Heralde Francisco M, Martin Zaidy T, Cagayan Maria Stephanie Fay S, Uy Esterlita V, Ubial Paulyn Jean R, Velarde Michael C, Llamas-Clark Erlidia F
Department of Biochemistry and Molecular Biology, College of Medicine, University of the Philippines Manila.
Department of Pharmacology and Toxicology, College of Medicine, University of the Philippines Manila.
Acta Med Philipp. 2025 Feb 28;59(3):70-79. doi: 10.47895/amp.vi0.9007. eCollection 2025.
Endocrine Disrupting Chemicals (EDCs) are ubiquitously found as low-level contaminants and pose serious threat to women's health. EDCs may result in various reproductive disorders, fetal birth and developmental abnormalities, and endocrine and metabolic disorders. EDCs can be detected in body fluids of exposed individuals including blood and urine. This study aimed to detect four EDCs - Methyl Paraben (MP), 2,4-Dichlorophenoxyacetic acid (2,4-D), Monobutyl Phthalate (MBP), and Bisphenol A (BPA) in urine samples of women using Ultra-Performance Liquid Chromatography - Quadrupole Time-of-Flight (UPLC-QTOF) mass spectrometry.
Sequential steps of enzymatic deconjugation, liquid-liquid extraction, solid phase extraction, and liquid chromatography separation and mass spectrometry detection were optimized in urine samples. The method was used to analyze 70 urine samples from women of reproductive age.
The sample preparation method showed a recovery ranging from 86.6% (MBP) to 100 % (2,4-D). The method demonstrated limits of quantitation ranging from 1.52 ng/m(MP) to 6.46 ng/mL(2,4D). Intra-day precisions expressed as relative standard deviation were all below 15% while accuracy was shown to range from 67.10% (2,4-D) to 102.39% (MBP). MP was detected in nine samples (12.86%) with a geometric mean value of 10.15 ng/ml (range: 3.62-52.39 ng/ml). MBP was detected in 68 samples (97.14%) with a geometric mean value of 97.62 ng/ml (range: 15.32-698.18 ng/ml). BPA was detected only once (9.58 ng/ml) while 2, 4-D was not detected in all samples.
A UPLC-QTOF mass spectrometry method to detect four EDCs at parts per billion level (ng/ml) was adapted and applied for analysis of urine samples. This method can find applicability in routine testing of clinical specimens as well as surveillance and other epidemiological studies.
内分泌干扰物（EDCs）作为低水平污染物广泛存在，对女性健康构成严重威胁。EDCs可能导致各种生殖障碍、胎儿出生及发育异常，以及内分泌和代谢紊乱。在包括血液和尿液在内的暴露个体的体液中可检测到EDCs。本研究旨在使用超高效液相色谱-四极杆飞行时间（UPLC-QTOF）质谱法检测女性尿液样本中的四种EDCs——对羟基苯甲酸甲酯（MP）、2,4-二氯苯氧乙酸（2,4-D）、邻苯二甲酸单丁酯（MBP）和双酚A（BPA）。
对尿液样本中酶解共轭、液液萃取、固相萃取以及液相色谱分离和质谱检测的连续步骤进行了优化。该方法用于分析70例育龄女性的尿液样本。
样品制备方法的回收率在86.6%（MBP）至100%（2,4-D）之间。该方法的定量限在1.52 ng/m（MP）至6.46 ng/mL（2,4-D）之间。以相对标准偏差表示的日内精密度均低于15%，而准确度在67.10%（2,4-D）至102.39%（MBP）之间。在9个样本（12.86%）中检测到MP，几何平均值为10.15 ng/ml（范围：3.62 - 52.39 ng/ml）。在68个样本（97.14%）中检测到MBP，几何平均值为97.62 ng/ml（范围：15.32 - 698.18 ng/ml）。仅一次检测到BPA（9.58 ng/ml），而所有样本中均未检测到2,4-D。
采用UPLC-QTOF质谱法在十亿分之一水平（ng/ml）检测四种EDCs，并将其应用于尿液样本分析。该方法可应用于临床标本的常规检测以及监测和其他流行病学研究。
