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<p>Acrylamide (AA) and 5-hydroxymethylfurfural (5-HMF) are heat-induced heterocyclic compounds with known carcinogenic and neurotoxic potential that are typically formed during thermal food processing. This study described the applicability of dispersive liquid–liquid microextraction (DLLME) method, coupled with high-performance liquid chromatography and UV detection (HPLC–UV), for the quantitative determination of AA and 5-HMF in Malaysian fried fish sausages (locally known as keropok lekor) samples. The separation was effected using a core–shell C18 column, isocratic elution with MeOH:H<sub>2</sub>O (10:90, v/v), resulting in a total run time of &lt; 10&#xa0;min. Detection wavelengths were set at 210&#xa0;nm for AA and 280&#xa0;nm for 5-HMF. The DLLME parameters were initially screened using a fractional factorial design to identify significant factors (sample volume, extractant volume, dispersant volume, and pH), followed by optimization using Box-Behnken design. The optimized DLLME conditions included 100 μL of chloroform (extractant), 500 μL of acetone (dispersant), and 5&#xa0;mL sample volume adjusted to pH 5. The DLLME-HPLC–UV method demonstrated good linearity across the studied ranges (R<sup>2</sup> = 0.9944 for AA and 0.9965 for 5-HMF), with detection limits (S/N = 3) of 10&#xa0;µg L<sup>−1</sup> for AA and 2&#xa0;µg L<sup>−1</sup> for 5-HMF. Intra- and inter-day precision showed relative standard deviations (RSDs) of ≤ 3.35% for retention times and peak areas. Recoveries of AA were 81.29–99.62% and 94.90–102.9% for 5-HMF. The proposed method exhibited greenness scores of 0.34 (AGREEprep) and 0.49 (AGREE). The validated approach was successfully applied to local fried fish sausage samples, where AA concentrations ranged from 586.5–1158&#xa0;µg&#xa0;kg<sup>−1</sup> and 5-HMF from 12.54–23.43&#xa0;mg&#xa0;kg<sup>−1</sup></p>

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Quantitative Determination of Acrylamide and 5-Hydroxymethylfurfural in Fried Fish Sausage Using An Improved Dispersive Liquid–liquid Microextraction and HPLC–UV Method

  • 期刊论文

  • 发表日期：

    2026年4月23日

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  • Song Peiyu,
  • Chen Minghong,
  • Tien Ping Lee,
  • Yin-Hui Leong,
  • Aliyu Ibrahim Kangiwa,
  • Musa I. Mohammed,
  • Yong Foo Wong

摘要

Acrylamide (AA) and 5-hydroxymethylfurfural (5-HMF) are heat-induced heterocyclic compounds with known carcinogenic and neurotoxic potential that are typically formed during thermal food processing. This study described the applicability of dispersive liquid–liquid microextraction (DLLME) method, coupled with high-performance liquid chromatography and UV detection (HPLC–UV), for the quantitative determination of AA and 5-HMF in Malaysian fried fish sausages (locally known as keropok lekor) samples. The separation was effected using a core–shell C18 column, isocratic elution with MeOH:H2O (10:90, v/v), resulting in a total run time of < 10 min. Detection wavelengths were set at 210 nm for AA and 280 nm for 5-HMF. The DLLME parameters were initially screened using a fractional factorial design to identify significant factors (sample volume, extractant volume, dispersant volume, and pH), followed by optimization using Box-Behnken design. The optimized DLLME conditions included 100 μL of chloroform (extractant), 500 μL of acetone (dispersant), and 5 mL sample volume adjusted to pH 5. The DLLME-HPLC–UV method demonstrated good linearity across the studied ranges (R2 = 0.9944 for AA and 0.9965 for 5-HMF), with detection limits (S/N = 3) of 10 µg L−1 for AA and 2 µg L−1 for 5-HMF. Intra- and inter-day precision showed relative standard deviations (RSDs) of ≤ 3.35% for retention times and peak areas. Recoveries of AA were 81.29–99.62% and 94.90–102.9% for 5-HMF. The proposed method exhibited greenness scores of 0.34 (AGREEprep) and 0.49 (AGREE). The validated approach was successfully applied to local fried fish sausage samples, where AA concentrations ranged from 586.5–1158 µg kg−1 and 5-HMF from 12.54–23.43 mg kg−1