Vrachnis Nikolaos, Loukas Nikolaos, Antonakopoulos Nikolaos, Maragou Niki, Kostakis Marios, Tsakni Aliki, Vrachnis Dionysios, Vougiouklaki Despina, Machairiotis Nikolaos, Chatzilazarou Arhodoula, Houhoula Dimitra, Sokou Rozeta, Stavros Sofoklis, Drakakis Peter, Mastorakos George, Iliodromiti Zoi
Third Department of Obstetrics and Gynecology, General University Hospital "Attikon", Medical School, National and Kapodistrian University of Athens, 12462 Athens, Greece.
Department of Obstetrics and Gynecology, Tzaneio General Hospital, 18536 Piraeus, Greece.
Biology (Basel). 2023 Nov 13;12(11):1425. doi: 10.3390/biology12111425.
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.
丙烯酰胺是一种有机化合物，从化学角度讲，它是一种乙烯基取代的伯酰胺。它主要在工业上作为聚丙烯酰胺的前体生产，用于塑料和化妆品等产品。然而，这种化合物在某些家庭烹饪和包装食品中自然形成，尤其是在高温制备时。我们开发并验证了一种用于测定羊水丙烯酰胺的新型可靠技术。多反应监测（MRM）是一种靶向质谱（MS）技术，能够检测和定量复杂混合物中的特定分子。由于其通量、选择性和灵敏度，MRM-MS已被确定为可作为基于抗体研究的替代方法用于生物标志物验证。我们的目的是研究羊水中丙烯酰胺的存在情况，并通过MRM-MS技术确定孕妇通过饮食接触丙烯酰胺与胎儿生长之间是否存在任何相关性。
我们的羊水库包含40个来自不同胎儿生长速率的样本，分娩时新生儿体重百分位数客观地表明了这些生长速率，而我们的分析检测方法基于液相色谱 - 串联质谱（LC-MS/MS）。采用反相色谱法和监测两个多反应监测（MRM）跃迁来测定丙烯酰胺。使用基质匹配校准曲线进行定量。
在所使用的40份羊水样本中，有6份检测到丙烯酰胺浓度在7.1至1468 ng/mL之间。我们方法的检测限和定量限分别为1.4 ng/mL和4.6 ng/mL。我们方法的重复性在11%至14%之间，以5至100 ng/mL之间的相对标准偏差水平表示。
据我们所知，首次在文献中报道在妊娠中期早期羊水中检测到丙烯酰胺，鉴于已发表的动物研究数据将多种出生缺陷归因于丙烯酰胺，这引发了对胎儿健康的担忧。我们用于测定羊水中丙烯酰胺的新型LC-MS/MS方法被证明是有效的，其在实际应用中的性能非常准确、简单且快速。该方法的验证表明，定量时使用基质匹配曲线是必要的。
