Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

A Stable Isotope Dilution Assay for Multi-class Antibiotics in Pregnant Urines by LC–MS/MS

  • 57 Accesses

Abstract

An accurate assay suitable for human biomonitoring of multiclass antibiotics preferred for veterinary use is currently lacking. In this study, we developed a liquid chromatography-triple quadrupole tandem mass spectrometry (LC–QqQ–MS/MS) method for analysis of 41 representative antibiotics (eight categories) and their two metabolites in human urine samples. Additionally, along with matrix-matched calibration and solid-phase extraction (SPE), use of 36 stable isotopically labeled internal standards (SIL-ISs) compensated for matrix effects and helped in the accuracy of quantitation. The absolute matrix effects (MEs), the linearity of limits of detection (LODs) and limits of quantitation (LOQs), stability, accuracy and precision of this method were validated. The validation parameters obtained for the targeted compounds are as follows: MEs (68.0–129.2%), LODs (0.016–1.481 ng mL−1), LOQs (0.050–4.812 ng mL−1), and higher recoveries (80.3–112.5%) except cyadox and cefaclor. The regression coefficients (r2) of targeted compounds at a concentration range of 0.05–100 ng mL−1 ranged from 0.9958 to 0.9998. The intra- and inter-precisions were obtained with a relative standard deviation (RSD%) < 13%. The method was successfully applied to analyze 302 urine samples of pregnant women from the China-Anhui Birth Cohort (C-ABC), and 36 of the selected antibiotics and two metabolites were detected. The total detection frequencies were 52.6% (fluoroquinolones), 40.4% (tetracyclines), 33.1% (sulfonamides), 16.9% (chloramphenicols), 11.6% (macrolides), 10.9% (β-lactams), 2.3% (lincosamides), and 0.3% (quinoxalines), respectively. The developed method has been successfully employed to obtain reliable quantitative results in the urine of pregnant women.

Graphic abstract

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

References

  1. 1.

    Qu X, Yin C, Sun X, Huang S, Li C, Dong P, Lu X, Zhang Z, Yin A (2018) Consumption of antibiotics in Chinese public general tertiary hospitals (2011–2014): trends, pattern changes and regional differences. PLoS ONE 13(5):e0196668. https://doi.org/10.1371/journal.pone.0196668

  2. 2.

    Yin J, Li Q, Sun Q (2018) Antibiotic consumption in Shandong Province, China: an analysis of provincial pharmaceutical centralized bidding procurement data at public healthcare institutions, 2012–2016. J Antimicrob Chemother 73(3):814–820. https://doi.org/10.1093/jac/dkx469

  3. 3.

    Qiao M, Ying GG, Singer AC, Zhu YG (2018) Review of antibiotic resistance in China and its environment. Environ Int 110:160–172. https://doi.org/10.1016/j.envint.2017.10.016

  4. 4.

    Wei R, He T, Zhang S, Zhu L, Shang B, Li Z, Wang R (2019) Occurrence of seventeen veterinary antibiotics and resistant bacterias in manure-fertilized vegetable farm soil in four provinces of China. Chemosphere 215:234–240. https://doi.org/10.1016/j.chemosphere.2018.09.152

  5. 5.

    Sandegren L (2019) Low sub-minimal inhibitory concentrations of antibiotics generate new types of resistance. Sustain Chem Pharm 11:46–48. https://doi.org/10.1016/j.scp.2018.12.006

  6. 6.

    Ben Y, Fu C, Hu M, Liu L, Wong MH, Zheng C (2019) Human health risk assessment of antibiotic resistance associated with antibiotic residues in the environment: a review. Environ Res 169:483–493. https://doi.org/10.1016/j.envres.2018.11.040

  7. 7.

    Zhou L, Limbu SM, Shen ML, Zhai WY, Qiao F, He AY, Du ZY, Zhang ML (2018) Environmental concentrations of antibiotics impair zebrafish gut health. Environ Pollut 235:245–254. https://doi.org/10.1016/j.envpol.2017.12.073

  8. 8.

