Detection Of Acrylamide Research Articles

An ultrasensitive liquid crystal aptasensing chip assisted by three-way junction DNA pockets for acrylamide detection in food samples

Acrylamide, an unsaturated amide found in heat-processed foods, poses serious risks to human health due to its neurotoxicity, carcinogenicity, and genotoxicity. This highlights the importance of quantitative determination of acrylamide in foods and the environments to ensure public health safety. Therefore, there is an urgent need for simple, rapid, and highly sensitive methods to accurately quantify acrylamide. In the present study, a user-friendly aptasensor was designed to quantify ultra-low levels of acrylamide in nuts for the first time. This innovative approach utilizes chemical engineering of a glass slide as a portable sensing platform, which incorporates liquid crystal (LC) molecules and a three-way junction (TWJ) DNA pocket. The immobilized TWJ pocket can disrupt the vertical alignment of LCs, turning the dark polarized background of the aptasensor to a colorful state. The binding of the specific aptamer to acrylamide disrupts the TWJ structure, enabling the LCs to return to their homotropic alignment. This structural change restores the dark polarized view of the sensing platform. The TWJ-engineered LC aptasensor effectively detects ultra-low concentrations of acrylamide in the range of 0.0005 to 50 fmol/L, with a detection limit of 0.106 amol/L. The aptasensor was successfully applied to real roasted nut samples, including peanut, almond, pistachio, and hazelnut, achieving recovery values ranging from 96.84 % to 99.61 %. With its simplicity, portability, ease of use, and cost-effectiveness, this aptasensor is a powerful sensing device for food safety monitoring.

Assessment of Acrylamide Levels by Advanced Molecularly Imprinted Polymer-Imprinted Surface Plasmon Resonance (SPR) Sensor Technology and Sensory Quality in Homemade Fried Potatoes.

This study evaluated acrylamide (AA) levels and various quality parameters in homemade fried potatoes prepared in different sizes by integrating principles from the Slow Food Movement with advanced sensor technology. To this aim, a surface plasmon resonance (SPR) sensor based on a molecularly imprinted polymer (MIP) was first developed for the determination of AA in homemade fried potatoes at low levels, and the AA levels in the samples were established. First of all, monolayer formation of allyl mercaptane on the SPR chip surface was carried out to form double bonds that could polymerize on the chip surface. AA-imprinted SPR chip surfaces modified with allyl mercaptane were prepared via UV polymerization using ethylene glycol dimethacrylate (EGDMA) as a cross-linker, N,N'-azobisisobutyronitrile (AIBN) as an initiator, and methacryloylamidoglutamicacid (MAGA) as a monomer. The prepared AA-imprinted and nonimprinted surfaces were characterized by atomic force microscopy (AFM) and Fourier transform infrared (FTIR) spectroscopy methods. The SPR sensor indicated linearity in the range of 1.0 × 10-9-5.0 × 10-8 M with a detection limit (LOD) of 3.0 × 10-10 M in homemade fried potatoes, and the SPR sensor demonstrated high selectivity and repeatability in terms of AA detection. Additionally, the highest AA level was observed in the potato sample belonging to the T1 group, at 15.37 nM (p < 0.05), and a strong and positive correlation was found between AA levels and sensory parameters, the a* value, the ΔE value, and the browning index (BI) (p < 0.05).

