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
Nikolaos Vrachnis,
Nikolaos Loukas,
Nikolaos Antonakopoulos,
Niki Maragou,
Marios Kostakis,
Aliki Tsakni,
Dionysios Vrachnis,
Despina Vougiouklaki,
Nikolaos Machairiotis,
Arhodoula Chatzilazarou,
Dimitra Houhoula,
Rozeta Sokou,
Sofoklis Stavros,
Peter Drakakis,
George Mastorakos,
Zoi Iliodromiti
Affiliations
Nikolaos Vrachnis
Third Department of Obstetrics and Gynecology, General University Hospital “Attikon”, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece
Nikolaos Loukas
Department of Obstetrics and Gynecology, Tzaneio General Hospital, 18536 Piraeus, Greece
Nikolaos Antonakopoulos
Third Department of Obstetrics and Gynecology, General University Hospital “Attikon”, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece
Niki Maragou
Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zographou, 15771 Athens, Greece
Marios Kostakis
Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zographou, 15771 Athens, Greece
Aliki Tsakni
Department of Food Science and Technology, University of West Attica, 12243 Egaleo, Greece
Dionysios Vrachnis
Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece
Despina Vougiouklaki
Department of Food Science and Technology, University of West Attica, 12243 Egaleo, Greece
Nikolaos Machairiotis
Third Department of Obstetrics and Gynecology, General University Hospital “Attikon”, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece
Arhodoula Chatzilazarou
Department of Wine, Vine and Beverage Sciences, University of West Attica, 12243 Egaleo, Greece
Dimitra Houhoula
Department of Food Science and Technology, University of West Attica, 12243 Egaleo, Greece
Rozeta Sokou
Neonatal Intensive Care Unit, “Agios Panteleimon” General Hospital of Nikea, 18454 Nikea, Greece
Sofoklis Stavros
Third Department of Obstetrics and Gynecology, General University Hospital “Attikon”, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece
Peter Drakakis
Third Department of Obstetrics and Gynecology, General University Hospital “Attikon”, Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece
George Mastorakos
Unit of Endocrinology, Diabetes Mellitus and Metabolism, Aretaieio Hospital, Medical School, National and Kapodistrian University of Athens, 11528 Athens, Greece
Zoi Iliodromiti
Neonatal Department, Aretaieio Hospital, Medical School, National and Kapodistrian University of Athens, 11526 Athens, Greece
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’s 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.