    Founou LL, Founou RC, Essack SY (2016) Antibiotic resistance in the food chain: a developing country-perspective. Front Microbiol 7:1881. https://doi.org/10.3389/fmicb.2016.01881

  9. 9.

    Ortqvist AK, Lundholm C, Halfvarson J, Ludvigsson JF, Almqvist C (2019) Fetal and early life antibiotics exposure and very early onset inflammatory bowel disease: a population-based study. Gut 68(2):218–225. https://doi.org/10.1136/gutjnl-2017-314352

  10. 10.

    Muanda FT, Sheehy O, Berard A (2017) Use of antibiotics during pregnancy and the risk of major congenital malformations: a population based cohort study. Br J Clin Pharmacol 83(11):2557–2571. https://doi.org/10.1111/bcp.13364

  11. 11.

    Wang H, Yang J, Yu X, Zhao G, Zhao Q, Wang N, Jiang Y, Jiang F, He G, Chen Y, Zhou Z, Jiang Q (2018) Exposure of adults to antibiotics in a shanghai suburban area and health risk assessment: a biomonitoring-based study. Environ Sci Technol 52(23):13942–13950. https://doi.org/10.1021/acs.est.8b03979

  12. 12.

    Barr DB, Wang RY, Needham LL (2005) Biologic monitoring of exposure to environmental chemicals throughout the life stages: requirements and issues for consideration for the National Children's Study. Environ Health Perspect 113(8):1083–1091. https://doi.org/10.1289/ehp.7617

  13. 13.

    Panuwet P, Hunter RE Jr, D'Souza PE, Chen X, Radford SA, Cohen JR, Marder ME, Kartavenka K, Ryan PB, Barr DB (2016) Biological matrix effects in quantitative tandem mass spectrometry-based analytical methods: advancing biomonitoring. Crit Rev Anal Chem 46(2):93–105. https://doi.org/10.1080/10408347.2014.980775

  14. 14.

    May L, Chiu WL, Snyder ST, Wong PK, Liao JC, Gau V (2010) Matrix effects-a challenge toward automation of molecular analysis. J Assoc Lab Autom 15(3):233–242. https://doi.org/10.1016/j.jala.2010.02.001

  15. 15.

    Trufelli H, Palma P, Famiglini G, Cappiello A (2011) An overview of matrix effects in liquid chromatography–mass spectrometry. Mass Spectrom Rev 30(3):491–509. https://doi.org/10.1002/mas.20298

  16. 16.

    Jia S, Xu T, Huan T, Chong M, Liu M, Fang W, Fang M (2019) Chemical isotope labeling exposome (CIL-EXPOSOME): one high-throughput platform for human urinary global exposome characterization. Environ Sci Technol 53(9):5445–5453. https://doi.org/10.1021/acs.est.9b00285

  17. 17.

    Sharma P, Kumar D, Mutnuri S (2019) UPLC-MS/MS method validation of ciprofloxacin in human urine: application to biodegradability study in microbial fuel cell. Biomed Chromatogr 33(2):e4392. https://doi.org/10.1002/bmc.4392

  18. 18.

    Wang X, Guo T, Wei Y, Xu G, Li N, Feng J, Zhao R (2019) Determination of quinolone antibiotic residues in human serum and urine using high-performance liquid chromatography/tandem mass spectrometry. J Anal Toxicol. https://doi.org/10.1093/jat/bkz034

  19. 19.

    Chen C, Yan H, Shen BH, Zhuo XY (2011) Simultaneous determination of sixteen antibiotics in human urine with ultra performance liquid chromatography–tandem mass spectrometry. Fa Yi Xue Za Zhi 27(1):25–29

  20. 20.

    Wang HX, Wang B, Zhou Y, Jiang QW (2014) Rapid and sensitive screening and selective quantification of antibiotics in human urine by two-dimensional ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Anal Bioanal Chem 406(30):8049–8058. https://doi.org/10.1007/s00216-014-8197-6

  21. 21.