Dual emission BSA-capped bimetallic nanoclusters for detection of acrylamide in certain food matrices based on thiol-ene Michael addition click reaction

Dual-emissive gold and silver nanoclusters stabilized by bovine serum albumin (BSA@Au-Ag NCs) were engineered for the precise and sensitive estimation of acrylamide (ACM). The probe demonstrated emissions at 630 nm and 405 nm upon excitation at 320 nm. Decreasing the fluorescence of the probe at 630 nm was observed upon cysteine (Cys) addition while the emission band at 405 nm experienced a slight increase. The introduction of ACM into the probe led to the recovery of fluorescence at 630 nm, while the fluorescence at 405 nm remained largely unaffected. This selective response provides a ratiometric method for determining ACM. A linear relationship between the response ratio (F630/F405) and ACM concentration was established in the range of 0.12–450 µM, with LOD of 0.03 µM. The fluorescent probe boasts several advantages, including a low LOD, robustness against interference, and reliability. Furthermore, the synthesized probe was deployed for detecting ACM in certain food matrices. The recovery percentages ranged from 96.5 to 102.6 %, with RSD between 2.98 and 4.87 %, thus affirming the potential practical applications of the fluorescent probe.

Amperometric Detection of Acrylamide in Fried Food using Green-Synthesized Silver Nanoparticles

Acrylamide, a potential toxin and possible carcinogen, is formed in starchy foods cooked at high temperatures and poses a serious food safety concern. This study presents an amperometric sensor using green-synthesized silver nanoparticles (AgNPs) from neem (Azadirachta indica) leaves for the detection of acrylamide (Acr) in fried foods, with a focus on fried plantain chips. The use of AgNPs in conjunction with the specificity of hemoglobin (Hb) provides a rapid and accurate detection mechanism for Acr. Characterization of the AgNPs revealed their crystalline nature and excellent electron transfer properties, which are essential for effective sensor functionality. The sensor responded quickly and was able to accurately identify acrylamide concentrations as low as 5 mM within just 10 s. In addition, it demonstrated high precision, stability, and robust correlation with high-performance liquid chromatography analyzes in real food samples, confirming its practicality as an essential tool for ensuring food safety and quality control. This study highlights the benefits of green synthesis in acrylamide sensing and promotes the use of environmentally friendly materials in food safety applications.

Acrylamide detection and reduction in meat products using organic acids, fruit extracts, and probiotics

Grilled foods are an important source of acrylamide, which has neurotoxic, genotoxic, and carcinogenic properties. The current study aims to evaluate the level of acrylamide in beef, chicken, and fish products, especially those requiring high cooking temperatures, using High Performance Liquid Chromatography (HPLC). Reduction of acrylamide by organic acids i.e., (citric acid, malic acid, tartaric acid, and lactic acid) and fruit extracts of lemon, apple, and grape has also been investigated. The results revealed that the highest mean acrylamide concentration was found in chicken products (grilled chicken) which recorded 8.32 μg/100 g, followed by beef products (beef grilled) with a concentration of 7.91 μg/100 g, and fish products (pan-fried fish burgers) which recorded 6.77 μg/100 g). Furthermore, the mixture of organic acid has the highest effect on reducing the level of acrylamide in a chemical model system. Moreover, the fruit extract mixture was more effective in reducing the percentage of acrylamide in the grilled chicken than organic acids mixture. Finally, the addition of fruit extract improved the sensory properties of grilled chickens. In sum, this study offers novel and promising natural strategies to decrease acrylamide in meat products toward further future application in meat industry to deliver safe food to consumers.

Graphene quantum dots mediated acrylamide biosensing

Acrylamide (AA) is an unwanted food processing contaminant produced during cooking at high temperatures through the Maillard reaction between asparagine and reducing sugars or reactive carbonyls. In humans, it causes significant neurotoxicity, genotoxicity, and possible carcinogenicity. AA is categorized as a ‘Group 2A’ carcinogen by the International Agency for Research on Cancer (IARC). Bread, biscuits, and potato crisps are examples of foods heavy in starch that contain significant quantities of AA. Consequently, the detrimental effects of AA on human health make determining its quantitative content a crucial matter. Here, by immobilizing Hemoglobin nanoparticles (HbNPs) onto graphene quantum dots (GQDs) decorated pencil graphite (PG) electrode, we have developed a highly sensitive electrochemical biosensor for the detection of AA. Using cyclic voltammetry (CV), scanning electron microscopy (SEM), and electrochemical impedance spectroscopy (EIS), the developed working electrode (HbNPs/GQDs/PG) was examined at different stages of its preparation. The ideal pH and temperature for the fabricated biosensor were 5.0 and 35 °C, respectively and the biosensor demonstrated an acceptable level of selectivity, sensitivity, and stability along with a broad linear range (10 nM to 120 nM) and a low detection limit of 2.70 nM. The suggested strategy made use of less costly raw materials to ensure that it should be both effective and environmentally friendly. The sensor was effectively used to measure the amounts of AA in potato crisp samples.

