Development of a one-step centrifugal extraction method for simultaneous determination of multiple types of pesticides and their metabolites in urine, serum, and breast milk

•A one-step centrifugal-assisted CIPS method was developed in biological samples.•Centrifugal-assisted technique enhances performance in sample preparation.•Multiplexed tSIM method was used for highly sensitive and accurate quantification.•The greenness profile and practicality of this method were a...

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Published in:	Journal of Chromatography A Vol. 1763; p. 466432
Main Authors:	Zhang, Chi, Zhan, Zhipeng, Li, Jingguang, Wu, Yongning, Chen, Dawei, Li, Hong
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 22.11.2025
Subjects:	Advanced instrumental analysis Biomonitoring Centrifugation - methods Centrifuge Chromatography, Liquid - methods Female Humans Limit of Detection Linear Models Milk, Human - chemistry Multiplexed method Pesticide Residues - analysis Pesticide Residues - blood Pesticide Residues - isolation & purification Pesticide Residues - urine Pesticides - analysis Pesticides - blood Pesticides - isolation & purification Pesticides - metabolism Pesticides - urine Reproducibility of Results Sample preparation Centrifuge Biomonitoring Advanced instrumental analysis Sample preparation Multiplexed method
ISSN:	0021-9673, 1873-3778, 1873-3778
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Abstract	•A one-step centrifugal-assisted CIPS method was developed in biological samples.•Centrifugal-assisted technique enhances performance in sample preparation.•Multiplexed tSIM method was used for highly sensitive and accurate quantification.•The greenness profile and practicality of this method were assessed. An efficient and highly sensitive analytical method was developed for the simultaneous determination of neonicotinoids, benzopyrazoles, diamide insecticides and chlorothalonil fungicide and their metabolites in biological samples (urine, serum and breast milk). The method combines centrifugal-assisted cold-induced phase separation (CIPS) with liquid chromatography and high-resolution mass spectrometry (LCHRMS) in multiplexed-targeted single-ion monitoring (MSX-tSIM) mode to ensure high sensitivity and accuracy. Sample preparation parameters such as solvent selection and acetonitrile/water ratio were optimized to improve analyte recovery and minimize matrix effects. The method exhibited excellent accuracy, precision, and low limit of quantification (LLOQ) in the range of 0.01–0.2 µg/L, with recoveries ranging from 72.6 % to 109.6 % in a variety of biological matrices. The method showed good linearity (R² > 0.993) and was suitable for the detection of trace pesticide levels in complex biological samples. Biomonitoring of pooled samples revealed widespread neonicotinoids contamination, with fipronil metabolites detected only in serum and breast milk, raising concerns about the potential accumulation of pesticide metabolites in the human body, especially in susceptible populations. This method provides a cost-effective and environmentally friendly alternative to traditional methods, facilitates large-scale human biomonitoring studies, and supports pesticide exposure risk assessment.
AbstractList	An efficient and highly sensitive analytical method was developed for the simultaneous determination of neonicotinoids, benzopyrazoles, diamide insecticides and chlorothalonil fungicide and their metabolites in biological samples (urine, serum and breast milk). The method combines centrifugal-assisted cold-induced phase separation (CIPS) with liquid chromatography and high-resolution mass spectrometry (LCHRMS) in multiplexed-targeted single-ion monitoring (MSX-tSIM) mode to ensure high sensitivity and accuracy. Sample preparation parameters such as solvent selection and acetonitrile/water ratio were optimized to improve analyte recovery and minimize matrix effects. The method exhibited excellent accuracy, precision, and low limit of quantification (LLOQ) in the range of 0.01-0.2 µg/L, with recoveries ranging from 72.6 % to 109.6 % in a variety of biological matrices. The method showed good linearity (R² > 0.993) and was suitable for the detection of trace pesticide levels in complex biological samples. Biomonitoring of pooled samples revealed widespread neonicotinoids contamination, with fipronil metabolites detected only in serum and breast milk, raising concerns about the potential accumulation of pesticide metabolites in the human body, especially in susceptible populations. This method provides a cost-effective and environmentally friendly alternative to traditional methods, facilitates large-scale human biomonitoring studies, and supports pesticide exposure risk assessment.An efficient and highly sensitive analytical method was developed for the simultaneous determination of neonicotinoids, benzopyrazoles, diamide insecticides and chlorothalonil fungicide and their metabolites in biological samples (urine, serum and breast milk). The method combines centrifugal-assisted cold-induced phase separation (CIPS) with liquid chromatography and high-resolution mass spectrometry (LCHRMS) in multiplexed-targeted single-ion monitoring (MSX-tSIM) mode to ensure high sensitivity and accuracy. Sample preparation parameters such as solvent selection and acetonitrile/water ratio were optimized to improve analyte recovery and minimize matrix effects. The method exhibited excellent accuracy, precision, and low limit of quantification (LLOQ) in the range of 0.01-0.2 µg/L, with recoveries ranging from 72.6 % to 109.6 % in a variety of biological matrices. The method showed good linearity (R² > 0.993) and was suitable for the detection of trace pesticide levels in complex biological samples. Biomonitoring of pooled samples revealed widespread neonicotinoids contamination, with fipronil metabolites detected only in serum and breast milk, raising concerns about the potential accumulation of pesticide metabolites in the human body, especially in susceptible populations. This method provides a cost-effective and environmentally friendly alternative to traditional methods, facilitates large-scale human biomonitoring studies, and supports pesticide exposure risk assessment. •A one-step centrifugal-assisted CIPS method was developed in biological samples.