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Simultaneous determination of 10 phthalate metabolites in urine by ultra performance liquid chromatography -tandem mass spectrometry
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Xiaoying ZHAO1, Hui KANG2, Yanyan WANG1, Wenhui LI3, Ming CHEN3, Chan KE1, Feng QIN1, *, Xixiong KANG2, *
Chinese Journal of Chromatography | 2025, 43(6) : 640 - 649
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Chinese Journal of Chromatography | 2025, 43(6): 640-649
• Articles •
Simultaneous determination of 10 phthalate metabolites in urine by ultra performance liquid chromatography -tandem mass spectrometry
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Xiaoying ZHAO1, Hui KANG2, Yanyan WANG1, Wenhui LI3, Ming CHEN3, Chan KE1, Feng QIN1, *, Xixiong KANG2, *
Affiliations
  • 1. Analytical Chemistry Laboratory,Shenyang Pharmaceutical University,Shenyang 110000,China
  • 2. Beijing Zhongjian IVD Engineering Research Center,Beijing 100095,China
  • 3. Yujing Technology Shanghai Co. ,Ltd. ,Shanghai 200120,China
Published: 2025-06-08 doi: 10.3724/SP.J.1123.2024.04002
Outline
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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.

ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)  /  phthalate metabolites  /  urine samples  /  plasticizer
赵小英, 康慧, 王燕燕, 李文慧, 陈铭, 可婵, 秦峰, 康熙雄. 超高效液相色谱-串联质谱同时测定尿液中10种邻苯二甲酸酯代谢物. 色谱, 2025 , 43 (6) : 640 -649 . DOI: 10.3724/SP.J.1123.2024.04002
Xiaoying ZHAO, Hui KANG, Yanyan WANG, Wenhui LI, Ming CHEN, Chan KE, Feng QIN, Xixiong KANG. Simultaneous determination of 10 phthalate metabolites in urine by ultra performance liquid chromatography -tandem mass spectrometry[J]. Chinese Journal of Chromatography, 2025 , 43 (6) : 640 -649 . DOI: 10.3724/SP.J.1123.2024.04002
  • National Key Research and Development Program of China(2022YFC2009600)
  • National Key Research and Development Program of China(2022YFC2009603)
Year 2025 volume 43 Issue 6
30
16
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Article Info
doi: 10.3724/SP.J.1123.2024.04002
  • Receive Date:2024-04-01
  • Online Date:2026-10-30
  • Published:2025-06-08
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  • Received:2024-04-01
Funding
National Key Research and Development Program of China(2022YFC2009600)
National Key Research and Development Program of China(2022YFC2009603)
Affiliations
    1. Analytical Chemistry Laboratory,Shenyang Pharmaceutical University,Shenyang 110000,China
    2. Beijing Zhongjian IVD Engineering Research Center,Beijing 100095,China
    3. Yujing Technology Shanghai Co. ,Ltd. ,Shanghai 200120,China
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表12种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
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属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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