Skip to main content
Log in

Novel Two-Dimensional Liquid Chromatography–Tandem Mass Spectrometry for the Analysis of Twenty Antibiotics Residues in Dairy Products

  • Published:
Food Analytical Methods Aims and scope Submit manuscript


A novel two-dimensional hydrophilic interaction chromatography/reversed phase liquid chromatography–tandem mass spectrometry system was developed to analyze seven categories of antibiotics residues in dairy products. The seven categories were β-lactams, tetracyclines, macrolides, aminoglycosides, amphenicols, quinolones, and sulphonamides, and involved 20 antibiotics. Samples of milk powder or milk samples were extracted by mixed matrix solid phase dispersion using C18 and CN material. The analytes were eluted by acetonitrile and water, and were dried with rotary evaporation. Finally, the residues were dissolved with mobile phase and then analyzed. Under sample pretreatment conditions, chromatographic and mass spectrometric parameters were optimized. Twenty analytes showed good linearities with correlation coefficients of 0.9945 ~ 0.9998. The limits of detection and quantification for milk powder and milk samples were 0.10 ~ 2.40 and 0.33 ~ 7.92 μg/kg, respectively. The proposed method has been applied to the determination of antibiotics residues in milk powder and milk samples.

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

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

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

Similar content being viewed by others


  • Aguilera-Luiz MM, Vidal JL, Romero-González R, Frenich AG (2008) Multi-residue determination of veterinary drugs in milk by ultra-high-pressure liquid chromatography-tandem mass spectrometry. J Chromatogr A 1205:10–16

    Article  CAS  Google Scholar 

  • Buszewski B, Noga S (2012) Hydrophilic interaction liquid chromatography (HILIC)—a powerful separation technique. Anal Bioanal Chem 402:231–247

    Article  CAS  Google Scholar 

  • Carretero V, Blasco C, Picó Y (2008) Multi-class determination of antimicrobials in meat by pressurized liquid extraction and liquid chromatography-tandem mass spectrometry. J Chromatogr A 1209:162–173

    Article  CAS  Google Scholar 

  • Chauve B, Guillarme D, Cléon P, Veuthey JL (2010) Evaluation of various HILIC materials for the fast separation of polar compounds. J Sep Sci 33:752–764

    Article  CAS  Google Scholar 

  • Chico J, Rúbies A, Centrich F, Companyó R, Prat MD, Granados M (2008) High-throughput multiclass method for antibiotic residue analysis by liquid chromatography-tandem mass spectrometry. J Chromatogr A 1213:189–199

    Article  CAS  Google Scholar 

  • Cristofani E, Antonini C, Tovo G, Fioroni L, Piersanti A, Galarini R (2009) A confirmatory method for the determination of tetracyclines in muscle using high-performance liquid chromatography with diode-array detection. Anal Chim Acta 637:40–46

    Article  CAS  Google Scholar 

  • Dejaegher B, Vander Heyden Y (2010) HILIC methods in pharmaceutical analysis. J Sep Sci 33:698–715

    Article  CAS  Google Scholar 

  • Fountain KJ, Xu J, Diehl DM, Morrison D (2010) Influence of stationary phase chemistry and mobile-phase composition on retention, selectivity, and MS response in hydrophilic interaction chromatography. J Sep Sci 33:740–751

    Article  CAS  Google Scholar 

  • Gentili A, Perret D, Marchese S (2005) Liquid chromatography-tandem mass spectrometry for performing confirmatory analysis of veterinary drugs in animal-food products. Trends Anal Chem 24:704–733

    Article  CAS  Google Scholar 

  • Gong Q, Ding L, Zhu S, Jiao Y, Cheng J, Fu S, Wang L (2012) Determination of ten aminoglycoside residues in milk and dairy products using high performance liquid chromatography-tandem mass spectrometry. Chinese J Chromatography 30:1143–1147

    Article  CAS  Google Scholar 

  • Goucher E, Kicman A, Wolff K, Smith N, Jickells S (2010) Hydrophilic stationary phases: a practical approach for the co-analysis of compounds with varying polarity in biological matrices. J Sep Sci 33:955–965

    Article  CAS  Google Scholar 

  • Granelli K, Branzell C (2007) Rapid multi-residue screening of antibiotics in muscle and kidney by liquid chromatography-electrospray ionization-tandem mass spectrometry. Anal Chim Acta 586:289–295

    Article  CAS  Google Scholar 

  • Granelli K, Elgerud C, Lundström A, Ohlsson A, Sjöberg P (2009) Rapid multi-residue analysis of antibiotics in muscle by liquid chromatography-tandem mass spectrometry. Anal Chim Acta 637:87–91

    Article  CAS  Google Scholar 

  • Grumbach ES, Diehl DM, Neue UD (2008) The application of novel 1.7 microm ethylene bridged hybrid particles for hydrophilic interaction chromatography. J Sep Sci 31:1511–1518

    Article  CAS  Google Scholar 

  • Hemström P, Irgum K (2006) Hydrophilic interaction chromatography. J Sep Sci 29:1784–1821