    Cazorla-Reyes R, Romero-Gonzalez R, Frenich AG, Rodriguez Maresca MA, Martinez Vidal JL (2014) Simultaneous analysis of antibiotics in biological samples by ultra high performance liquid chromatography–tandem mass spectrometry. J Pharm Biomed Anal 89:203–212. https://doi.org/10.1016/j.jpba.2013.11.004

  22. 22.

    Li N, Ho KWK, Ying GG, Deng WJ (2017) Veterinary antibiotics in food, drinking water, and the urine of preschool children in Hong Kong. Environ Int 108:246–252. https://doi.org/10.1016/j.envint.2017.08.014

  23. 23.

    Wang HX, Wang N, Qian JH, Hu LY, Huang PX, Su MF, Yu X, Fu CW, Jiang F, Qi Z, Ying Z, Lin HJ, He GS, Yue C, Jiang QW (2017) Urinary antibiotics of pregnant women in Eastern China and cumulative health risk assessment. Environ Sci Technol 51(6):3518–3525. https://doi.org/10.1021/acs.est.6b06474

  24. 24.

    Huang W, Qiu Q, Chen M, Shi J, Huang X, Kong Q, Long D, Chen Z, Yan S (2019) Determination of 18 antibiotics in urine using LC–QqQ–MS/MS. J Chromatogr B Analyt Technol Biomed Life Sci 1105:176–183. https://doi.org/10.1016/j.jchromb.2018.12.019

  25. 25.

    Liu S, Zhao G, Zhao H, Zhai G, Chen J, Zhao H (2017) Antibiotics in a general population: relations with gender, body mass index (BMI) and age and their human health risks. Sci Total Environ 599–600:298–304. https://doi.org/10.1016/j.scitotenv.2017.04.216

  26. 26.

    Liu HC, Lin T, Lin X, Shao JL, Li QW (2018) QuEChERS with magnetic hydrophilic-lipophilic balanced adsorbent and its application in multi-class veterinary residues in milk by ultra high-performance liquid chromatography–tandem mass spectrometry. Chromatographia 81(2):265–275. https://doi.org/10.1007/s10337-017-3433-9

  27. 27.

    Wang H, Wang N, Wang B, Zhao Q, Fang H, Fu C, Tang C, Jiang F, Zhou Y, Chen Y, Jiang Q (2016) Antibiotics in drinking water in shanghai and their contribution to antibiotic exposure of school children. Environ Sci Technol 50(5):2692–2699. https://doi.org/10.1021/acs.est.5b05749

  28. 28.

    Zhao Y, Zhou Y, Zhu Q, Xia B, Ma W, Xiao X, Shi H, Zhang Y (2019) Determination of antibiotic concentration in meconium and its association with fetal growth and development. Environ Int 123:70–78. https://doi.org/10.1016/j.envint.2018.11.053

  29. 29.

    Chiesa LM, Nobile M, Panseri S, Arioli F (2017) Antibiotic use in heavy pigs: comparison between urine and muscle samples from food chain animals analysed by HPLC–MS/MS. Food Chem 235:111–118. https://doi.org/10.1016/j.foodchem.2017.04.184

  30. 30.

    Zhu YD, Gao H, Huang K, Zhang YW, Cai XX, Yao HY, Mao LJ, Ge X, Zhou SS, Xu YY, Jin ZX, Sheng J, Yan SQ, Pan WJ, Hao JH, Zhu P, Tao FB (2018) Prenatal phthalate exposure and placental size and shape at birth: a birth cohort study. Environ Res 160:239–246. https://doi.org/10.1016/j.envres.2017.09.012

  31. 31.

    Tao FB, Hao JH, Huang K, Su PY, Cheng DJ, Xing XY, Huang ZH, Zhang JL, Tong SL (2013) Cohort profile: the China-Anhui Birth Cohort Study. Int J Epidemiol 42(3):709–721. https://doi.org/10.1093/ije/dys085

  32. 32.

