Skip to main content

Gas Chromatography–Mass Spectrometry Determination of Pregabalin in Human Plasma Using Derivatization Method


A gas chromatography–mass spectrometry method for the determination of pregabalin in human plasma is described. The procedure involves precipitation of protein, liquid–liquid extraction with ethylene glycol monomethyl ether, and derivatization with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide in the presence of N-hydroxysuccinimide as additive. Separation was attained on HP column (30 m × 0. 25 mm ID, 0.25 μm) coupled with mass spectrometric detector using electron impact selected ion monitoring. The assay showed an excellent linearity in the concentration range of 0.36–10 μg mL−1 with correlation coefficient (r2) values of 0.999. The intra- and inter-day assay variations for three different concentration levels were less than 10%. The limit of quantification was detected at 0.36 μg mL−1. The method is highly specific, precise, accurate, and reproducible and could also be applied for the determination of pregabalin in human plasma.

This is a preview of subscription content, access via your institution.

Fig. 1
Scheme 1
Fig. 2
Fig. 3
Fig. 4


  1. Ben-Menachem E (2004) Pregabalin pharmacology and its relevance to clinical practice. Epilepsia 45:13–18

    Article  CAS  Google Scholar 

  2. Finnerup NB, Jensen TS (2007) Clinical use of pregabalin in the management of central neuropathic pain. Neuropsychiatr Dis Treat 3:885–891

    Article  CAS  Google Scholar 

  3. Tzellos TG, Papazisis G, Amaniti E, Kouvelas D (2008) Efficacy of pregabalin and gabapentin for neuropathic pain in spinal-cord injury: an evidence-based evaluation of the literature. ‎Eur J Clin Pharmacol 64:851–858

    Article  CAS  Google Scholar 

  4. Ahmadkhaniha R, Mottaghi S, Zargarpoor M, Souri E (2014) Validated HPLC method for quantification of pregabalin in human plasma using 1-Fluoro-2,4-dinitrobenzene as derivatization agent. Chromatogr Res Int 2014:450461

    Article  Google Scholar 

  5. Berry D, Millington C (2005) Analysis of pregabalin at therapeutic concentrations in human plasma/serum by reversed-phase HPLC. Ther Drug Monit 27:451–456

    Article  CAS  Google Scholar 

  6. Gujral RS, Haque SM, Kumar S (2009) A novel method for the determination of pregabalin in bulk pharmaceutical formulations and human urine samples. Afr J Pharm Pharmacol 3:327–334

    CAS  Google Scholar 

  7. Arayne MS, Shahnaz H, Ali A, Sultana N (2014) Monitoring of pregabalin in pharmaceutical formulations and human serum using UV and RP-HPLC techniques: application to dissolution test method. Pharma Anal Acta 5:287–293

    Article  Google Scholar 

  8. Gujral RS, Haque SM, Shanker P (2009) A sensitive spectrophotometric method for the determination of pregabalin in bulk, pharmaceutical formulations and in human urine samples. Int J Biomed Sci 5:421–427

    CAS  Google Scholar 

  9. Shaalan RA (2010) Spectrofluorimetric and spectrophotometric determination of pregabalin in capsules and urine samples. Int J Biomed Sci 6:260–267

    CAS  Google Scholar 

  10. Jang K, Seo J, Yim S, Lee K (2011) Rapid and simple method for the determination of pregabalin in human plasma using liquid chromatography–tandem mass spectrometry (LC–MS/MS): application to a bioequivalence study of Daewoong Pregabalin capsule to Lyrica® Capsule (Pregabalin 150 mg). Int J Pharm Investig 41:255–262

    Article  CAS  Google Scholar 

  11. Mandal U, Kanti Sarkar A, Veeran Gowda K, Agarwal S, Bose A, Bhaumik U, Ghosh D, Kumar Pal T (2008) Determination of pregabalin in human plasma using LC–MS–MS. Chromatographia 67:237–243

