Skip to main content
Log in

Quantification of free polyamines and their metabolites in biofluids and liver tissue by UHPLC-MS/MS: application to identify the potential biomarkers of hepatocellular carcinoma

  • Note
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript


Polyamines and their N-acetylated metabolites are potential biomarkers in the diagnosis and therapeutic evaluation of cancer. Thus, we present here an ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous determination of 6 free, 3 monoacetylated, and 2 diacetylated polyamines without derivatization. The major improvement of this method is the use of 0.2 % perfluoroheptanoic acid methanol in the pretreatment step to achieve protein precipitation and 0.0125 % perfluoroheptanoic acid in the mobile phase to achieve analyte separation within 9 min. The established analytical method was validated with plasma, urine, and liver tissue and applied to determine plasma, urine, and liver tissue samples from healthy rats, hepatocellular carcinoma rats, and administrated rats successfully. Results indicated free polyamines such as putrescine mainly existed in liver tissue but more polar N-acetylated metabolites such as N 1,N 12-diacetylspermine seemed to exist in biological fluid. After carcinogenesis, the levels of polyamines were increased, but the elevated levels of polyamines and their metabolites tended to decrease when administrated with anticancer drug. The method provided a more versatile manner for clinical application in the diagnosis and therapeutic evaluation of hepatocellular carcinoma.

The process of quantification of free polyamines and their metabolites in biofluids and liver tissue by UHPLC-MS/MSᅟ

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others


  1. Goyal L, Supko JG, Berlin J, Blaszkowsky LS, Carpenter A, Heuman DM, Hilderbrand SL, Stuart KE, Cotler S, Senzer NN, Chan E, Berg CL, Clark JW, Hezel AF, Ryan DP, Zhu A (2013) Phase 1 study of N 1,N 11-diethylnorspermine (DENSPM) in patients with advanced hepatocellular carcinoma. Cancer Chemoth Pharm 72:1305–1314

    Article  CAS  Google Scholar 

  2. Tan GS, Lim KH, Tan HT, Khoo ML, Tan SH, Toh HC, Ming C, Chung M (2014) Novel proteomic biomarker panel for prediction of aggressive metastatic hepatocellular carcinoma relapse in surgically resectable patients. J Proteome Res 13(11):4833–4846

    Article  CAS  Google Scholar 

  3. Moinard C, Cynober L, de Bandt JP (2005) Polyamines: metabolism and implications in human diseases. Clin Nutr 24(2):184–197

    Article  CAS  Google Scholar 

  4. Russell DH (1971) Increased polyamine concentrations in the urine of human cancer patients. Nat New Biol 233(39):144–145

    Article  CAS  Google Scholar 

  5. Samejima K, Hiramatsu K, Takahashi K, Kawakita M, Kobayashi M, Tsumoto H, Kohda K (2010) Identification and determination of urinary acetylpolyamines in cancer patients by electrospray ionization and time-of-flight mass spectrometry. Anal Biochem 401(1):22–29

    Article  CAS  Google Scholar 

  6. Gaboriau F, Havouis R, Moulinoux JP, Delcros JG (2003) Atmospheric pressure chemical ionization-mass spectrometry method to improve the determination of dansylated polyamines. Anal Biochem 318(2):212–220

    Article  CAS  Google Scholar 

  7. Häkkinen MR (2011) Polyamine analysis by LC-MS. Methods Mol Biol 720:505–518

    Article  Google Scholar 

  8. Feistner GJ (1994) Profiling of basic amino acids and polyamines in microbial culture supernatants by electrospray mass spectrometry. Biol Mass Spectrom 23(12):784–792

    Article  CAS  Google Scholar 

  9. Liu R, Li Q, Ma R, Lin X, Xu H, Bi K (2013) Determination of polyamine metabolome in plasma and urine by ultrahigh performance liquid chromatography–tandem mass spectrometry method: application to identify potential markers for human hepatic cancer. Anal Chim Acta 791:36–45

    Article  CAS  Google Scholar 

  10. Häkkinen MR, Roine A, Auriola S, Veskimäe E, Keinänen TA, Lehtimäki T, Oksala N, Vepsäläinen J (2013) Analysis of free, mono-and diacetylated polyamines from human urine by LC–MS/MS. J Chromatogr, B 941:81–89

    Article  Google Scholar 

  11. Sánchez-López J, Camañes G, Flors V, Vicent C, Pastor V, Vicedo B, Cerezo M, García-Agustín P (2009) Underivatized polyamine analysis in plant samples by ion pair LC coupled with electrospray tandem mass spectrometry. Plant Physiol Bioch 47(7):592–598

    Article  Google Scholar 

Download references


This study was financially supported by the National Natural Science Funds of China (grant No. 81102784/H2803), the key project in scientific research from the Ministry of Education (grant No. 202032), and Liaoning Innovative Research Team in university (LNIRT No. LT2013022).

Author information

Authors and Affiliations


Corresponding author

Correspondence to Qing Li.

Electronic supplementary material

Below is the link to the electronic supplementary material.


(PDF 3611 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yu, C., Liu, R., Xie, C. et al. Quantification of free polyamines and their metabolites in biofluids and liver tissue by UHPLC-MS/MS: application to identify the potential biomarkers of hepatocellular carcinoma. Anal Bioanal Chem 407, 6891–6897 (2015).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: