Analysis of Acrylamide in Potato Chips by UHPLC–MS/MS

The Application Notebook

The Application Notebook, The Application Notebook-09-02-2018, Volume 36, Issue 9
Pages: 709

Acrylamide is viewed as a human health concern and found in certain foods after preparation or processing at high temperatures. World experts recommend reducing acrylamide in our diet and suggest long-term research studies to determine its potential risk. In this application we present an HPLC–MS/MS method for analysis of acrylamide in potato chips, using a simple sample preparation procedure.

Experimental Conditions

Stock solution of acrylamide (0.20 mg/mL) was prepared by dissolving 20 mg of the compounds in 100 mL of methanol. The standard working solutions were prepared by serial dilution sample; calibration levels were: 0.5–1.0–5.0–10–50–100 µg/L in methanol/water 50/50 and containing 100 µg/L of isotopic 13C-acrylamide. Samples were previously homogenized. Acrylamide extraction was as follows:

(1) Analytically weigh 1.0 ± 0.1 g crushed potato chips.

(2) Add 100 µL of 10 mg/L standard solution of 13C-acrylamide.

(3) Add 25 mL of 0.1% formic acid in methanol solution and mix in a shaker for 15 min.

(4) Centrifugation for 5 min (3000 rpm).

(5) Add 1 mL of extract to 1 mL of water.

(6) Condition C18 SPE tube (6 mL, 500 mg) with 2 mL methanol and 2 mL water/methanol 50/50: then, dry under vacuum.

(7) Apply 2 mL of extracted potato chip solution to SPE tube.

Acrylamide was separated on a PerkinElmer QSight® LX50 HPLC system using a UHPLC BEH C18 column (2.1 × 50 mm, 2.7 µm). Column temperature was set at 40 °C.

Figure 1: Linearity plot for standard solution over a range of 0.5-100 µg/L (internal standard).

Detection of acrylamide was carried out on a QSight triple quadrupole mass spectrometer equipped with an electrospray ionization source operating in positive ion mode and multiple reaction monitoring mode (MRM). Mass spectrometer conditions were as follows: ElectroSpray, 5000 V; Source Temp, 300 °C; HSID Temp, 275 °C; Drying Gas, 100 mL/min; Nebulizer Gas, 300 mL/min. Data acquisition and processing was performed using the Simplicity™ 3Q software.

Figure 2: MRM chromatogram of 0.5 and 5 µg/L acrylamide calibration solutions.


The selectivity of the method was evaluated by adding a known concentration of acrylamide equivalent to the limit of quantification to a matrix target (25 µg/kg). No matrix effect was observed by comparing solvent solution and extract solutions (the variation of the mean of the responses of 10 trials between both samples was less than 10%). Matrix concentration in sample extracts is very low (20 mg/mL).

Based on the calculated LOQ of 25 µg/kg, this method was in compliance with the technical requirements set by Regulation (EU) 2017/2158 (recovery 75-110%, RSD < Horwitz modified, LOQ ≤ 50 µg/kg). Very good recoveries were obtained (from 85 to 109%).


The QSight triple quad provides a robust platform for the analysis of trace acrylamide levels in compliance with European Regulations. This rapid, sensitive, and reproducible method can also be applied to other types of regulated matrices such as bread, biscuits, breakfast cereals, and more.

PerkinElmer, Inc.
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