{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T08:43:05Z","timestamp":1793090585790,"version":"4.5.0"},"reference-count":27,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2023,11,13]],"date-time":"2023-11-13T00:00:00Z","timestamp":1699833600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>Introduction: 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\u2019s diet, and fetal growth. Methods: 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. Results: 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. Conclusions: 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.<\/jats:p>","DOI":"10.3390\/biology12111425","type":"journal-article","created":{"date-parts":[[2023,11,13]],"date-time":"2023-11-13T08:35:52Z","timestamp":1699864552000},"page":"1425","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["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"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9067-5308","authenticated-orcid":false,"given":"Nikolaos","family":"Vrachnis","sequence":"first","affiliation":[{"name":"Third Department of Obstetrics and Gynecology, General University Hospital \u201cAttikon\u201d, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nikolaos","family":"Loukas","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Tzaneio General Hospital, 18536 Piraeus, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1333-4267","authenticated-orcid":false,"given":"Nikolaos","family":"Antonakopoulos","sequence":"additional","affiliation":[{"name":"Third Department of Obstetrics and Gynecology, General University Hospital \u201cAttikon\u201d, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece"},{"name":"Department of Obstetrics and Gynecology, University Hospital of Patras, Medical School, University of Patras, 26500 Patra, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2464-5108","authenticated-orcid":false,"given":"Niki","family":"Maragou","sequence":"additional","affiliation":[{"name":"Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zographou, 15771 Athens, Greece"},{"name":"Department of Food Science and Technology, University of West Attica, 12243 Egaleo, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7775-4634","authenticated-orcid":false,"given":"Marios","family":"Kostakis","sequence":"additional","affiliation":[{"name":"Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zographou, 15771 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-3278-7461","authenticated-orcid":false,"given":"Aliki","family":"Tsakni","sequence":"additional","affiliation":[{"name":"Department of Food Science and Technology, University of West Attica, 12243 Egaleo, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dionysios","family":"Vrachnis","sequence":"additional","affiliation":[{"name":"Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Despina","family":"Vougiouklaki","sequence":"additional","affiliation":[{"name":"Department of Food Science and Technology, University of West Attica, 12243 Egaleo, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nikolaos","family":"Machairiotis","sequence":"additional","affiliation":[{"name":"Third Department of Obstetrics and Gynecology, General University Hospital \u201cAttikon\u201d, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arhodoula","family":"Chatzilazarou","sequence":"additional","affiliation":[{"name":"Department of Wine, Vine and Beverage Sciences, University of West Attica, 12243 Egaleo, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dimitra","family":"Houhoula","sequence":"additional","affiliation":[{"name":"Department of Food Science and Technology, University of West Attica, 12243 Egaleo, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9972-8901","authenticated-orcid":false,"given":"Rozeta","family":"Sokou","sequence":"additional","affiliation":[{"name":"Neonatal Intensive Care Unit, \u201cAgios Panteleimon\u201d General Hospital of Nikea, 18454 Nikea, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sofoklis","family":"Stavros","sequence":"additional","affiliation":[{"name":"Third Department of Obstetrics and Gynecology, General University Hospital \u201cAttikon\u201d, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Drakakis","sequence":"additional","affiliation":[{"name":"Third Department of Obstetrics and Gynecology, General University Hospital \u201cAttikon\u201d, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George","family":"Mastorakos","sequence":"additional","affiliation":[{"name":"Unit