PDF herunterladen
CC BY-NC-ND 4.0 · J Lab Physicians
DOI: 10.1055/s-0043-1774378
Original Article

Validated Ultraviolet High-Performance Liquid Chromatography Method for Post-mortem 5-Hydroxy-indoleacetic Acid Measurement in Human Cerebrospinal Fluid

Divya Bhushan
1   Department Forensic Medicine and Toxicology, All India Institute of Medical Sciences Bhopal, Bhopal, Madhya Pradesh, India
,
Jayanthi Yadav
1   Department Forensic Medicine and Toxicology, All India Institute of Medical Sciences Bhopal, Bhopal, Madhya Pradesh, India
,
Arneet Arora
1   Department Forensic Medicine and Toxicology, All India Institute of Medical Sciences Bhopal, Bhopal, Madhya Pradesh, India
,
Dhruv Agrawal 
2   Department of Medical Laboratory Sciences, LPU, Phagwara, Punjab, India
,
Achala Apte
3   Department of Biological Sciences, IISER, Bhopal, Madhya Pradesh, India
› Institutsangaben Funding The author(s) disclose receipt of the following financial support for this article's research, authorship, and publication: ICMR MD/MS financial thesis support.
› Weitere Informationen
   Lizenzen und Reprints

Abstract

Objective The objective of this study was to develop an ultraviolet high-performance liquid chromatography (UV-HPLC) method for the measurement of 5-hydroxyindoleacetic acid (5-HIAA) in human cerebrospinal fluid (CSF) as a potential biomarker for neurological and psychiatric illnesses, including depressive disorders with suicidal behavior.

Methods The study utilized CSF samples from individuals brought for medicolegal autopsy. The 5-HIAA concentration was measured using a UV-HPLC method with three mobile-phase solvents. The most effective mobile phase solvent was then used to measure 5-HIAA in the CSF samples.

Materials The materials used in the study included CSF samples obtained from individuals brought for medico-legal autopsy, UV-HPLC equipment, and mobile phase solvents, including 5-hydroxyindole-3-acetic acid (Merck/sigma), acetonitrile, concentrated formic acid, concentrated acetic acid, methanol, and phosphoric acid.

Statistical Analysis This was done using R Studio (version 4.2.0).

Results The study found that the UV-HPLC method utilizing formic acid (0.05–0.1%): acetonitrile in a 1:9 as mobile phase was the most effective for measuring 5-HIAA in human CSF. The method exhibited excellent linearity, accuracy, and precision.

Conclusion The study concludes that the developed UV-HPLC method is reliable and effective for measuring 5-HIAA in human CSF. Measuring 5-HIAA levels in CSF can serve as a potential biomarker for neurological and psychiatric illnesses, including depressive disorders with suicidal behavior. This method is promising for clinical and forensic practice to diagnose suicidal cases. Further research is needed to determine the clinical significance of these findings and the potential for broader application in psychiatry. This article helps to give a practical, cost-effective methodology to detect CSF 5-HIAA.

Keywords

5-hydroxyindoleacetic acid - cerebrospinal fluid - high-performance liquid chromatography (HPLC) - suicidal

Authors' Contributions

D.B. helped in inception of idea and carrying out the work and manuscript preparation. J.Y. was responsible for inception of idea, guidance, and supervision and writing of the manuscript. A.A. helped in supervision and facilitation for work and editing of the manuscript and carrying out the laboratory work and analyzing data. D.A. was involved in guidance and supervision in analyzing the data.


The study was approved by Institutional human ethics committee of AIIMS Bhopal vide LOP letter no 2020/PG/Jan/09 dated Nov 21, 2020.




Publikationsverlauf

Eingereicht: 14. März 2023

Angenommen: 01. August 2023

Artikel online veröffentlicht:
05. September 2023

© 2023. The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India

 

