Skip to main content
SpringerLink
Log in

Investigation of diethylstilbestrol residue level in human urine samples by a specific monoclonal antibody

  • Research Article
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Diethylstilbestrol (DES) is used as a kind of animal feed additive and affects people’s health through the food chain. The purpose of this study is to detect the residue level of DES in 576 human urine samples directly. DES-BSA was used to immunize Balb/c mice. The monoclonal antibody was produced by hybridoma that was screened through cell fusion techniques. Finally, we developed the indirect competitive ELISA method to analyze 576 human urine samples from Zhejiang Province, China. The IC50 of this method was 3.33 ng/mL. The LOD and LOQ were 0.16 and 0.54 ng/mL. Linear range of the standard curve was from LOD to 12.50 ng/mL. There was no cross-reactivity with two kinds of estrogens and two structural analogs with DES. Five hundred seventy-six urine samples were analyzed by the indirect competitive ELISA method, and the detection rate was 98.78%. The mean concentration and geometric mean were 4.70 and 3.50 ng/mL. The indirect competitive ELISA method based on monoclonal antibody was sensitive and reliable for the detection of DES in human urine samples. The results warned us to pay more attention to human health and food safety.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Almeida C, Nogueira JM (2006) Determination of steroid sex hormones in water and urine matrices by stir bar sorptive extraction and liquid chromatography with diode array detection. J Pharm Biomed Anal 41:1303–1311

    Article  CAS  Google Scholar 

  • Aman CS, Pastor A, Cighetti G, de la Guardia M (2006) Development of a multianalyte method for the determination of anabolic hormones in bovine urine by isotope-dilution GC-MS/MS. Anal Bioanal Chem 386:1869–1879

    Article  CAS  Google Scholar 

  • Cooper J, Currie W, Elliott CT (2001) Comparison of the efficiences of enzymatic and chemical hydrolysis of (nortestosterone and diethylstilboestrol) glucuronides in bovine urine. J Chromatogr B Biomed Sci Appl 757:221–227

    Article  CAS  Google Scholar 

  • Dodds EC, Goldberg L, Lawson W, Robinson R (1938) Oestrogenic activity of certain synthetic compounds. Nature 141:247–248

    Article  CAS  Google Scholar 

  • Fernandez-Arauzo L, Pimentel-Trapero D, Hernandez-Carrasquilla M (2014) Simultaneous determination of resorcylic acid lactones, beta and alpha trenbolone and stilbenes in bovine urine by UHPLC/MS/MS. J Chromatogr B Analyt Technol Biomed Life Sci 973C:89–96

    Article  CAS  Google Scholar 

  • Ganan J, Morante-Zarcero S, Perez-Quintanilla D, Marina ML, Sierra I (2016) One-pot synthesized functionalized mesoporous silica as a reversed-phase sorbent for solid-phase extraction of endocrine disrupting compounds in milks. J Chromatogr A 1428:228–235

    Article  CAS  Google Scholar 

  • Gore AC, Chappell VA, Fenton SE, Flaws JA, Nadal A, Prins GS, Toppari J, Zoeller RT (2015) Executive Summary to EDC-2: The Endocrine Society’s Second Scientific Statement on Endocrine-Disrupting Chemicals. Endocr Rev 36:593–602

    Article  CAS  Google Scholar 

  • Han H, Kim B, Lee SG, Kim J (2013) An optimised method for the accurate determination of zeranol and diethylstilbestrol in animal tissues using isotope dilution-liquid chromatography/mass spectrometry. Food Chem 140:44–51

    Article  CAS  Google Scholar 

  • Hatch EE, Troisi R, Palmer JR, Wise LA, Titus L, Strohsnitter WC, Ricker W, Hyer M, Hoover RN (2015) Prenatal diethylstilbestrol exposure and risk of obesity in adult women. J Dev Orig Health Dis 6:201–207

    Article  CAS  Google Scholar 

  • Hatch EE, Herbst AL, Hoover RN, Noller KL, Adam E, Kaufman RH, Palmer JR, Titus-Ernstoff L, Hyer M, Hartge P, Robboy SJ (2001) Incidence of squamous neoplasia of the cervix and vagina in women exposed prenatally to diethylstilbestrol (United States). Cancer Causes Control 12:837–845

    Article  CAS  Google Scholar 

  • He X, Mei X, Wang J, Lian Z, Tan L, Wu W (2016) Determination of diethylstilbestrol in seawater by molecularly imprinted solid-phase extraction coupled with high-performance liquid chromatography. Mar Pollut Bull 102:142–147

    Article  CAS  Google Scholar 

  • Huo T, Wang L, Liu L, Chu X, Xu C (2006) Rapid time-resolved fluoroimmunoassay for diethylstilbestrol residues in chicken liver. Anal Biochem 357:272–276