    Matuszewski BK, Constanzer ML, Chavez-Eng CM (2003) Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC–MS/MS. Anal Chem 75(13):3019–3030. https://doi.org/10.1021/ac020361s

  33. 33.

    Andra SS, Austin C, Patel D, Dolios G, Awawda M, Arora M (2017) Trends in the application of high-resolution mass spectrometry for human biomonitoring: an analytical primer to studying the environmental chemical space of the human exposome. Environ Int 100:32–61. https://doi.org/10.1016/j.envint.2016.11.026

  34. 34.

    Dennis KK, Auerbach SS, Balshaw DM, Cui Y, Fallin MD, Smith MT, Spira A, Sumner S, Miller GW (2016) The importance of the biological impact of exposure to the concept of the exposome. Environ Health Perspect 124(10):1504–1510. https://doi.org/10.1289/EHP140

  35. 35.

    Wang H, Wang B, Zhao Q, Zhao Y, Fu C, Feng X, Wang N, Su M, Tang C, Jiang F, Zhou Y, Chen Y, Jiang Q (2015) Antibiotic body burden of Chinese school children: a multisite biomonitoring-based study. Environ Sci Technol 49(8):5070–5079. https://doi.org/10.1021/es5059428

  36. 36.

    Wang HX, Ren LS, Yu X, Hu J, Chen Y, He GS, Jiang QW (2017) Antibiotic residues in meat, milk and aquatic products in Shanghai and human exposure assessment. Food Control 80:217–225. https://doi.org/10.1016/j.foodcont.2017.04.034

  37. 37.

    Mokh S, El Hawari K, Nassar R, Budzinski H, Al Iskandarani M (2015) Optimization of a solid-phase extraction method for the determination of 12 aminoglycosides in water samples using LC–ESI–MS/MS. Chromatographia 78(9–10):631–640. https://doi.org/10.1007/s10337-015-2877-z

  38. 38.

    Lerbech AM, Opintan JA, Bekoe SO, Ahiabu MA, Tersbol BP, Hansen M, Brightson KT, Ametepeh S, Frimodt-Moller N, Styrishave B (2014) Antibiotic exposure in a low-income country: screening urine samples for presence of antibiotics and antibiotic resistance in coagulase negative staphylococcal contaminants. PLoS ONE 9(12):e113055. https://doi.org/10.1371/journal.pone.0113055

  39. 39.

    Zhu Y, Liu K, Zhang J, Liu X, Yang L, Wei R, Wang S, Zhang D, Xie S, Tao F (2020) Antibiotic body burden of elderly Chinese population and health risk assessment: a human biomonitoring-based study. Environ Pollut 256:113311. https://doi.org/10.1016/j.envpol.2019.113311

Download references

Acknowledgements

The authors thank Guanjun Chen and Shulong Li of the Center for Scientific Research of Anhui Medical University for valuable help in our experiment. We are deeply grateful for the help provided by all the members in the experimental center platform for physical and chemical of Anhui Medical University.

Funding

This work was supported by the National Natural Science Foundation of China (81202209), the Key Projects introduced and funded by leading talent teams of colleges and universities of Anhui province (0303011224) and Key Projects of Natural Science Research in Colleges and Universities of Anhui province (KJ2018A0164).

Author information

Correspondence to Fang-biao Tao.

Ethics declarations

Conflict of interest

The authors declare that there is no conflict regarding publication of this paper.

Ethical approval

All procedures involving human participants in this study were in accordance with the Helsinki declaration. This study was approved by the Anhui medical university human research ethics committee (Ethical approval code: 20131195).

Informed consent

Oral and written consents were acquired from all participants in the current study.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 2379 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Liu, K., Zhang, J., Geng, M. et al. A Stable Isotope Dilution Assay for Multi-class Antibiotics in Pregnant Urines by LC–MS/MS. Chromatographia (2020). https://doi.org/10.1007/s10337-020-03866-3

Download citation

Keywords

  • Liquid chromatography-triple quadrupole tandem mass spectrometry
  • Bio-monitoring
  • Antibiotics
  • China-Anhui Birth Cohort
  • Urine