    Article  CAS  Google Scholar 

  12. Nirogi R, Kandikere V, Mudigonda K, Komarneni P, Aleti R (2009) Liquid chromatography atmospheric pressure chemical ionization tandem mass spectrometry method for the quantification of pregabalin in human plasma. J Chromatogr B 877:3899–3906

    Article  CAS  Google Scholar 

  13. Thejaswini JC, Gurupadayya BM, Raja P (2012) Gas chromatographic determination of pregabalin in human plasma using ethyl chloroformate derivatizing reagent. J Pharm Res 5:3112–3115

    CAS  Google Scholar 

  14. Aji A, Karthikeyan S, Singh S, Puthli SP (2013) A novel UPLC–MS/MS method for determination of γ-amino butyric acid analogue in human Plasma: application to pharmacokinetic study. J Appl Pharm Sci 3:172–178

    Google Scholar 

  15. Mudiam MKR, Chauhan A, Jain R, Ratnasekhar C, Fatima G, Malhotra E, Murthy RC (2013) Development, validation and comparison of two microextraction techniques for the rapid and sensitive determination of pregabalin in urine and pharmaceutical formulations after ethyl chloroformate derivatization followed by gas chromatography–mass spectrometric analysis. J Pharm Biomed Anal 70:310–319

    Article  Google Scholar 

  16. Goodarzi N, Ghahremani MH, Amini M, Atyabi F, Ostad SN, Shabani Ravari N, Nateghian N, Dinarvand R (2014) CD44-targeted docetaxel conjugate for cancer cells and cancer stem-like cells: a novel hyaluronic acid-based drug delivery system. Chem Biol Drug Des 83:741–752

    Article  CAS  Google Scholar 

  17. Dinan NM, Atyabi F, Rouini MR, Amini M, Golabchifar AA, Dinarvand R (2014) Doxorubicin loaded folate-targeted carbon nanotubes: preparation, cellular internalization, in vitro cytotoxicity and disposition kinetic study in the isolated perfused rat liver. Mater Sci Eng C 39:47–55

    Article  CAS  Google Scholar 

  18. Amini M, Navidpour L, Shafiee A (2014) Synthesis and antitubercular activity of new N,N-diaryl-4-(4,5-dichloroimidazole-2-yl)-1,4-dihydro-2,6-dimethyl-3,5-pyridinedicarboxamides. Daru 16:9–12

    Google Scholar 

  19. Daman Z, Ostad S, Amini M, Gilani K (2014) Preparation, optimization and in vitro characterization of stearoyl-gemcitabine polymeric micelles: a comparison with its self-assembled nanoparticles. Int J Pharm 468:142–152

    Article  CAS  Google Scholar 

  20. Figueiredo RM, Suppo JS, Campagne JM (2016) Nonclassical routes for amide bond formation. Chem Rev 116:12029–12122

    Article  Google Scholar 

  21. Food and Drug Administration of the United States (2001) Guidance for industry-Bioanalytical Method Validation, U.S. Department of Health and Human Services, Center for Drug Evaluation and Research (CDER), Center for Veterinary Medicine (CVM)

  22. Arroyo S, Anhut H, Kugler AR, Lee CM, Knapp LE, Garofalo EA, Messmer S (2004) Pregabalin add-on treatment: a randomized, double-blind, placebo-controlled, dose–response study in adults with partial seizures. Epilepsia 45:20–27

    Article  CAS  Google Scholar 

Download references


The authors acknowledged Food and drug Administration and Drug Design and Development Research Center, Tehran University of Medical Sciences, Tehran, Iran, for their support by providing the necessary equipment’s and chemicals so as to complete the study. The current work was financially supported by Tehran University of Medical Sciences.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Mohsen Amini.

Ethics declarations

Conflict of interest

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

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Rights and permissions

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tafesse, T.B., Mazdeh, F.Z., Chalipour, A. et al. Gas Chromatography–Mass Spectrometry Determination of Pregabalin in Human Plasma Using Derivatization Method. Chromatographia 81, 501–508 (2018).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: