, Volume 81, Issue 2, pp 303–314 | Cite as

Simultaneous Screening and Determination of Eight Tetracycline Antibiotics Illegally Adulterated in Herbal Preparations Using HPLC–DAD Combined with LC–MS–MS

  • Fuyan Liu
  • Xiaofeng Zhang
  • Yuqiu Li
  • Hui Gao
  • Peixue Ling
  • Xiaoyan Li
  • Qixin Chen
  • Aibin Ma
  • Huarong Shao
  • Mei Li
  • Fengshan Wang


In this study, the applicability of high-performance liquid chromatography with diode-array detection (HPLC–DAD) combined with electrospray ionization tandem mass spectrometry (ESI–MS) for the simultaneous screening and determination of tetracycline antibiotic analogues illegally adulterated in herbal preparations was investigated. The HPLC–DAD method could be employed to separate and simultaneously determine eight tetracyclines using an isocratic solvent system. These tetracycline antibiotics include doxycycline, 6-epidoxycycline and tetracycline, which are difficult to identify because they are structural isomers. ESI–MS was used in combination with HPLC–DAD to improve the qualitative accuracy in this study. Method validation included the investigation of linearity, selectivity, stability, limits of detection and quantitation, trueness, precision and ruggedness. The response exhibited a good linear relationship with the analyte concentration. The coefficient of determination (r 2) was greater than 0.9993. The limits of quantification for the examined tetracycline antibiotics ranged from 1.11 to 25.73 µg·mL−1, and the recoveries ranged from 91.0 to 104.0%. The proposed methods were specific, sensitive and accurate, and they could be promising and powerful tools for the routine screening of tetracycline analogues in herbal medicines to ensure food safety and public health.

Graphical abstract


Illegal adulteration Tetracycline antibiotics HPLC–DAD LC/MS/MS Herbal preparations Screening analysis 









Ethylenediamine tetraacetic acid


Electrospray ionization tandem mass spectrometry




High-performance liquid chromatography with diode-array detection


Limit of detection


Limit of quantitation








The relative standard deviation


Retention time





This work was supported by the Shandong Public Service Platform for new drug safety and pharmacology evaluation, Shandong Provincial Research Fund for Outstanding Young Scholars (BS2015YY037).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

10337_2017_3450_MOESM1_ESM.docx (328 kb)
Supplementary material 1 (DOCX 327 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  1. 1.School of Pharmaceutical SciencesShandong UniversityJinanChina
  2. 2.Shandong Provincial Key Laboratory of BiopharmaceuticalsShandong Academy of Pharmaceutical ScienceJinanChina
  3. 3.Shandong Institute for Food and Drug ControlJinanChina
  4. 4.School of Life SciencesLanzhou UniversityLanzhouChina
  5. 5.Shandong University of Traditional Chinese MedicineJinanChina

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