# An Automated Micro Solid-Phase Extraction (μSPE) Liquid Chromatography-Mass Spectrometry Method for Cyclophosphamide and Iphosphamide: Biological Monitoring in Antineoplastic Drug (AD) Occupational Exposure

## TL;DR

This paper introduces a fast and automated method to detect antineoplastic drugs in urine, improving safety monitoring for healthcare workers.

## Contribution

A novel µSPE-UHPLC-MS/MS method for detecting cyclophosphamide and iphosphamide in urine with high sensitivity and low solvent use.

## Key findings

- The µSPE extraction achieved 77–79% recovery with less than 1.5 mL solvent per 1 mL urine.
- The UHPLC-MS/MS method achieved LOQs below 10 pg/mL for cyclophosphamide and iphosphamide.
- Urinary contamination levels in hospital workers ranged from 27 to 182 pg/mL.

## Abstract

Despite the considerable steps taken in the last decade in the context of antineoplastic drug (AD) handling procedures, their mutagenic effect still poses a threat to healthcare personnel actively involved in compounding and administration units. Biological monitoring procedures usually require large volumes of sample and extraction solvents, or do not provide adequate sensitivity. It is here proposed a fast and automated method to evaluate the urinary levels of cyclophosphamide and iphosphamide, composed of a miniaturized solid phase extraction (µSPE) followed by ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis. The extraction procedure, developed through design of experiments (DoE) on the ePrep One Workstation, required a total time of 9.5 min per sample, with recoveries of 77–79% and a solvent consumption lower than 1.5 mL per 1 mL of urine sample. Thanks to the UHPLC-MS/MS method, the limits of quantification (LOQ) obtained were lower than 10 pg/mL. The analytical procedure was successfully applied to 23 urine samples from compounding wards of four Italian hospitals, which resulted in contaminations between 27 and 182 pg/mL.