    Article  Google Scholar 

  • Jandera P (2008) Stationary phases for hydrophilic interaction chromatography, their characterization and implementation into multidimensional chromatography concepts. J Sep Sci 31:1421–1437

    Article  CAS  Google Scholar 

  • Jandera P, Hájek T, Skeríková V, Soukup J (2010) Dual hydrophilic interaction-RP retention mechanism on polar columns: structural correlations and implementation for 2-D separations on a single column. J Sep Sci 33:841–852

    Article  CAS  Google Scholar 

  • Jandera P, Hájek T, Staňková M, Vyňuchalová K, Česla P (2012) Optimization of comprehensive two-dimensional gradient chromatography coupling in-line hydrophilic interaction and reversed phase liquid chromatography. J Chromatogr A 1268:91–101

    Article  CAS  Google Scholar 

  • Kahsay G, Song H, Van Schepdael A, Cabooter D, Adams E (2014) Hydrophilic interaction chromatography (HILIC) in the analysis of antibiotics. J Pharm Biomed Anal 87:142–154

    Article  CAS  Google Scholar 

  • Kennedy DG, McCracken RJ, Cannavan A, Hewitt SA (1998) Use of liquid chromatography-mass spectrometry in the analysis of residues of antibiotics in meat and milk. J Chromatogr A 812:77–98

    Article  CAS  Google Scholar 

  • Kumar P, Rubies A, Companyó R, Centrich F (2012) Hydrophilic interaction chromatography for the analysis of aminoglycosides. J Sep Sci 35:498–504

    Article  CAS  Google Scholar 

  • Liu M, Chen EX, Ji R, Semin D (2008a) Stability-indicating hydrophilic interaction liquid chromatography method for highly polar and basic compounds. J Chromatogr A 1188:255–263

    Article  CAS  Google Scholar 

  • Liu Y, Xue X, Guo Z, Xu Q, Zhang F, Liang X (2008b) Novel two-dimensional reversed-phase liquid chromatography/hydrophilic interaction chromatography, an excellent orthogonal system for practical analysis. J Chromatogr A 1208:133–140

    Article  CAS  Google Scholar 

  • Nakazawa H, Ino S, Kato K, Watanabe T, Ito Y, Oka H (1999) Simultaneous determination of residual tetracyclines in foods by high-performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry. J Chromatogr B Biomed Sci Appl 732:55–64

    Article  CAS  Google Scholar 

  • Nguyen HP, Yang SH, Wigginton JG, Simpkins JW, Schug KA (2010) Retention behavior of estrogen metabolites on hydrophilic interaction chromatography stationary phases. J Sep Sci 33:793–802

    Article  CAS  Google Scholar 

  • Nováková L, Kaufmannová I, Jánská R (2010) Evaluation of hybrid hydrophilic interaction chromatography stationary phases for ultra-HPLC in analysis of polar pteridines. J Sep Sci 33:765–772

    Article  Google Scholar 

  • van Holthoon FL, Essers ML, Mulder PJ, Stead SL, Caldow M, Ashwin HM, Sharman M (2009) A generic method for the quantitative analysis of aminoglycosides (and spectinomycin) in animal tissue using methylated internal standards and liquid chromatography tandem mass spectrometry. Anal Chim Acta 637:135–143

    Article  Google Scholar 

  • Wang L, Li YQ, Wang HB, Guan YL (2011) Simultaneous determination of twenty veterinary drug residues in milk and meat using matrix solid phase dispersion-ultra performance liquid chromatography-tandem mass spectrometry. Chinese Journal of Anal Chem 39:203–207

    Google Scholar 

  • Wu J, Bicker W, Lindner W (2008) Separation properties of novel and commercial polar stationary phases in hydrophilic interaction and reversed-phase liquid chromatography mode. J Sep Sci 31:1492–1503

    Article  CAS  Google Scholar 

  • Zurhelle G, Müller-Seitz E, Petz M (2000) Automated residue analysis of tetracyclines and their metabolites in whole egg, egg white, egg yolk and hen's plasma utilizing a modified ASTED system. J Chromatogr B Biomed Sci Appl 739:191–203

    Article  CAS  Google Scholar 

Download references


This study was funded by the China Postdoctoral Science Foundation (2012M521703). We thank Prof. Dongtao Lin of the Sichuan University for copyediting the manuscript.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Lian Wang.

Ethics declarations

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Conflict of Interest

Lian Wang declares that he has no conflict of interest, Bixia Yang declares that she has no conflict of interest, Xinshen Zhang declares that he has no conflict of interest, and Hongguo Zheng declares that he has no conflict of interest.

Informed Consent

Not applicable.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, L., Yang, B., Zhang, X. et al. Novel Two-Dimensional Liquid Chromatography–Tandem Mass Spectrometry for the Analysis of Twenty Antibiotics Residues in Dairy Products. Food Anal. Methods 10, 2001–2010 (2017).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: