Department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100 Sıhhıye, Ankara, Turkey.
Department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, 06100 Sıhhıye, Ankara, Turkey.
J Pharm Biomed Anal. 2020 Sep 5;188:113427. doi: 10.1016/j.jpba.2020.113427. Epub 2020 Jul 2.
Down Syndrome is a genetic disorder caused by the presence of all or part of a third copy of chromosome 21. Metabolomics is identification and quantification of small-molecule metabolites (molecular weight <1000 Da) in tissues, cells and physiological fluids within a certain period time. Metabolites are intermediate products of various types of biochemical reactions that participate in bonding metabolic pathways. In this study, metabolites such as 2-Hydroxybutyric acid, 3-Hydroxybutyric acid, β-Hydroxyisovaleric acid, Uracil, Glutamic acid, Maltose and Melezitose were chosen as the possible determinants/markers for the prenatal screening of Down Syndrome. Quantitative analysis of the metabolites conducted by GCMS method using 5 % phenyl / 95 % dimethylpolysiloxane (30 m ×0.25 mm, 0.25 μm film thickness) capillary column. The oven temperature was held constant at 60 °C for 1 min and ramped at 10 °C /min to 200 °C then ramped at 30 °C/min to 320 °C and hold for 6 min before cool-down, as helium mobile phase and flow rate of 2.8 mL/min and adding Myristic acid-d27 as an internal standard. Our method was validated by parameters of system suitability, stability, linearity, sensitivity, accuracy, precision, selectivity, robustness and ruggedness. The developed and validated method was applied to plasma samples taken from pregnant women with Down Syndrome (study group) and euploid fetuses (healthy group). The levels of these seven metabolites are statistically different (p < 0.05 for all) between the groups. It can be concluded that these relevant metabolites might be used for the prenatal screening of Down Syndrome.
唐氏综合征是一种由 21 号染色体全部或部分三体引起的遗传疾病。代谢组学是在一定时间内鉴定和定量组织、细胞和生理流体中小分子代谢物（分子量<1000 Da）的方法。代谢物是参与连接代谢途径的各种类型生化反应的中间产物。在这项研究中，选择 2-羟基丁酸、3-羟基丁酸、β-羟基异戊酸、尿嘧啶、谷氨酸、麦芽糖和密二糖等代谢物作为唐氏综合征产前筛查的可能决定因素/标志物。使用 5%苯基/95%二甲基聚硅氧烷（30 m×0.25 mm，0.25 μm 薄膜厚度）毛细管柱，通过 GCMS 方法对代谢物进行定量分析。在 60°C 下保持恒温 1 分钟，然后以 10°C/min 的速度升温至 200°C，然后以 30°C/min 的速度升温至 320°C，并在冷却前保持 6 分钟，氦作为流动相，流速为 2.8 mL/min，并添加肉豆蔻酸-d27 作为内标。我们的方法通过系统适用性、稳定性、线性、灵敏度、准确性、精密度、选择性、稳健性和耐用性的参数进行验证。开发和验证的方法应用于唐氏综合征孕妇（研究组）和正常胎儿（健康组）的血浆样本。两组间这七种代谢物的水平存在统计学差异（p<0.05）。可以得出结论，这些相关代谢物可能用于唐氏综合征的产前筛查。
