# Quantitative Measurement of Hexoses by Betaine Aldehyde Derivatisation

## TL;DR

This paper introduces a new method for measuring hexoses like glucose using betaine aldehyde derivatisation and MALDI IMS, with applications in tissue analysis.

## Contribution

The study presents a novel biomimetic calibration approach for quantitative hexose analysis using MALDI IMS.

## Key findings

- The method successfully distinguishes between different hexoses in tissue samples.
- Optimal betaine aldehyde layers and matrix deposition techniques were identified for accurate calibration.
- Hexose levels varied significantly between brain, liver, kidney, and spinal cord tissues in control and morphine-addicted animals.

## Abstract

Hexoses, particularly glucose, are one of the most essential molecules for sustaining life; therefore, reliable methods for their analysis are very important. In our study, we present a qualitative and quantitative approach for analysing hexoses using MALDI IMS (Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging) with betaine aldehyde derivatisation and a CHCA (α-Cyano-4-hydroxycinnamic acid) matrix in positive ionisation mode. In this study, we demonstrated betaine aldehyde derivatisation of glucose from dried droplets and explored the analysis of hexoses in brain and liver tissue slices. We assessed whether our method could distinguish between mannose, galactose, glucose, and fructose and optimised the preparation of a biomimetic calibration curve using stable-isotope labelled glucose for hexose analysis. For this purpose, we investigated the number of betaine aldehyde layers required to obtain a proper calibration curve; examined whether changes in the spray nozzle position during CHCA matrix deposition could facilitate analysis and investigated how storage conditions influenced the calibration curve analysis. Finally, we optimised the technique for liver and brain analysis and assessed variations in hexose levels between brain, liver, kidney, and spinal cord tissues from control and morphine-addicted animals. We hope that our biomimetic approach to creating the calibration curve will be helpful for quantitative analysis and aid in developing various quantitative methods for assessing endogenous substances.