of Endocrinology, Diabetes Mellitus and Metabolism, Aretaieio Hospital, Medical School, National and Kapodistrian University of Athens, 11528 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4316-7305","authenticated-orcid":false,"given":"Zoi","family":"Iliodromiti","sequence":"additional","affiliation":[{"name":"Neonatal Department, Aretaieio Hospital, Medical School, National and Kapodistrian University of Athens, 11526 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,13]]},"reference":[{"key":"ref_1","unstructured":"(2022). Does Burnt Food Give You Cancer?, University of Birmingham. Available online: https:\/\/www.birmingham.ac.uk\/research\/perspective\/does-burnt-food-give-you-cancer.aspx."},{"key":"ref_2","unstructured":"(2021). Can Burnt Toast Cause Cancer?, Healthline. Available online: https:\/\/www.healthline.com\/nutrition\/burnt-toast-cancer."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"109771","DOI":"10.1016\/j.foodcont.2023.109771","article-title":"Evaluation of the effects of intermittent frying in French fries and frying oil on monochloropropanediols, glycidols and acrylamide","volume":"150","author":"Pantalone","year":"2023","journal-title":"Food Control"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Pogurschi, E.N., Zugravu, C.A., Ranga, I.N., Trifunschi, S., Munteanu, M.F., Popa, D.C., Tudorache, M., and Custura, I. (2021). Determination of Acrylamide in Selected Foods from the Romanian Market. Foods, 10.","DOI":"10.3390\/foods10092110"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"121933","DOI":"10.1016\/j.jchromb.2019.121933","article-title":"Determination of acrylamide in gingerbread and other food samples by HILIC-MS\/MS: A dilute-and-shoot method","volume":"1136","author":"Sharma","year":"2020","journal-title":"J. Chromatogr. B"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"463651","DOI":"10.1016\/j.chroma.2022.463651","article-title":"Trace determination of prohibited acrylamide in cosmetic products by vortex-assisted reversed-phase dispersive liquid-liquid microextraction and liquid chromatography-tandem mass spectrometry","volume":"1687","author":"Schettino","year":"2023","journal-title":"J. Chromatogr. A"},{"key":"ref_7","unstructured":"(2019). Acrylamide and Cancer Risk, American Cancer Society. Available online: https:\/\/www.cancer.org\/cancer\/risk-prevention\/chemicals\/acrylamide.html."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2176","DOI":"10.1111\/1541-4337.12696","article-title":"Biomedical rationale for acrylamide regulation and methods of detection","volume":"20","author":"Cantrell","year":"2021","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_9","unstructured":"(2021). Can Eating Burnt Foods Cause Cancer?, Cancer Research UK. Available online: https:\/\/www.cancerresearchuk.org\/about-cancer\/causes-of-cancer\/cancer-myths\/can-eating-burnt-foods-cause-cancer."},{"key":"ref_10","unstructured":"(2015). Food Analysis\u2014Determination of Acrylamide in Food by Liquid Chromatography Tandem Mass Spectrometry (LC-ESI-MS\/MS) (Standard No. BS EN 16618:2015). Available online: https:\/\/standards.iteh.ai\/catalog\/standards\/cen\/cf79df49-9418-4fbe-a129-dfae52a4d9e4\/en-16618-2015."},{"key":"ref_11","unstructured":"Granby, K. (2019). Guidance Document on Analyses of Acrylamide in Food Including Analyses in the Low Concentration Range, Technical University of Denmark."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"460566","DOI":"10.1016\/j.chroma.2019.460566","article-title":"Sources of overestimation in the analysis of acrylamide-in coffee by liquid chromatography mass spectrometry","volume":"1610","author":"Desmarchelier","year":"2020","journal-title":"J. Chromatogr. A"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Vrachnis, N., Loukas, N., Vrachnis, D., Antonakopoulos, N., Zygouris, D., K\u03bflialexi, A., Pergaliotis, V., Iavazzo, C., Mastrakos, G., and Iliodromiti, Z. (2021). A Systematic Review of Bisphenol A from Dietary and Non-Dietary Sources during Preg-nancy and Its Possible Connection with Fetal Growth Restriction: Investigating Its Potential Effects and the Window of Fetal Vulnerability. Nutrients, 13.","DOI":"10.3390\/nu13072426"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4985","DOI":"10.1080\/14767058.2021.1873943","article-title":"Phthalates and fetal growth velocity: Tracking down the suspected links","volume":"35","author":"Vrachnis","year":"2021","journal-title":"J. Matern. Neonatal. