Zhao Xiao-Ying, Kang Hui, Wang Yan-Yan, Li Wen-Hui, Chen Ming, Ke Chan, Qin Feng, Kang Xi-Xiong
Analytical Chemistry Laboratory,Shenyang Pharmaceutical University,Shenyang 110000,China.
Beijing Zhongjian IVD Engineering Research Center,Beijing 100095,China.
Se Pu. 2025 Jun;43(6):640-649. doi: 10.3724/SP.J.1123.2024.04002.
Phthalates (PAEs) are widely employed as plasticizers in plastic products that are used in industrial, agricultural, food, medical, and other fields. PAEs are relatively weakly bonded to plastic products through non-covalent interactions. Consequently, PAEs can easily leak from the product into the environment, which exposes the public to PAEs through food intake, skin absorption from personal care products, and by inhaling air. Related studies have shown that PAEs are endocrine-disrupting substances and that long-term exposure to PAEs may result in diseases of the nervous, reproductive, cardiovascular and immune systems. In addition, excessive exposure to PAEs may trigger inflammatory responses and induce tumors. Therefore, establishing a highly sensitive assay for determining PAE levels in the human body following exposure is an important objective. PAEs generally have half-lives of less than 24 h; they are rapidly metabolized through enzymatic hydrolysis after entering the human body and excreted through urine. Therefore, most studies have focused on PAE metabolites as target compounds; hence, human body exposure to PAEs can be assessed by analyzing the types and levels of these metabolites. Herein, we established a method for simultaneously determining ten phthalate (PAE) metabolites in human urine using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The ten PAE metabolites in urine were separated using an ACQUITY UPLC BEH Phenyl column (50 mm×2.1 mm, 1.7 μm). Gradient elution was performed using 0.1% formic acid aqueous solution and 0.1% formic acid in acetonitrile as the mobile phases, at a flow rate of 0.5 mL/min, a column temperature of 40 ℃, and a sample size of 20 μL. Data were acquired in negative-ion electrospray ionization (ESI) and multiple reaction monitoring (MRM) modes, and quantified using the isotope internal standard method. The method was found to be highly specific, with the ten PAE metabolites exhibiting good linearities in their linear ranges, with limits of detection (LODs) and quantification (LOQs) of 0.03-0.3 and 0.1-1 ng/mL, respectively. Under the four quality control (QC) levels, the intra-day and inter-day precisions of the ten PAE metabolites were all ≤8.3%, and the accuracy ranged from ‒10.5% to 7.3%. The method was used to assess the exposure levels of PAE metabolites in the urine samples of 60 volunteers, with 1‒6 kinds of PAE metabolites detected in the urine of each volunteer. This method is sensitive, accurate, simple, efficient, and suitable for the large-scale biological monitoring of PAE metabolites.
邻苯二甲酸酯(PAEs)作为增塑剂被广泛应用于工业、农业、食品、医疗等领域的塑料制品中。PAEs通过非共价相互作用与塑料制品结合相对较弱。因此,PAEs很容易从产品中泄漏到环境中,公众可通过摄入食物、皮肤吸收个人护理产品以及吸入空气接触到PAEs。相关研究表明,PAEs是内分泌干扰物质,长期接触PAEs可能导致神经、生殖、心血管和免疫系统疾病。此外,过度接触PAEs可能引发炎症反应并诱发肿瘤。因此,建立一种高灵敏度的检测方法来测定人体接触PAEs后的体内水平是一个重要目标。PAEs的半衰期一般小于24小时;它们进入人体后通过酶促水解迅速代谢,并通过尿液排出。因此,大多数研究都集中在PAE代谢物作为目标化合物;因此,通过分析这些代谢物的类型和水平可以评估人体对PAEs的接触情况。在此,我们建立了一种使用超高效液相色谱-串联质谱(UPLC-MS/MS)同时测定人尿中十种邻苯二甲酸酯(PAE)代谢物的方法。尿中的十种PAE代谢物使用ACQUITY UPLC BEH苯基柱(50 mm×2.1 mm,1.7 μm)进行分离。以0.1%甲酸水溶液和0.1%甲酸乙腈溶液为流动相进行梯度洗脱,流速为0.5 mL/min,柱温为40℃,进样量为20 μL。数据在负离子电喷雾电离(ESI)和多反应监测(MRM)模式下采集,并使用同位素内标法进行定量。该方法具有高特异性,十种PAE代谢物在其线性范围内表现出良好的线性关系,检测限(LODs)和定量限(LOQs)分别为0.03 - 0.3和0.1 - 1 ng/mL。在四个质量控制(QC)水平下,十种PAE代谢物的日内和日间精密度均≤8.3%,准确度范围为 - 10.5%至7.3%。该方法用于评估60名志愿者尿液样本中PAE代谢物的接触水平,每名志愿者的尿液中检测到1 - 6种PAE代谢物。该方法灵敏、准确、简便、高效,适用于PAE代谢物的大规模生物监测。