Advancements in Chemical and Biosensors for Point-of-Care Detection of Acrylamide.

Acrylamide (AA), an odorless and colorless organic small-molecule compound found generally in thermally processed foods, possesses potential carcinogenic, neurotoxic, reproductive, and developmental toxicity. Compared with conventional methods for AA detection, bio/chemical sensors have attracted much interest in recent years owing to their reliability, sensitivity, selectivity, convenience, and low cost. This paper provides a comprehensive review of bio/chemical sensors utilized for the detection of AA over the past decade. Specifically, the content is concluded and systematically organized from the perspective of the sensing mechanism, state of selectivity, linear range, detection limits, and robustness. Subsequently, an analysis of the strengths and limitations of diverse analytical technologies ensues, contributing to a thorough discussion about the potential developments in point-of-care (POC) for AA detection in thermally processed foods at the conclusion of this review.

Construction and characterization of murine single-chain variable fragment (MuscFv) antibody against acrylamide in coffee

Ingredients of food, especially sugar and starch at high-temperature cooking processes could lead to the formation of acrylamide (AA). This chemical is a harmful carcinogen, a neurotoxicant, a reproductive toxicant, and a carcinogen in animal species. However, the detection of acrylamide contamination in food goes unnoticed. In this work, the mouse monoclonal antibody in the form of a single chain variable fragment (scFv) specific to acrylamide was selected from the murine scfv (muscfv) phage-displayed library. Acrylamide (AA) was used as an antigen for bio-panning. The murine single-chain variable fragment (MuscFv) antibody specific to acrylamide in coffee was successfully constructed, which was determined by ELISA and HPLC. Currently, this is the first study, which describes the selection of antibodies against acrylamide from the muscfv phage-displayed library and could be used as a tool for the detection of acrylamide in coffee.

Dimensionality engineering of flower-like bimetallic nanozyme with high peroxidase-activity for naked-eye and on-site detection of acrylamide in thermally processed foods

Acrylamide (AA) is a neurotoxin and carcinogen that formed during the thermal food processing. Conventional quantification techniques are difficult to realize on-site detection of AA. Herein, a flower-like bimetallic FeCu nanozyme (FeCuzyme) sensor and portable platform were developed for naked-eye and on-site detection of AA. The FeCuzyme was successfully prepared and exhibited flower-like structure with 3D catalytic centers. Fe/Cu atoms were considered as active center and ligand frameworks were used as cofactor, resulting in collaborative substrate-binding features and remarkably peroxidase-like activity. During the catalytic process, the 3,3′,5,5′-tetrame-thylbenzidine (TMB) oxidation can be quenched by glutathione (GSH), and then restored after thiolene Michael addition reaction between GSH and AA. Given the “on–off–on” effect for TMB oxidation and high POD-like activity, FeCuzyme sensor exhibited a wide linear relationship from 0.50 to 18.00 ​μM ​(R2 ​= ​0.9987) and high sensitivity (LOD ​= ​0.2360 ​μM) with high stability. The practical application of FeCuzyme sensor was successfully validated by HPLC method. Furthermore, a FeCuzyme portable platform was designed with smartphone/laptop, and which can be used for naked-eye and on-site quantitative determination of AA in real food samples. This research provides a way for rational design of a novel nanozyme-based sensing platform for AA detection.