  • References

  • 1 Vernau W, Vernau KA, Sue Bailey C. Cerebrospinal Fluid. Clinical Biochemistry of Domestic Animals. . Elsevier Publisher; 2008: 769-819
    Google Scholar
  • 2 Shaywitz BA, Cohen DJ, Bowers MB. Reduced cerebrospinal fluid 5-hydroxy indole acetic acid and homovanillic acid in children with epilepsy. Neurology 1975; 25 (01) 72 DOI: 10.1212/WNL.25.1.72.
    CrossrefPubMedGoogle Scholar
  • 3 Mathew SJ, Coplan JD, Smith ELP. et al. Cerebrospinal fluid concentrations of biogenic amines and corticotropin-releasing factor in adolescent non-human primates as a function of the timing of adverse early rearing. Stress 2002; 5 (03) 185-193
    CrossrefPubMedGoogle Scholar
  • 4 Lenchner JR, Santos C. Biochemistry, 5 Hydroxyindoleacetic Acid. 2022 May 8. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; ; 2022 Jan–. PMID: 31869148
    Google Scholar
  • 5 Menon KN, Steer DL, Short M, Petratos S, Smith I, Bernard CCA. A novel unbiased proteomic approach to detect the reactivity of cerebrospinal fluid in neurological diseases. Mol Cell Proteomics 2011; 10 (06) 000042
    CrossrefPubMedGoogle Scholar
  • 6 Carlborg A, Jokinen J, Nordström AL, Jönsson EG, Nordström P. CSF 5-HIAA, attempted suicide and suicide risk in schizophrenia spectrum psychosis. Schizophr Res 2009; 112 (1-3): 80-85
    CrossrefPubMedGoogle Scholar
  • 7 Jayamohananan H, Manoj Kumar MK, Aneesh TP. 5-HIAA as a potential biological marker for neurological and psychiatric disorders. Adv Pharm Bull 2019; 9 (03) 374-381
    CrossrefPubMedGoogle Scholar
  • 8 Correlation Between Salivary, Platelet and Central Serotonin Levels in Children | Canadian Journal of Neurological Sciences | Cambridge Core [Internet]. [cited 2022 Nov 3]. Accessed August 18, 2023 at: https://www.cambridge.org/core/journals/canadian-journal-of-neurological-sciences/article/correlation-between-salivary-platelet-and-central-serotonin-levels-in-children/98A7C394671DBF747CF97E88B944C6D2
    Google Scholar
  • 9 Collins CM, Kloek J, Elliott JM. Parallel changes in serotonin levels in brain and blood following acute administration of MDMA. J Psychopharmacol 2013; 27 (01) 109-112
    CrossrefPubMedGoogle Scholar
  • 10 Hrishi AP, Sethuraman M. Cerebrospinal fluid (CSF) analysis and interpretation in neurocritical care for acute neurological conditions. Indian J Crit Care Med 2019; 23 (Suppl. 02) S115-S119
    CrossrefPubMedGoogle Scholar
  • 11 Takada A, Shimizu F, Masuda J. Measurement of plasma tryptophan metabolites: clinical and experimental application for depression and stress states assessment. Melatonin Molecular Biology Clinical and Pharmaceutical Approaches 2018; x: x
    Google Scholar
  • 12 Fujino K, Yoshitake T, Kehr J, Nohta H, Yamaguchi M. Simultaneous determination of 5-hydroxyindoles and catechols by high-performance liquid chromatography with fluorescence detection following derivatization with benzylamine and 1,2-diphenylethylenediamine. J Chromatogr A 2003; 1012 (02) 169-177
    CrossrefPubMedGoogle Scholar
  • 13 Vaarmann A, Kask A, Mäeorg U. Novel and sensitive high-performance liquid chromatographic method based on electrochemical coulometric array detection for simultaneous determination of catecholamines, kynurenine and indole derivatives of tryptophan. J Chromatogr B Analyt Technol Biomed Life Sci 2002; 769 (01) 145-153
    CrossrefPubMedGoogle Scholar
  • 14 Miller AG, Brown H, Degg T, Allen K, Keevil BG. 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): 695-699
    CrossrefPubMedGoogle Scholar
  • 15 González RR, Fernández RF, Vidal JL, Frenich AG, Pérez ML. Development and validation of an ultra-high performance liquid chromatography-tandem mass-spectrometry (UHPLC-MS/MS) method for the simultaneous determination of neurotransmitters in rat brain samples. J Neurosci Methods 2011; 198 (02) 187-194
    CrossrefPubMedGoogle Scholar
  • 16 Koel M, Nebinger P. Determination of urinary 5-hydroxy-3-indoleacetic acid by an automated HPLC method. Biomed Chromatogr 1989; 3 (03) 114-117
    CrossrefPubMedGoogle Scholar
  • 17 Chapter 3: Investigating Proteins – Chemistry [Internet]. [cited 2023 Feb 16]. Accessed August 18, 2023 at: https://wou.edu/chemistry/courses/online-chemistry-textbooks/ch450-and-ch451-biochemistry-defining-life-at-the-molecular-level/chapter-3-investigating-proteins/
    Google Scholar