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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.
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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/)

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