Derivation of a no significant risk level (NSRL) of acrylamide in potato-based synthetic models and validation by NIR spectroscopy

The composition of raw potato-based products, mainly sugars and amino acids, plays a vital role in acrylamide formation after thermal treatment. The aim of the present study was to derive a No Significant Risk Level (NSRL) of acrylamide in potato-based synthetic models and validate it using Near Infrared Radiation spectroscopy (NIR). Therefore, NIR spectroscopy was chosen to validate gas chromatography Flame-Ionization Detector (GC-FID) concerning acrylamide detection in synthetic potato models and real potatoes. Subsequently, the Megazyme kit, High-Performance Liquid Chromatography, and GC-FID were applied to measure sugars, amino acids, and acrylamide. Findings revealed that peaks at 4439-4477 cm-1 were assigned to indicate acrylamide presence in all potato samples. Moreover, sucrose and asparagine had a significant impact on acrylamide. The prototype model unveiled the best combination among all potato models regarding the least acrylamide formation (Agria > Monalisa > Kennebec > Prototype). Mathematical equations were derived to calculate acrylamide concentration that should be below the threshold LOQ < 166.6 µg.kg-1, thus indicating an NSRL of acrylamide in the produced potato snacks if Agria and Monalisa cultivars were used.

SERS Sensor Based on Core–Shell Au@Ag Nanoparticles for the Sensitive Detection of Acrylamide in Foods

SERS Sensor Based on Core–Shell Au@Ag Nanoparticles for the Sensitive Detection of Acrylamide in Foods

Dietary Acrylamide: A Detailed Review on Formation, Detection, Mitigation, and Its Health Impacts.

In today's fast-paced world, people increasingly rely on a variety of processed foods due to their busy lifestyles. The enhanced flavors, vibrant colors, and ease of accessibility at reasonable prices have made ready-to-eat foods the easiest and simplest choice to satiate hunger, especially those that undergo thermal processing. However, these foods often contain an unsaturated amide called 'Acrylamide', known by its chemical name 2-propenamide, which is a contaminant formed when a carbohydrate- or protein-rich food product is thermally processed at more than 120 °C through methods like frying, baking, or roasting. Consuming foods with elevated levels of acrylamide can induce harmful toxicity such as neurotoxicity, hepatoxicity, cardiovascular toxicity, reproductive toxicity, and prenatal and postnatal toxicity. This review delves into the major pathways and factors influencing acrylamide formation in food, discusses its adverse effects on human health, and explores recent techniques for the detection and mitigation of acrylamide in food. This review could be of interest to a wide audience in the food industry that manufactures processed foods. A multi-faceted strategy is necessary to identify and resolve the factors responsible for the browning of food, ensure safety standards, and preserve essential food quality traits.

Determination of Acrylamide in Coffee, Cereals, Baby Food, Cocoa, Dry Pet Food, Potato Products, Vegetable Crisps, Biscuits, Tea, Nuts, and Spices by LC-MS/MS in a Single-Laboratory Validation: First Action 2023.01.

Acrylamide (AA) is a process contaminant naturally formed during the cooking of starchy food at high temperatures. Considering existing risks of misquantification inherent to the analysis of AA, an AOAC initiative raised the need for a consensus standard to determine AA in a broad variety of food. A quantitative LC-MS/MS method for AA determination in food was validated in a single-laboratory study. Targeted performance requirements in terms of target matrixes, limit of quantification, recovery, and precision were as defined per Standard Method Performance Requirement (SMPR®) 2022.006. The proposed method derives from EN 16618:2015 standard pending modifications brought to the (1) sample preparation (simplified, potentially automated); (2) scope of application (significantly extended); and (3) LC conditions (improved selectivity). Confirmatory detection of AA is conducted by LC-MS/MS in the Selected Reaction Monitoring mode (SRM), and isotopic dilution was applied for quantification approach using either 2,3,3-d3-acrylamide (d3-AA), or 13C3-2,3,3-d3-acrylamide (13C3-d3-AA) as labeled internal standard. A total of 16 laboratory samples from nine matrix categories were included in the validation process. A full validation was conducted on coffee (instant, roast), infant cereal, cocoa powder, pet food (croquettes), tea (green tea), spices (black pepper), and nuts (roasted almonds) with satisfactory performances both in terms of recovery (97-108%) and precision (RSDr and RSDiR <12%). The method applicability was further demonstrated through the analysis of quality control materials and reference materials including French fries, potato crisps, vegetable crisps, instant coffee, infant food, and biscuits (cookies), with accuracy values determined within a 94-107% range. The performances of the presented method are in agreement with the acceptance criteria stipulated in SMPR 2022.006. The Expert Review Panel for acrylamide approved the present method as AOAC Official First Action 2023.01.