    Article  CAS  Google Scholar 

  • Iparraguirre A, Navarro P, Rodil R, Prieto A, Olivares M, Etxebarria N, Zuloaga O (2014) Matrix effect during the membrane-assisted solvent extraction coupled to liquid chromatography tandem mass spectrometry for the determination of a variety of endocrine disrupting compounds in wastewater. J Chromatogr A 1356:163–170

    Article  CAS  Google Scholar 

  • Jiang X, Chen HQ, Cui ZH, Yin L, Zhang WL, Liu WB, Han F, Ao L, Cao J, Liu JY (2016) Low-dose and combined effects of oral exposure to bisphenol A and diethylstilbestrol on the male reproductive system in adult Sprague-Dawley rats. Environ Toxicol Pharmacol 43:94–102

    Article  CAS  Google Scholar 

  • Kumirska J, Plenis A, Lukaszewicz P, Caban M, Migowska N, Bialk-Bielinska A, Czerwicka M, Stepnowski P (2013) Chemometric optimization of derivatization reactions prior to gas chromatography-mass spectrometry analysis. J Chromatogr A 1296:164–178

    Article  CAS  Google Scholar 

  • Laronda MM, Unno K, Butler LM, Kurita T (2012) The development of cervical and vaginal adenosis as a result of diethylstilbestrol exposure in utero. Differentiation 84:252–260

    Article  CAS  Google Scholar 

  • Leena H-C (2014) Maternal exposure to diethylstilbestrol during pregnancy and increased breast cancer risk in daughters. Breast Cancer Res 16:208–217

    Google Scholar 

  • Lei Y, Zhang Q, Fang L, Akash MS, Rehman K, Liu Z, Shi W, Chen S (2014) Development and comparison of two competitive ELISAs for estimation of cotinine in human exposed to environmental tobacco smoke. Drug testing and analysis 6:1020–1027

    Article  CAS  Google Scholar 

  • Lv X, Xiao S, Zhang G, Jiang P, Tang F (2016) Occurrence and removal of phenolic endocrine disrupting chemicals in the water treatment processes. Sci Rep 6:1–10

    Article  Google Scholar 

  • Tedeschi CA, Rubin M, Krumholz BA (2005) Six cases of women with diethylstilbestrol in utero demonstrating long-term manifestations and current evaluation guidelines. J Low Genit Tract Dis 9:11–18

    Article  Google Scholar 

  • Wang X, Reisberg S, Serradji N, Anquetin G, Pham MC, Wu W, Dong CZ, Piro B (2014) E-assay concept: detection of bisphenol A with a label-free electrochemical competitive immunoassay. Biosens Bioelectron 53:214–219

    Article  CAS  Google Scholar 

  • Wu X, Wang L, Ma W, Zhu Y, Xu L, Kuang H, Xu C (2012) A simple, sensitive, rapid and specific detection method for bisphenol A based on fluorescence polarization immunoassay. Immunol Investig 41:38–50

    Article  CAS  Google Scholar 

  • Yang Q, Diamond MP, Al-Hendy A (2016) Early life adverse environmental exposures increase the risk of uterine fibroid development: role of epigenetic regulation. Front Pharmacol 7:40

    Google Scholar 

  • Yin L, Zheng LJ, Jiang X, Liu WB, Han F, Cao J, Liu JY (2015) Effects of low-dose diethylstilbestrol exposure on DNA methylation in mouse spermatocytes. PLoS One 10:1–18

    Google Scholar 

  • Zhang S, Lei Y, Xu X, Shi W, Chen S (2013) Synthesis of artificial diethylstilbestrol antigen for preparation of polyclonal antibodies. Zhejiang Da Xue Xue Bao Yi Xue Ban 42:25–31

    CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the State Key Program of National Natural Science of China (Grant No. 81430081), the grant (No. 2012ZX09506001-004) from the National Major Projects for Science and Technology Development of Ministry Science and Technology of China, and the Fundamental Research Funds for the Central Universities.

Author information

Authors and Affiliations

  1. Institute of Drug Metabolism and Pharmaceutical Analysis, College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Zijingang Campus, Hangzhou, Zhejiang, 310058, China

    Dai Zhang, Zhan Zhou, Jie Zhou & Shuqing Chen

  2. Hangzhou EPIE Bio-detection Technology Limited, Hangzhou, 310051, China

    Lifang Zhou & Yajing Lei

Authors
  1. Dai Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lifang Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yajing Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhan Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jie Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shuqing Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Jie Zhou or Shuqing Chen.

Ethics declarations

All experiments on mice were performed according to the ethics committee of the Experimental Animal Center of Zhejiang University.

Additional information

Responsible editor: Philippe Garrigues

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, D., Zhou, L., Lei, Y. et al. Investigation of diethylstilbestrol residue level in human urine samples by a specific monoclonal antibody. Environ Sci Pollut Res 24, 7042–7050 (2017). https://doi.org/10.1007/s11356-017-8405-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-017-8405-y

Keywords

Search

Navigation