Biological fluid Tandem mass spectrometry Melatonin Chemical analysis Aminoacid derivative hormone HPLC chromatography Tryptamine derivatives N-acetylserotonin Developmental disorder Electrospray Blood Blood plasma Clinical biology Biosynthesis precursor autism spectrum disorder Child Quantitative analysis Human Healthy subject Coupled method Oral administration Patient Endogenous substance Autism Nanoflow LC-MS/MS Indole derivatives Pharmacokinetics |
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| References_xml | – reference: E. Gilad, N. Zisapel. High-affinity binding of melatonin to hemoglobin. Biochem Mol Med 1995, 56, 115. – reference: G. Kulman, et al. Evidence of pineal endocrine hypofunction in autistic children. Neuro Endocrinol Lett 2000, 21, 31. – reference: K. S. Reddy. Cytogenetic abnormalities and fragile-X syndrome in Autism Spectrum Disorder. BMC Med Genet 2005, 6, 3. – reference: J. B. Shear, et al. Determination of fluorogen-labeled neurotransmitters at the zeptomole level using two photon excited fluorescence with capillary electrophoresis. Anal. Chem. 1998, 70, 3470. – reference: D. A. Rossignol, R. E. Frye. Melatonin in autism spectrum disorders: a systematic review and meta-analysis. Dev Med Child Neurol 2011, 53, 783. – reference: M. Wilhelmsen, et al. Analgesic effects of melatonin: a review of current evidence from experimental and clinical studies. J Pineal Res 2011. – reference: G. A. Bubenik, S. J. Konturek. Melatonin and aging: prospects for human treatment. J. Physiol. Pharmacol. 2011, 62, 13. – reference: E. A. de Almeida, et al. Measurement of melatonin in body fluids: standards, protocols and procedures. Childs Nerv Syst 2011, 27, 879. – reference: G. Simonin, et al. Determination of melatonin in biological fluids in the presence of the melatonin agonist S 20098: comparison of immunological techniques and GC-MS methods. J. Pharm. Biomed. Anal. 1999, 21, 591. – reference: B. A. Malow, K. W. Adkins, S. G. McGrew, L. Wang, S. E. Goldman, D. Fawkes, C. Burnette. Melatonin for sleep in children with autism: A controlled trial examining dose, tolerability, and outcomes. J. Autism Dev. Disord. 2011 Dec. 10 (Epub ahead of print). – reference: R. M. Leu, et al. Relation of melatonin to sleep architecture in children with autism. J Autism Dev Disord 2011, 41, 427. – reference: I. Nir, et al. Brief report: circadian melatonin, thyroid-stimulating hormone, prolactin, and cortisol levels in serum of young adults with autism. J Autism Dev Disord 1995, 25, 641. – reference: T. Wang, et al. Effects of melatonin on orphanin FQ/nociceptin-induced hyperalgesia in mice. Brain Res 2006, 1085, 43. – reference: J. Melke, et al. Abnormal melatonin synthesis in autism spectrum disorders. Mol Psychiatry 2008, 13, 90. – reference: V. Rizzo, et al. Determination of free and total (free plus protein-bound) melatonin in plasma and cerebrospinal fluid by high-performance liquid chromatography with fluorescence detection. J Chromatogr B Analyt Technol Biomed Life Sci 2002, 774, 17. – reference: G. Chen, et al. Melatonin in Chinese medicinal herbs. Life Sci. 2003, 73, 19. – reference: A. Cavallo, W. A. Ritschel. Pharmacokinetics of melatonin in human sexual maturation. J. Clin. Endocrinol. Metab. 1996, 81, 1882. – reference: A. Ulugol, et al. Antihyperalgesic, but not antiallodynic, effect of melatonin in nerve-injured neuropathic mice: possible involvements of the L-arginine-NO pathway and opioid system. Life Sci. 2006, 78, 1592. – reference: G. M. Anderson. Genetics of childhood disorders: XLV. Autism, part 4: serotonin in autism. J Am Acad Child Adolesc Psychiatry 2002, 41, 1513. – reference: S. Tordjman, et al. Nocturnal excretion of 6-sulphatoxymelatonin in children and adolescents with autistic disorder. Biol. Psychiatry 2005, 57, 134. – reference: B. C. Koch, et al. Circadian sleep-wake rhythm disturbances in end-stage renal disease. Nat. Rev. 