Skip to main content
SpringerLink
Log in

Separation and Characterization of New Components and Impurities in Leucomycin by Multiple Heart-Cutting Two-Dimensional Liquid Chromatography Combined with Ion Trap/Time-of-Flight Mass Spectrometry

  • Original
  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

In this study, nine new components and six impurities in leucomycin were discovered. A method was developed for the separation and characterization of new components and impurities in leucomycin by multiple heart-cutting two-dimensional liquid chromatography combined with ion trap/time-of-flight mass spectrometry in both positive and negative electrospray ionization modes. With this method, a non-volatile buffer solution was used as mobile phase in the first-dimensional system for good separation. Eluent of each peak from the first-dimensional system was trapped by a switching valve and sent to the liquid chromatography-mass spectrometry system using a volatile mobile phase. The complete fragmentation patterns of the new components and degradation impurities were deduced based on MSn data. The structures of nine new components in leucomycin were deduced as unsaturated ketone in the 16-membered ring of leucomycin. The structures of six impurities were characterized for the first time, four of which were acid degradation products, and the other two were process impurities. The correlation between impurities and the purification process of leucomycin was also studied. The degradation impurities were produced during purification of leucomycin fermentation broth, which requires a low-pH environment. Based on the characterization of impurities, this study not only revealed the mechanism of impurity production, thus providing guidance to pharmaceutical companies for manufacturing process improvement and impurity reduction, but also provided a scientific basis for further improvement of official monographs in pharmacopoeias.

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
Fig. 6

Similar content being viewed by others

References

  1. Liu QQ (2003) Improved preparation of kitasamycin tablets. Chin J Pharm 34:22–23

    Google Scholar 

  2. Yang Q, Ma SH, Hu M, Hu CQ (2006) Determination of active ingredients of kitasamycin by high performance liquid chromatography. Chin J Anal Chem 34:95–99

    CAS  Google Scholar 

  3. Balducci Y, Balducci Y, Bodey GP, Bodey GP (1974) In vitro activity of kitasamycin against gram-positive cocci. J Antibiot 27:516–519

    Article  CAS  PubMed  Google Scholar 

  4. Filadoro F, Cipriani P, Ravagnan L (1968) Antibacterial effect in vitro and in vivo of kitasamycin. Antibiotica 6:5–23

    CAS  PubMed  Google Scholar 

  5. ICH Harmonised Tripartite Guideline Q3A(R) (2006) Impurities in new salt substances, The International Council for Harmonisation (ICH) of Technical Requirements for Pharmaceuticals for Human Use. https://www.ich.org/products/guidelines/quality/article/quality-guidelines.html. Accessed 25 Jan 2019

  6. Fukutsu N, Kawasaki T, Saito K, Nakazawa H (2006) Application of high-performance liquid chromatography hyphenated techniques for identification of degradation products of cefpodoxime proxetil. J Chromatogr A 1129:153–159

    Article  CAS  PubMed  Google Scholar 

  7. Pan YH, Zhang HY, Xi CG, Huang LL, Xie SY, Chen DM, Tao YF, Liu ZL, Yuan ZH (2018) Simultaneous determination of multicomponent of acetylkitasamycin and kitasamycin by LC-MS/MS in swine plasma and its application in a pharmacokinetic study. Biomed Chromatogr 32:e4268

    Article  CAS  Google Scholar 

  8. Chen X (2011) Study on contents determination methodology of kitasamycinum composition in kitasamycin tartrate for injection by HPLC. J Hubei Univ Med 30:588–590

    Google Scholar 

  9. Hu M, Hu CQ (2006) Identification of the components and products of hydrolysis in acetyl leucomycin by LC-MS. Acta Pharm Sin 41:476–480

    CAS  Google Scholar 

  10. Zhu SQ, Niu CQ (2007) LC-MS analysis of components of kitasamycin. Chin J Antibiot 32:478–480

    CAS  Google Scholar 

  11. Van den Bossche F, Daidone F, Van Schepdael A, Hoogmartens J, Adams E (2013) Characterization of impurities in josamycin using dual liquid chromatography combined with mass spectrometry. J Pharm Biomed Anal 73:66–76

    Article  CAS  PubMed  Google Scholar 

  12. Wang MJ, Wang Y, Li J, Li YP, Hu CQ, Hoogmartens J, Van Schepdael A, Adams E (2013) Characterization of the components of meleumycin by liquid chromatography with photo-diode array detection and electrospray ionization tandem mass spectrometry. J Pharm Biomed Anal 84:69–76

    Article  CAS  PubMed  Google Scholar 

  13. Wang MJ, Hu CQ (2013) Impurity profiling of macrolide antibiotics by liquid chromatography-mass spectrometry. Acta Parm Sin 48:642–647

    CAS  Google Scholar 

  14. Holm R, Elder DP (2016) Analytical advances in pharmaceutical impurity profiling. Eur J Pharm Sci 87:118–135

    Article  CAS  PubMed  Google Scholar 

  15. Martano C, Ferretti F, Ghiani S, Buonsanti F, Bruno E, Lattuada L, Medana C (2017) Development and validation of a new HPLC-MS method for meglumine impurity profiling. J Pharm Biomed Anal 149:517–524