Acrylamide in food products and the role of electrochemical biosensors in its detection: a comprehensive review.

In this review, the mechanisms of acrylamide formation in food, along with aspects related to its toxicity and associated consumption risks, are investigated, highlighting the potential impact on human health. European regulations regarding acrylamide content in food products are also addressed, emphasizing the importance of monitoring and detecting this substance in nutrition, by public health protection measures. The primary objective of the research is to explore and analyze innovative methods for detecting acrylamide in food, with a particular focus on electrochemical biosensors. This research direction is motivated by the need to develop rapid, sensitive, and efficient monitoring techniques for this toxic compound in food products, considering the associated consumption risks. The research has revealed several significant results. Studies have shown that electrochemical biosensors based on hemoglobin exhibited increased sensitivity and low detection limits, capable of detecting very low concentrations of acrylamide in processed foods. Additionally, it has been found that the use of functionalized nanomaterials, such as carbon nanotubes and gold nanoparticles, has led to the improvement of electrochemical biosensor performance in acrylamide detection. The integration of these technological innovations and functionalization strategies has enhanced the sensitivity, specificity, and stability of biosensors in measuring acrylamides. Thus, the results of this research offer promising perspectives for the development of precise and efficient methods for monitoring acrylamides in food, contributing to the improvement of food quality control and the protection of consumer health.

Advancements in Fluorescence Sensing: Carbon Quantum Dots for Acrylamide Detection in Food

Acrylamide is a hazardous chemical mainly synthesized during the thermal processing of foods representing a significant concern within the broader issue of food contaminants and their impact on public health. Acrylamide can be absorbed by the human body through dietary intake, respiration, dermal contact, and mucosa. The metabolic conversion of acrylamide into mercapturic acid metabolites and glycidamide results in several adverse and toxic effects. Therefore, this review explores the formation, toxicity, and metabolism of acrylamide. Hence, it is crucial to detect and ensure product quality via risk evaluation. Traditional analytical techniques for acrylamide detection often require expensive instrumentation and complex sample preparation, prompting the exploration of alternative, cost‐effective, sustainable methods. Here, we propose the utilization of carbon quantum dots (CQDs) synthesized through green approaches as a novel solution. CQDs display their immense potential for diverse applications due to their valuable properties such as biocompatibility, photocatalysis, and strong fluorescence. This review highlights the distinct potential of CQDs as a fluorescence probe for detecting acrylamide, showcasing their efficacy in addressing food safety concerns. In addition, various extraction and purification techniques for acrylamide such as QuEChERS, solid phase extraction, Carrez clarification, and dispersive liquid‐liquid microextraction are comprehensively reviewed. QuEChERS is regarded as a most promising technique for the extraction of acrylamide owing to its cost‐effective, rapid, and higher recovery rates.