•Centrifugal-assisted technique enhances performance in sample preparation.•Multiplexed tSIM method was used for highly sensitive and accurate quantification.•The greenness profile and practicality of this method were assessed. An efficient and highly sensitive analytical method was developed for the simultaneous determination of neonicotinoids, benzopyrazoles, diamide insecticides and chlorothalonil fungicide and their metabolites in biological samples (urine, serum and breast milk). The method combines centrifugal-assisted cold-induced phase separation (CIPS) with liquid chromatography and high-resolution mass spectrometry (LCHRMS) in multiplexed-targeted single-ion monitoring (MSX-tSIM) mode to ensure high sensitivity and accuracy. Sample preparation parameters such as solvent selection and acetonitrile/water ratio were optimized to improve analyte recovery and minimize matrix effects. The method exhibited excellent accuracy, precision, and low limit of quantification (LLOQ) in the range of 0.01–0.2 µg/L, with recoveries ranging from 72.6 % to 109.6 % in a variety of biological matrices. The method showed good linearity (R² > 0.993) and was suitable for the detection of trace pesticide levels in complex biological samples. Biomonitoring of pooled samples revealed widespread neonicotinoids contamination, with fipronil metabolites detected only in serum and breast milk, raising concerns about the potential accumulation of pesticide metabolites in the human body, especially in susceptible populations. This method provides a cost-effective and environmentally friendly alternative to traditional methods, facilitates large-scale human biomonitoring studies, and supports pesticide exposure risk assessment. An efficient and highly sensitive analytical method was developed for the simultaneous determination of neonicotinoids, benzopyrazoles, diamide insecticides and chlorothalonil fungicide and their metabolites in biological samples (urine, serum and breast milk). The method combines centrifugal-assisted cold-induced phase separation (CIPS) with liquid chromatography and high-resolution mass spectrometry (LCHRMS) in multiplexed-targeted single-ion monitoring (MSX-tSIM) mode to ensure high sensitivity and accuracy. Sample preparation parameters such as solvent selection and acetonitrile/water ratio were optimized to improve analyte recovery and minimize matrix effects. The method exhibited excellent accuracy, precision, and low limit of quantification (LLOQ) in the range of 0.01-0.2 µg/L, with recoveries ranging from 72.6 % to 109.6 % in a variety of biological matrices. The method showed good linearity (R² > 0.993) and was suitable for the detection of trace pesticide levels in complex biological samples. Biomonitoring of pooled samples revealed widespread neonicotinoids contamination, with fipronil metabolites detected only in serum and breast milk, raising concerns about the potential accumulation of pesticide metabolites in the human body, especially in susceptible populations. This method provides a cost-effective and environmentally friendly alternative to traditional methods, facilitates large-scale human biomonitoring studies, and supports pesticide exposure risk assessment.
ArticleNumber	466432
Author	Zhang, Chi Li, Hong Zhan, Zhipeng Li, Jingguang Wu, Yongning Chen, Dawei
Author_xml	– sequence: 1 givenname: Chi surname: Zhang fullname: Zhang, Chi organization: NHC Key Laboratory of Food Safety Risk Assessment, Chinese Academy of Medical Science Research Unit (No. 2019RU014), China National Center for Food Safety Risk Assessment, Beijing 100021, China – sequence: 2 givenname: Zhipeng surname: Zhan fullname: Zhan, Zhipeng email: zhanzhipeng@jzmu.edu.cn organization: School of Public Health, Jinzhou Medical University, Jinzhou 121001, China – sequence: 3 givenname: Jingguang surname: Li fullname: Li, Jingguang email: lijg@cfsa.net.cn organization: NHC Key Laboratory of Food Safety Risk Assessment, Chinese Academy of Medical Science Research Unit (No. 2019RU014), China National Center for Food Safety Risk Assessment, Beijing 100021, China – sequence: 4 givenname: Yongning surname: Wu fullname: Wu, Yongning email: wuyongning@cfsa.ent.cn organization: NHC Key Laboratory of Food Safety Risk Assessment, Chinese Academy of Medical Science Research Unit (No. 2019RU014), China National Center for Food Safety Risk Assessment, Beijing 100021, China – sequence: 5 givenname: Dawei orcidid: 0000-0002-4015-2557 surname: Chen fullname: Chen, Dawei email: chendw@cfsa.net.cn organization: NHC Key Laboratory of Food Safety Risk Assessment, Chinese Academy of Medical Science Research Unit (No. 2019RU014), China National Center for Food Safety Risk Assessment, Beijing 100021, China – sequence: 6 givenname: Hong surname: Li fullname: Li, Hong email: lih@jzmu.edu.cn organization: School of Public Health, Jinzhou Medical University, Jinzhou 121001, China
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Keywords	Centrifuge Biomonitoring Advanced instrumental analysis Sample preparation Multiplexed method
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Snippet	•A one-step centrifugal-assisted CIPS method was developed in biological samples.•Centrifugal-assisted technique enhances performance in sample... An efficient and highly sensitive analytical method was developed for the simultaneous determination of neonicotinoids, benzopyrazoles, diamide insecticides...
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SubjectTerms	Advanced instrumental analysis Biomonitoring Centrifugation - methods Centrifuge Chromatography, Liquid - methods Female Humans Limit of Detection Linear Models Milk, Human - chemistry Multiplexed method Pesticide Residues - analysis Pesticide Residues - blood Pesticide Residues - isolation & purification Pesticide Residues - urine Pesticides - analysis Pesticides - blood Pesticides - isolation & purification Pesticides - metabolism Pesticides - urine Reproducibility of Results Sample preparation
Title	Development of a one-step centrifugal extraction method for simultaneous determination of multiple types of pesticides and their metabolites in urine, serum, and breast milk
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