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Simultaneous identification of three emergent stimulant laxative adulterants in slimming foods using only one antibody

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Abstract

Stimulant laxatives were recently found to be abused in slimming foods, resulting in harmful effects on consumers. To ensure the safety of relative products, sensitive yet multiplex immunoassays are crucial in rapid screening of stimulant laxatives. However, there are few immunoassays for these substances, and even less for broad-specific recognition. Thus, in this work, four theoretically promising haptens of emerging stimulant laxative bisacodyl were rationally designed using molecular modeling and synthesized to immune animals, whose feasibility was confirmed by the obtained broad-specific antibody. Based on this unique antibody, a highly sensitive multiplex competitive indirect enzyme-linked immunosorbent assay (ciELISA) was established with low limits of detection for bisacodyl, sodium picosulfate, and BHPM (0.23, 13.68, and 0.11 ng/mL). In spiked sample recovery test and real sample detection, this ciELISA exhibited acceptable consistency with the validation method, demonstrating high accuracy and applicability of our method. This reliable multiplex ciELISA proceeds the rapid screening of stimulant laxatives in slimming foods.

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Abbreviations

BHPM:

bis-(p-Hydroxyphenyl)-pyridyl-2-methane

BSA:

Bovine serum albumin

CDI:

NN′-Carbonyldiimidazole

CR:

Cross-reactivity

ciELISA:

Competitive indirect enzyme-linked immunosorbent assay

DMF:

N,N-Dimethylformamide

EDC:

1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride

GC-MS:

Gas chromatography-mass spectrometry

HPLC:

High-performance liquid chromatographic

KLH:

Keyhole limpet hemocyanin

LC-MS/MS:

Liquid chromatography–tandem mass spectrometry

LF:

Lactoferrin

NHS:

N-Hydroxysuccinimide

NMR:

Nuclear magnetic resonance

OVA:

Ovalbumin

TMB:

3,3′,5,5′-Tetramethylbenzidine

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Funding

This work was financially supported by National Key Research and Development Program of China (2023YFF1104700, 2023YFF1105100), National Scientific Foundation of China (31871883), Guangdong Marine Economic Development Program for Six Categories of Marine Industries (no. GDNRC[2023]36), Guangdong Natural Science Foundation (2023A1515011834), Generic Technique Innovation Team Construction of Modern Agriculture of Guangdong Province (2023KJ130), Zhongshan Science and Technology Innovation Team (no. CXTD2022004), Yangjiang Science and Technology Program (SDZX20220003), and Project from Guangdong Laboratory for Lingnan Modern Agriculture (nos. LNSYSZX001 and P20211154-0102-02).

Author information

Author notes

  1. Tian Guan and Sha Chen contributed equally to this work.

Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Food Quality and Safety/National-Local Joint Engineering Research Center for Machining and Safety of Livestock and Poultry Products, South China Agricultural University, 483 Wushan Road, Tianhe District, Guangzhou, 510642, China

    Tian Guan, Sha Chen & Hongtao Lei

  2. Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou, 510642, China

    Tian Guan & Hongtao Lei

  3. Guangzhou Institute of Food Inspection, Guangzhou, 510080, China

    Yanhong Chen & Yu Wang

  4. Sichuan Institute of Food Inspection, Chengdu, 610097, China

    Xiaoqin Yu

  5. Tropical Medicine Institute and South China Chinese Medicine Collaborative Innovation Center, Guangzhou University of Chinese Medicine, Guangzhou, 510405, China

    Xinan Huang

  6. Guangzhou Customs Technology Center Food and Cosmetics Testing Institute, Guangzhou, 510075, China

    Qing Liu

  7. Guangzhou Huibiao Testing Technology Center, 412 Jinlingnan Road, Nansha District, Guangzhou, 511458, China

    Qunhua Yang

Authors
  1. Tian Guan
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Contributions

Tian Guan: investigation, methodology, validation, writing — original draft.

Sha Chen: conceptualization, data curation, formal analysis, investigation, methodology, validation, writing — original draft.

Yanhong Chen: validation.

Yu Wang: validation.

Xiaoqin Yu: methodology.

Xinan Huang: investigation, methodology.

Qing Liu: validation.

Hongtao Lei: conceptualization, funding acquisition, supervision, writing — review and editing.

Qunhua Yang: funding acquisition, supervision, writing – review and editing.

Corresponding authors

Correspondence to Hongtao Lei or Qunhua Yang.

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Ethics approval

All animals were treated according to regulations approved by the Laboratory Animal Care of South China Agricultural University. The ethical review of animal experiments (Animal Experiment Ethical Approval Number: 2020e009) can be found in Fig. S1.

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The authors declare no competing interests.

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Guan, T., Chen, S., Chen, Y. et al. Simultaneous identification of three emergent stimulant laxative adulterants in slimming foods using only one antibody. Anal Bioanal Chem 416, 2995–3006 (2024). https://doi.org/10.1007/s00216-024-05246-4

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