Enhanced detection of acrylamide using a versatile solid-state upconversion sensor through spectral and visual analysis

Acrylamide (AM) generally forms in high-temperature processes and has been classified as a potential carcinogen. In this study, we put forward a maneuverable solid-state luminescence sensor using polydimethylsiloxane (PDMS) as the matrix coupled with upconversion nanoparticles as the indicator. The core-shell upconversion nanoparticles emitting cyan light were uniformly encapsulated in PDMS. Then it was further modified with complementary DNA of AM aptamer. The nanocrystalline fluorescein isothiocyanate isomer (FITC), coupled with AM aptamer, was attached to the surface of PDMS. FITC effectively quenched the upconversion luminescence through fluorescence resonance energy transfer (FRET). The introduction of AM resulted in preferentially bound to aptamer caused the separation of the quencher and the donor, and led to luminescence recovery. The developed sensor was applied for both spectral and visual monitoring, demonstrating a detection limit (LOD) of 1.00 nM and 1.07 nM, respectively. Importantly, in the actual foodstuffs detection, there is no obvious difference between the results of this study and the standard method, which indicates the developed method has good accuracy. Therefore, this solid-state sensor has the potential for on-site detection using a smartphone device and an Android application.

Optimization, Validation, and Application of a Quantitative GC/NPD Method for Acrylamide Determination in Yemeni Fried Fish Samples

The matrix's complexity makes measuring acrylamide in food items challenging, requiring a time-consuming extraction technique and high method sensitivity. A homemade solid phase extraction (SPE) device and a gas chromatography/nitrogen phosphorus detector (GC/NPD) were used to establish a simple and inexpensive acrylamide detection method in fried fish. Optimization and validation were used to measure acrylamide concentrations in twelve fried fish samples, four of which were home-fried and eight from Sana'a, Yemen, restaurants. The validated technique has high linearity R2 (0.995) in the 0.02–4 ppm range of acrylamide standards and good sensitivity with LOD (0.0062 ppm), substantially below the authorized level. The twelve samples had acrylamide values of 0.0583–0.4643 ppm. Due to variations in thermal treatments during frying, market-fried fish samples had greater acrylamide concentrations than home-fried fish samples. In the current study, the improved simple extraction process and GC/NPD protocol were useful for acrylamide measurement and may be employed as a prescreening tool for food sector acrylamide quantification.

Detection of acrylamide in food based on MIL-glucose oxidase cascade colorimetric aptasensor

BackgroundMaillard reaction involves the polymerization, condensation, and other reactions between compounds containing free amino groups and reducing sugars or carbonyl compounds during heat processing. This process endows unique flavors and colors to food, while it can also produce numerous hazards. Acrylamide (AAm) is one of Maillard's hazards with neurotoxicity and carcinogenicity, these effects can trigger mutations and alternations in gene expression in human cells and accelerate organ aging. An accurate and reliable acrylamide detection method with high sensitivity and specificity for future regulatory activities is urgently needed. ResultsHerein, we constructed a colorimetric aptasensor with the hybridization of MIL-glucose oxidase (MGzyme)-cDNA and magnetic nanoparticle-aptamer (MNP-Apt) to specifically detect AAm. The incorporation of MB-Apt and AAm released MGzyme-cDNA in the supernatant, took the supernatant out, with the addition of glucose and TMB, MGzyme would oxidize glucose, the resulting •OH facilitated the oxidation of colorless TMB to blue ox-TMB. The absorbance value at 652 nm, which indicates the characteristic absorption peak of ox-TMB, exhibited a proportion to the concentration of AAm. MGzyme avoided the addition of harmful intermediate H2O2 and created an acid microenvironment for the catalytic reaction. MNP-Apt possessed the advantages of high specificity and simplified separation. Under optimal conditions, this method displayed a linear range of 0.01–100 μM with the limit of detection of 1.53 nM. With the spiked analysis data cross-verified by ELISA kit, this aptasensor was proven to specifically detect AAm at low concentrations. SignificanceThis colorimetric aptasensor was the integration of aptamer and the enzyme-cascade system, which could broaden the applicable range of enzyme-cascade system, break the limits of specific detection of substrates, eliminate the need for harmful intermediates, improve the reaction efficiency, implement the specific detection, whilst enabling the accurate detection of AAm. Given these remarkable performances, this method has shown significant potential in the field of food safety inspection.

