Quantitative Analysis of Acrylamide in Peanut Butter using LC Triple Quadrupole Mass Spectrometry

Importance of the Topic

Acrylamide is a probable carcinogen formed in carbohydrate‐rich foods during high‐temperature processing. Monitoring its levels in widely consumed products such as peanut butter is crucial for food safety, regulatory compliance, and public health risk assessment.

Objectives and Study Overview

This study aims to develop a rapid and straightforward sample preparation method compatible with a sensitive LC–MS/MS assay for quantifying acrylamide in peanut butter. It compares two calibration approaches—standard addition with isotopic internal standard (ISTD) and reversed ISTD calibration—to account for matrix effects and endogenous analyte levels.

Methodology and Instrumentation

Sample Preparation:

• One gram of peanut butter mixed with d3‐acrylamide ISTD (50 ng/g).
• Defatting step using hexane, followed by extraction with water and acetonitrile.
• Dispersive QuEChERS cleanup using AOAC kit salts and sorbents without solvent evaporation.
• Direct transfer of extract to autosampler vial.

LC–MS/MS Method:

• Agilent 1290 II UHPLC with Poroshell EC‐C18 column (3 × 150 mm, 2.7 µm).
• Isocratic elution (2 % acetonitrile with 0.1 % formic acid) at 0.25 mL/min, retaining acrylamide at 3.34 min.
• Flushing gradient to remove matrix residues and re‐equilibration for consistent retention times.

Instrumentation

• Agilent 1290 II UHPLC system.
• Agilent 6470 triple quadrupole mass spectrometer with AJS electrospray source.
• Bond Elut QuEChERS Extraction and AOAC Dispersive SPE kits (Agilent p/n 5982-5850, 5982-5022).

Main Results and Discussion

Calibration Performance:

• Standard Addition ISTD: Linear range 5–2000 ng/g (r2 > 0.9994), accuracy 87.3–107.9 %, RSD < 6.3 %.
• Reversed ISTD: Range 0.1–200 ng/g (r2 > 0.9994), accuracy 96.9–103.4 %, RSD < 6.15 %.
• Endogenous acrylamide estimated at 25.6 ng/g (standard addition) and 29.2 ng/g (reversed ISTD).

Recovery and Reproducibility:

• Sample recovery for d3‐acrylamide spiked pre- and post-prep: 65 %.
• Overall method precision (three independent preparations with triplicate LC–MS/MS): RSD < 4 %.

Method Suitability:

• Effective fat removal by hexane and matrix cleanup by dispersive SPE.
• Small injection volume and isocratic conditions minimize solvent‐related effects.
• Fast turnaround with simplified workflow.

Benefits and Practical Applications

The described procedure delivers a rapid, accurate, and reproducible way to quantify trace acrylamide in peanut butter. It reduces sample preparation time, avoids complex SPE evaporation steps, and achieves regulatory‐level sensitivity, making it suitable for routine QA/QC laboratories and food safety monitoring.

Future Trends and Applications

Advances may include automation of QuEChERS workflows, integration with high-throughput LC–MS platforms, and expansion to other complex matrices. Emerging ionization techniques and high-resolution mass spectrometry could further enhance sensitivity and selectivity for acrylamide and related contaminants.

Conclusion

This work presents a streamlined QuEChERS–LC–MS/MS method for acrylamide in peanut butter with excellent linearity, accuracy, and precision. Both calibration strategies effectively address matrix interferences, and the protocol supports rapid, high‐throughput analysis in food safety applications.

Reference

1. Yang Y, Li G, Chambers T. Quantitative Analysis of Acrylamide in Peanut Butter using LC Triple Quadrupole Mass Spectrometry. ASMS 2020 Poster Reprint WP 175.
2. Al-Taher F. Analysis of Acrylamide in French Fries using Agilent Bond Elut QuEChERS AOAC kit and LC–MS/MS. Agilent Application Note 5990-5940EN.
3. Mastovska K, Lehotay SJ. Rapid Sample Preparation Method for LC–MS/MS or GC–MS Analysis of Acrylamide in Various Food Matrices. J Agric Food Chem. 2006;54(19):7001–7008.