Department of Physical Chemistry, Medical University of Lublin, Chodźki 4a, 20-093 Lublin, Poland.
Doctoral School of Medical University of Lublin, Medical University of Lublin, Chodźki 7, 20-093 Lublin, Poland.
Int J Environ Res Public Health. 2022 Feb 17;19(4):2309. doi: 10.3390/ijerph19042309.
Amniocentesis involves taking a sample of the amniotic fluid in order to perform a karyotype test and diagnose any genetic defects that may affect the fetus. Amniotic fluid has been collected from patients with an indication for amniocentesis in the 15-26th week of pregnancy. A simple and sensitive high-performance liquid chromatography with fluorescence detection (HPLC-FLD) method for identification and quantification of eleven selected bisphenols in amniotic fluid samples is proposed. The proposed method involved protein precipitation using acetonitrile, and next the extraction and concentration of analytes by solid-phase extraction (SPE). The solid-phase extraction (SPE) procedure with application of Oasis HLB SPE columns performed well for the majority of the analytes, with recoveries in the range of 67-121% and relative standard deviations (RSD%) less than 16%. The limits of detection (LODs) and quantification (LOQs) of all the investigated analytes were in the range of 0.8-2.5 ng mL and 2.4-7.5 ng mL (curves constructed in methanol) and 1.1-5.2 ng mL and 3.2-15.6 ng mL (curves constructed in the amniotic fluid), respectively. The method was validated at the following two concentration levels: 10 ng mL (2 × LOQ) and 20 ng mL (4 LOQ). The results confirm the validity of the SPE procedure and HPLC-FLD method for identification and quantification of bisphenols in amniotic fluid samples collected during an amniocentesis. The result obtained show that HPLC-FLD is a useful method for determination of bisphenol residues at nanogram per milliliter concentrations in amniotic fluid samples. Residues of five analytes (BADGE·2HO, BPAF, BADGE, BADGE·HO·HCl and BADGE·2HCl) were detected in amniotic fluid samples. Additionally, the harmfulness of bisphenols as potential pathogens that may cause karyotype disorders and contribute to preterm birth was estimated.
羊膜穿刺术涉及抽取羊水样本，以进行核型测试并诊断可能影响胎儿的任何遗传缺陷。在妊娠 15-26 周时，对有羊膜穿刺术指征的患者采集羊水。提出了一种简单灵敏的高效液相色谱-荧光检测法（HPLC-FLD），用于鉴定和定量羊水样品中的 11 种选定的双酚。该方法涉及使用乙腈进行蛋白质沉淀，然后通过固相萃取（SPE）提取和浓缩分析物。应用 Oasis HLB SPE 柱的固相萃取（SPE）程序对大多数分析物表现良好，回收率在 67-121%范围内，相对标准偏差（RSD%）小于 16%。所有研究分析物的检测限（LOD）和定量限（LOQ）均在 0.8-2.5 ng mL 和 2.4-7.5 ng mL（甲醇中构建的曲线）和 1.1-5.2 ng mL 和 3.2-15.6 ng mL（羊水构建的曲线）范围内。该方法在以下两个浓度水平进行了验证：10 ng mL（2×LOQ）和 20 ng mL（4 LOQ）。结果证实了 SPE 程序和 HPLC-FLD 方法在鉴定和定量羊水中双酚的有效性收集在羊膜穿刺术中。结果表明，HPLC-FLD 是一种有用的方法，可用于测定羊水样品中双酚残留物的纳克/毫升浓度。在羊水样品中检测到五种分析物（BADGE·2HO、BPAF、BADGE、BADGE·HO·HCl 和 BADGE·2HCl）的残留。此外，还评估了双酚作为可能导致染色体异常和导致早产的潜在病原体的危害性。
