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Ultra-High-Performance Liquid Chromatography-Isotope Dilution Tandem Mass Spectrometry for the Determination of Phthalate Secondary Metabolites in Human Serum Based on Solid-Phase Extraction

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Background: A multitude of biomonitoring analytical methods are applied in urinary detection, but few are applicable to phthalate metabolites in human serum, and those that are invariably involve complex operation. Objective: To develop a novel method for the determination of phthalate monoesters in human serum using solid-phase extraction (SPE) and ultra-HPLC (UHPLC)-tandem mass spectrometry for multiple human serum sample determination. Method: Analytes in serum samples were extracted and purified with a novel SPE cartridge named Prime HLB and then directly injected and analyzed without eluent redissolution. After sample extraction and cleanup, 11 phthalate monoesters were separated on a C18 chromatographic column in UHPLC. Among them, mono-isobutyl phthalate and mono-n-butyl phthalate, a pair of isomers, were successfully separated. Results: Lower LODs of the 11 monoesters in 0.5 mL human serum were in the range of 0.03–3 ng/mL, and lower LOQs accordingly ranged from 0.1 to 10.0 ng/mL. Recoveries from spiked bovine serum samples were in the range of 95.3–109.9%, and RSDs were 8.0–12.0%. Intra- and inter-day recoveries were in the range of 94.1–115.7%, with RSDs <13.7%. Conclusions: The proposed method has been proved to be rapid, simple, accurate, and sensitive, so that it may potentially be used for high-throughput biological monitoring with high efficiency and precision. Highlights: Sample pretreatment does not require eluent drying and redissolution. Mass losses of analytes are avoided. Operational procedures are easy and rapid. The method has proved applicable for 11 phthalate metabolites in multiple human serum sample determinations.
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Document Type: Research Article

Affiliations: 1: China National Center for Food Safety Risk Assessment, Beijing, China 10021 2: North China Institute of Science and Technology, Hebei, China 3: Shimadzu (China) Co., Shanghai, China

Publication date: January 1, 2019

This article was made available online on June 28, 2018 as a Fast Track article with title: "Ultra-High-Performance Liquid Chromatography-Isotope Dilution Tandem Mass Spectrometry for the Determination of Phthalate Secondary Metabolites in Human Serum Based on Solid-Phase Extraction".

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  • The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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