Polypyrrole participates in the construction of a polarity-switchable photoelectrochemical molecularly imprinted sensor for the detection of acrylamide in fried foods

Designing a polarity-switchable photoelectrochemical (PEC) sensor is important, but there are significant challenges due to the difficulty in adjusting the polarization of photoelectric or photoactive materials. In this manuscript, we established a simple and polarity-switchable molecularly imprinted polymer (MIP)-PEC sensor based on a “Z-scheme” strategy for detecting acrylamide (AM). First, we prepared a composite material of MoS2/rGO/Au (MGA) with a large specific surface area and good electrical conductivity that was sensitive to visible light. Second, the photoactive material MGA was used as the substrate material and polypyrrole (PPy) was acted as the functional monomer associated with the template molecule AM to construct the MIP working electrodes. PPy not only dressed up as a functional monomer but also as a polarity switcher, creating an in-situ-grown PPy/MGA heterojunction with a charge-carrier transport path similar to the letter “Z.” The design of the Z-scheme strategy not only increased the signal intensity of the photocurrent but also switched the photocurrent polarity from the anode to the cathode, avoiding the interference of a false positive or a false negative. When using this PEC-MIP sensor for the quantitative detection of AM, the detection limit was 1.7 pmol for the linear range of 0.25–10000 nmol. The proposed sensor exhibited satisfactory sensitivity and selectivity, and it has great potential for monitoring emerging food safety risks.

Detection and Quantification of Acrylamide in Second Trimester Amniotic Fluid Using a Novel LC-MS/MS Technique to Determine Whether High Acrylamide Content during Pregnancy Is Associated with Fetal Growth.

Acrylamide, an organic compound, is, chemically speaking, a vinyl-substituted primary amide. It is produced industrially, principally as a precursor to polyacrylamides, for use in such products as plastics and cosmetics. This same compound, however, forms naturally in certain foods, both home-cooked and packaged, especially when prepared at high temperatures. We developed and validated a novel reliable technique for the determination of acrylamide in amniotic fluid. Multiple reaction monitoring (MRM) is a targeted mass spectrometry (MS) technique which enables the detection and quantification of particular molecules in a complex mixture. Thanks to its throughput, selectivity, and sensitivity, MRM-MS has been identified as offering an alternative to antibody-based studies for the purpose of biomarker verification. Our aim was to investigate the presence of acrylamide in amniotic fluid and, via the MRM-MS technique, to determine whether there is any correlation between maternal exposure to acrylamide, through a woman's diet, and fetal growth. Our amniotic fluid bank included 40 samples from various fetal growth rates, as objectively denoted by the neonatal weight centile at delivery, while our analytical detection method was based on liquid chromatography-tandem mass spectrometry (LC-MS/MS). Acrylamide was determined with reversed phase chromatography and monitoring of two multiple reaction monitoring (MRM) transitions. Quantification was performed using the matrix-matched calibration curve. Acrylamide was detected at concentrations between 7.1 and 1468 ng/mL in six out of the total of 40 amniotic fluid samples that were used. Our method limit of detection and quantification was 1.4 ng/mL and 4.6 ng/mL, respectively. The repeatability of our method ranged between 11 and 14%, expressed as relative standard deviation levels between 5 and 100 ng/mL. Detection of acrylamide in early second trimester amniotic fluid, for the first time in the literature to our knowledge, raises concerns about fetal health, given that published data on animal studies have attributed a number of birth defects to acrylamide. Our novel LC-MS/MS method for the determination of acrylamide in amniotic fluid proved to be effective and its performance in practice was very accurate, simple, and fast. Validation of the method revealed that the use of a matrix-matched curve is necessary for the quantification.