Detection of mutagenic activity in urine samples using a new concentration procedure

Area of Science:

• Toxicology
• Analytical Chemistry
• Pharmacology

Background:

• Cyclophosphamide (CP) is a widely used chemotherapeutic agent.
• Urinary metabolites of CP, such as nor-nitrogen mustard (NNM), can retain mutagenic properties.
• Accurate detection and concentration of these mutagens are crucial for toxicological assessment.

Purpose of the Study:

• To compare the efficacy of Polygosil C-18 and XAD-2 resins for concentrating mutagenic activity in rat urine.
• To determine the optimal resin for recovering cyclophosphamide and its primary urinary metabolite.

Main Methods:

• Urine samples from rats treated with cyclophosphamide were analyzed.
• Mutagenic activity was detected using Salmonella typhimurium tester strain TA1535.
• Separation and concentration of mutagens were performed using Polygosil C-18 and XAD-2 resins, followed by methanol elution.

Main Results:

• Polygosil C-18 resin demonstrated superior performance in concentrating mutagenic activity compared to XAD-2 resin.
• Nor-nitrogen mustard (NNM) was irreversibly bound to XAD-2 resin, preventing its recovery.
• NNM could be effectively recovered after elution with methanol from Polygosil C-18 resin.
• Experiments with varying doses of CP confirmed the better efficacy of Polygosil C-18.

Conclusions:

• Polygosil C-18 resin is the preferred method for concentrating cyclophosphamide and its mutagenic metabolites from rat urine.
• This improved concentration technique enhances the detection and toxicological evaluation of CP-induced mutagenicity.