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"6887","DOI":"10.18632\/aging.204269","article-title":"Fetal programming: In utero exposure to acrylamide leads to intergenerational disrupted ovarian function and accelerated ovarian aging","volume":"14","author":"Aldawood","year":"2022","journal-title":"Aging"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1002\/uog.19073","article-title":"Fetal Medicine Foundation fetal and neonatal population weight charts","volume":"52","author":"Nicolaides","year":"2018","journal-title":"Ultrasound Obstet. Gynecol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"8476217","DOI":"10.1155\/2018\/8476217","article-title":"Association between Brain-Derived Neu-rotrophic Factor (BDNF) Levels in 2nd Trimester Amniotic Fluid and Fetal Development","volume":"2018","author":"Antonakopoulos","year":"2018","journal-title":"Mediat. Inflamm."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Vrachnis, D., Antonakopoulos, N., Fotiou, A., Pergialiotis, V., Loukas, N., Valsamakis, G., Iavazzo, C., Stavros, S., Maroudias, G., and Panagopoulos, P. (2023). Is There a Correlation between Apelin and Insulin Concentrations in Early Second Trimester Amniotic Fluid with Fetal Growth Disorders?. J. Clin. Med., 12.","DOI":"10.3390\/jcm12093166"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1791","DOI":"10.1017\/S1368980019005123","article-title":"Relationship between gestational acrylamide exposure and offspring\u2019s growth: A systematic review and meta-analysis of cohort studies","volume":"23","author":"Zhan","year":"2020","journal-title":"Public Health Nutr."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1038\/sj.jp.7211290","article-title":"Amniotic Fluid: Not Just Fetal Urine Anymore","volume":"25","author":"Underwood","year":"2005","journal-title":"J. Perinatol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0890-6238(02)00078-3","article-title":"Effects of acrylamide on rodent reproductive performance","volume":"17","author":"Tyl","year":"2003","journal-title":"Reprod. Toxicol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1002\/bdrb.20030","article-title":"NTP-CERHR expert panel report on the reproductive and developmental toxicity of acrylamide","volume":"74","author":"Manson","year":"2005","journal-title":"Birth Defects Res. Part B: Dev. Reprod. Toxicol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.nut.2010.11.005","article-title":"Effects of fried potato chip supplementation on mouse pregnancy and fetal development","volume":"27","author":"Gaur","year":"2011","journal-title":"Nutrition"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.mrgentox.2004.09.008","article-title":"The carcinogenicity of acrylamide","volume":"580","author":"Rice","year":"2005","journal-title":"Mutat. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1739","DOI":"10.1289\/ehp.1205327","article-title":"Birth weight, head circumference, and prenatal exposure to acrylamide from maternal diet: The European prospective mother-child study (NewGeneris)","volume":"120","author":"Pedersen","year":"2012","journal-title":"Environ. Health Perspect."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1486","DOI":"10.1542\/peds.2005-2629","article-title":"The influence of head growth in fetal life, infancy, and childhood on intelligence at the ages of 4 and 8 years","volume":"118","author":"Gale","year":"2008","journal-title":"Pediatrics"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1186\/s12940-021-00715-0","article-title":"Cord blood acrylamide levels and birth size, and interactions with genetic variants in acrylamide-metabolising genes","volume":"20","author":"Hogervorst","year":"2021","journal-title":"Environ. Health"}],"container-title":["Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-7737\/12\/11\/1425\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:22:08Z","timestamp":1760131328000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-7737\/12\/11\/1425"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,13]]},"references-count":27,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2023,11]]}},"alternative-id":["biology12111425"],"URL":"https:\/\/doi.org\/10.3390\/biology12111425","relation":{},"ISSN":["2079-7737"],"issn-type":[{"value":"2079-7737","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,11,13]]}}}