LC-MS/MS Analysis of Ethylene Glycol in Blood Using a Simple Derivatization Method

User Benefits

- A new quick and simple pretreatment allows for the measurement of ethylene glycol levels in blood by simultaneously removingproteins and derivatization. - Since it uses the same mobile phase as the “LC/MS/MS Rapid Toxicology Screening System”, it can be run on the same system with just a column change. - This method does not use expensive reagents and isotope-labeled internal standards, slowing for low-cost running.

Introduction

Ethylene glycol is an industrial product used in antifreeze and other applications. However, it is also a toxic substance that causes metabolic acidosis and renal failure when absorbed by the body. As a highly polar, low molecular weight molecule, ethylene glycol is only weakly retained on reversed-phase columns and is not readily detected by LC-MS analysis without some form of pretreatment. It is also highly hydrophilic, difficult to isolate from blood, and not easily measured by GC without pretreatment. These issues led to the development of a GC-MS method that detects phenylboronic derivatives of ethylene glycol produced by derivatization reactions in aqueous solutions. However, the phenylboronic acid used by this method can accumulate in GC inserts and columns, affecting subsequent measurements. This study investigated using a boronic acid derivatization reagent (2-bromopyridine-5-boronic acid [BPBA]) compatible with LC-MS analysis to develop a simple and rapid method for measuring ethylene glycol levels in blood. BPBA selectively reacts with the diol groups of ethylene glycol, forming a stable cyclic boronic ester. The nitrogen atom in this cyclic boronic ester improves its electrospray ionization (ESI) efficiency, whereas BPBA reacts quickly with ethylene glycol in aqueous samples. Taking advantage of these attributes, a simple and rapid pretreatment process was developed that performs deproteinization and derivatization simultaneously.

November 25, 2025 GMT