determination of Acrylamide by liquid chromatography

1. Reza Joia

2. BACKGROUND
2
Acrylamide is a known carcinogenic
product that has been found among the
substances such as potato chips which to
be processed under the heat-treatment. In
order to extract amounts of acrylamide
from fried chips in market, an ultrasound-
assisted liquid– liquid extraction (UA-
LLE) technique is proposed. The UA-
LLE coupled LLE and ultrasonication
in a single step
Reza Joia 12/25/2023

3. Materials and Methods
• The chemicals used in the study included high-purity acrylamide, methanol,
acetic acid, acetonitrile, and acetone, all of analytical grade, along with
Carrez I and Carrez II solutions obtained from reputable sources. Distilled,
deionized, and filtered water was employed consistently.
• A stock solution of acrylamide at 1 mg/mL was prepared using distilled water.
Calibration standards for the study were derived by diluting this stock
solution to concentrations of 0.5, 1.0, 3.0, 5.0, and 8.0 µg/mL with distilled
water. Carrez I and Carrez II solutions were also prepared for use in the
experimental procedure.
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Chemical and solutions:

4. INSTRUMENTS
• Acrylamide quantification was carried out using an Agilent
1100 HPLC system with a quaternary pump and vacuum
degasser. A C18 column (15 × 4.6 mm × 5µm) from Agilent
was used for chromatographic separations. Confirmatory
analysis of blank samples was performed on the same
instrument with a diode array detector (DAD). In routine
food sample analyses, the mobile phase (acidic water)
flowed at a rate of 1.0 mL/min, and acrylamide absorbance
was measured at 222 nm while monitoring the peak spectra
within the 190–400 nm range using a spectrophotometer.
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5. SAMPLE PREPARATION
1.Weighing: Weighing 1 gram of crushed potato chips to initiate the
sample preparation.
2.Spiking: Adding different levels of acrylamide to the samples for
recovery calculations. This step is crucial for determining the
accuracy of the analytical method.
3.Centrifugation: Centrifuging the mixture at 10°C and 10,000 rpm to
separate solid residues from the supernatant, allowing for the
purification of the acetone phase.
4.LC Column Injection: Injecting 20 µL of the prepared solution into
the Liquid Chromatography (LC) column for quantification. This step
is vital for the actual measurement of acrylamide levels in the sample.
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6. Results and Discussion
• In this study, the US-LLE process was optimized to
maximize recovery. Various parameters, including the
choice and volume of the extraction solvent, sonication
time, volume of the re-dissolving agent, and pH, were
optimized using a one-parameter-at-a-time method. The
efficiency of optimization was assessed by calculating the
recovery of acrylamide.
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7. OPTIMIZATION OF EXTRACTION PROCESS
1.The study aimed to optimize the extraction of acrylamide from potato chips, considering
parameters such as the choice of solvent, extraction method (sonication or vortexing),
and the addition of Carrez reagents.
2.Sonication was found to be more effective than vortex-assisted extraction in speeding up
the process. Therefore, an ultrasonic-assisted liquid-liquid extraction (US-LLE) method
was employed.
3.Acetone was chosen as the extraction solvent due to its high solubility for acrylamide and
compatibility with the chip matrix. The addition of Carrez solutions helped purify the
extract and prevent acrylamide loss during evaporation.
4.The study investigated the impact of various factors on sonication efficiency, including the
volume of extraction solvent (10 mL was optimal), sonication time (10 minutes proved
sufficient), and water volume for reconstitution (4 mL was adequate).
5.The pH of water used for acrylamide extraction was also studied, with an acidic pH of 3
yielding the highest recovery. The results suggested that acrylamide is released
effectively from water-soluble components of the chip matrix at this pH.
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8. Slide Title
Product A Product B
• Feature 1
• Feature 2
• Feature 3
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9. Continue
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10. Slide Title
Product A
• Feature 1
• Feature 2
• Feature 3
Product B
• Feature 1
• Feature 2
• Feature 3
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11. CONCLUSION
• Selection of Acetone as Solvent
• Ultrasonic-Assisted Liquid-Liquid Extraction (US-LLE):
• Ultrasonic-Assisted Liquid-Liquid Extraction (US-LLE):
• Optimal pH for Water Extraction:
• Use of Carrez Solutions
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12. THANKYOUFOR YOURATTENTION
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