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Autism, part 4: serotonin in autism publication-title: J Am Acad Child Adolesc Psychiatry – volume: 1085 start-page: 43 year: 2006 article-title: Effects of melatonin on orphanin FQ/nociceptin‐induced hyperalgesia in mice publication-title: Brain Res – volume: 774 start-page: 17 year: 2002 article-title: Determination of free and total (free plus protein‐bound) melatonin in plasma and cerebrospinal fluid by high‐performance liquid chromatography with fluorescence detection publication-title: J Chromatogr B Analyt Technol Biomed Life Sci – volume: 27 start-page: 879 year: 2011 article-title: Measurement of melatonin in body fluids: standards, protocols and procedures publication-title: Childs Nerv Syst – volume: 21 start-page: 591 year: 1999 article-title: Determination of melatonin in biological fluids in the presence of the melatonin agonist S 20098: comparison of immunological techniques and GC‐MS methods publication-title: J. Pharm. Biomed. Anal. – volume: 56 start-page: 115 year: 1995 article-title: High‐affinity binding of melatonin to hemoglobin publication-title: Biochem Mol Med – year: 2011 article-title: Analgesic effects of melatonin: a review of current evidence from experimental and clinical studies publication-title: J Pineal Res – volume: 81 start-page: 1882 year: 1996 article-title: Pharmacokinetics of melatonin in human sexual maturation publication-title: J. Clin. Endocrinol. Metab. – ident: e_1_2_6_6_1 doi: 10.1111/j.1469-8749.2011.03980.x – volume: 70 start-page: 3470 year: 1998 ident: e_1_2_6_16_1 article-title: Determination of fluorogen‐labeled neurotransmitters at the zeptomole level using two photon excited fluorescence with capillary electrophoresis publication-title: Anal. Chem. doi: 10.1021/ac980296f – ident: e_1_2_6_20_1 doi: 10.1006/bmme.1995.1066 – ident: e_1_2_6_18_1 doi: 10.1016/S0731-7085(99)00150-8 – ident: e_1_2_6_10_1 doi: 10.1016/j.biopsych.2004.11.003 – ident: e_1_2_6_4_1 doi: 10.1016/j.lfs.2005.07.002 – ident: e_1_2_6_21_1 doi: 10.1210/jc.81.5.1882 – ident: e_1_2_6_15_1 doi: 10.1038/nrneph.2009.88 – ident: e_1_2_6_12_1 doi: 10.1097/00004583-200212000-00025 – year: 2011 ident: e_1_2_6_3_1 article-title: Analgesic effects of melatonin: a review of current evidence from experimental and clinical studies publication-title: J Pineal Res doi: 10.1111/j.1600-079X.2011.00895.x – ident: e_1_2_6_9_1 doi: 10.1007/BF02178193 – ident: e_1_2_6_19_1 doi: 10.1016/S1570-0232(02)00168-X – ident: e_1_2_6_11_1 doi: 10.1007/s10803-010-1072-1 – ident: e_1_2_6_14_1 doi: 10.1007/s00381-010-1278-8 – volume: 62 start-page: 13 year: 2011 ident: e_1_2_6_2_1 article-title: Melatonin and aging: prospects for human treatment publication-title: J. Physiol. Pharmacol. – ident: e_1_2_6_13_1 doi: 10.1186/1471-2350-6-3 – ident: e_1_2_6_17_1 doi: 10.1016/S0024-3205(03)00252-2 – ident: e_1_2_6_8_1 doi: 10.1038/sj.mp.4002016 – volume: 21 start-page: 31 year: 2000 ident: e_1_2_6_7_1 article-title: Evidence of pineal endocrine hypofunction in autistic children publication-title: Neuro Endocrinol Lett – year: 2011 ident: e_1_2_6_22_1 article-title: Melatonin for sleep in children with autism: A controlled trial examining dose, tolerability, and outcomes publication-title: J. Autism Dev. Disord. – ident: e_1_2_6_5_1 doi: 10.1016/j.brainres.2006.02.006 |
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| SubjectTerms | Adult Analysis autism spectrum disorder Biological and medical sciences Child Child clinical studies Child, Preschool Chromatography, Liquid - methods Developmental disorders Female General pharmacology Humans Infantile autism Investigative techniques, diagnostic techniques (general aspects) Male Medical sciences Melatonin Melatonin - blood Melatonin - pharmacokinetics Miscellaneous. Technology N-acetylserotonin Nanoflow LC-MS/MS Nanotechnology Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Pharmacology. Drug treatments Psychology. Psychoanalysis. Psychiatry Psychopathology. Psychiatry Reproducibility of Results Serotonin - analogs & derivatives Serotonin - blood Serotonin - pharmacokinetics Spectrometry, Mass, Electrospray Ionization - methods Tandem Mass Spectrometry - methods |
| Title | Quantitation of melatonin and n-acetylserotonin in human plasma by nanoflow LC-MS/MS and electrospray LC-MS/MS |
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