    Article  CAS  PubMed  Google Scholar 

  16. Shipkova PA, Heimark L, Bartner PL, Chen G, Pramanik BN, Ganguly AK, Cody RB, Kusai A (2000) High-resolution LC/MS for analysis of minor components in complex mixtures: negative ion ESI for identification of impurities and degradation products of a new oligosaccharide antibiotic. J Mass Spectrom 35:1252–1258

    Article  CAS  PubMed  Google Scholar 

  17. Reig MN, Jaumot J, Baglai A, Truyols GV, Schoenmakers PJ, Tauler R (2017) Untargeted comprehensive two-dimensional liquid chromatography coupled with high-resolution mass spectrometry analysis of rice metabolome using multivariate curve resolution. Anal Chem 89:7675–7683

    Article  CAS  Google Scholar 

  18. Yang Q, Wang ZY, Tang SF (2016) Application of two-dimensional UPLC-QTof MS technology in the study of the impurity profile of bleomycin hydrochloride*. Chin J Pharm Anal 36:1231–1242

    Google Scholar 

  19. Long Z, Zhan ZQ, Guo ZM, Li YQ, Li CK, Yao JT, Ji F, Zheng X, Ren B, Huang TH (2019) A novel two-dimensional liquid chromatography-mass spectrometry method for direct drug impurity identification from HPLC eluent containing ion-pairing reagent in mobile phases. Anal Chim Acta 1049:105–114

    Article  CAS  PubMed  Google Scholar 

  20. Wang J, Xu Y, Wen C, Wang Z (2017) Application of a trap-free two-dimensional liquid chromatography combined with ion trap/time-of-flight mass spectrometry for separation and characterization of impurities and isomers in cefpiramide. Anal Chim Acta 992:42–54

    Article  CAS  PubMed  Google Scholar 

  21. Petersson P, Haselmann K, Buckenmaier S (2016) Multiple heart-cutting two dimensional liquid chromatography mass spectrometry: towards real time determination of related impurities of bio-pharmaceuticals in salt based separation methods. J Chromatogr A 1468:95–101

    Article  CAS  PubMed  Google Scholar 

  22. Schans MGMVD, Blokland MH, Zoontjes PW, Mulder PPJ, Nielen MWF (2017) Multiple heart-cutting two dimensional liquid chromatography quadrupole time-of-flight mass spectrometry of pyrrolizidine alkaloids. J Chromatogr A 1503:38–48

    Article  CAS  PubMed  Google Scholar 

  23. National Pharmacopoeia Committee (2015) Chinese pharmacopoeia, part 2, 2015th edn. China Medical Science and Technology Press, Beijing

    Google Scholar 

  24. Editorial Board of Japanese Pharmaceutical (2016) The Japanese pharmacopoeia, Seventeenth edn. Ministry of Health, Labour and Welfare

    Google Scholar 

  25. Govaerts C, Chepkwony HK, Van Schepdael A, Adams E, Roets E, Hoogmartens J (2004) Application of liquid chromatography-ion trap mass spectrometry to the characterization of the 16-membered ring macrolide josamycin propionate. J Mass Spectrom 39:437–446

    Article  CAS  PubMed  Google Scholar 

  26. Hu M, Hu CQ (2005) Identification of the components of 16-membered macrolide antibiotics by LC/MS. Anal Chim Acta 535:89–99

    Article  CAS  Google Scholar 

  27. Przybylski P, Pyta K, Brzezinski B (2010) Fragmentation pathways of new aza derivatives of 16-membered macrolide antibiotic-analog of Josamycin investigated by ESI and FAB mass spectrometric methods. J Mass Spectrom 44:1395–1401

    Article  CAS  Google Scholar 

  28. Zhang X, Li J, Wang C, Song DQ, Hu CQ (2017) Identification of impurities in macrolides by liquid chromatography–mass spectrometric detection and prediction of retention times of impurities by constructing quantitative structure–retention relationship (QSRR). J Pharm Biomed Anal 145:262–272

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by Key Technologies and Standards for Drug Consistency Assessment of National Science and Technology Major Project (No. 2017ZX09101001).

Author information

Authors and Affiliations

  1. Zhejiang University of Technology, Hangzhou, 310014, China

    Jian Wang, Guijun Liu & Yu Xu

  2. Zhejiang Institute for Food and Drug Control, Hangzhou, 310052, China

    Jian Wang, Bingqi Zhu & Zhijian Wang

Authors
  1. Jian Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guijun Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yu Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bingqi Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhijian Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhijian Wang.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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

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 material 1 (DOC 2724 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, J., Liu, G., Xu, Y. et al. Separation and Characterization of New Components and Impurities in Leucomycin by Multiple Heart-Cutting Two-Dimensional Liquid Chromatography Combined with Ion Trap/Time-of-Flight Mass Spectrometry. Chromatographia 82, 1333–1344 (2019). https://doi.org/10.1007/s10337-019-03754-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10337-019-03754-5

Keywords

Search

Navigation