Acta Scientific Pharmaceutical Sciences (ISSN: 2581-5423)

Review ArticleVolume 5 Issue 4

A Review on Analytical methods: Levetiracetam

Malla Krishna Prasad* and Mukthinuthalapati Mathrusri Annapurna

Department of Pharmaceutical Analysis and Quality Assurance, GITAM Institute of Pharmacy, GITAM (Deemed to be University), Visakhapatnam, Andhra Pradesh, India

*Corresponding Author: Malla Krishna Prasad, Department of Pharmaceutical Analysis and Quality Assurance, GITAM Institute of Pharmacy, GITAM (Deemed to be University), Visakhapatnam, Andhra Pradesh, India.

Received: February 24, 2021; Published: March 09, 2021

Citation: Malla Krishna Prasad and Mukthinuthalapati Mathrusri Annapurna . “A Review on Analytical methods: Levetiracetam”. Acta Scientific Pharmaceutical Sciences 5.4 (2021): 03-05.


  Levetiracetam is a second-generation anti-epileptic drug which belongs to pyrrolidone family with wide spectrum of action. In 1999 Levetiracetam was approved by US Food and Drug Administration as a broad spectrum antiepileptic drug. Levetiracetam is different from other older antiepileptic drugs in its structure with hydrophilic groups. The present review summarizes the analytical techniques used for the analysis of Levetiracetam.

Keywords: Levetiracetam; Antiepileptic Drug; Spectrophotometry; HPLC; LC-MS


  Levetiracetam is an anti-epileptic agent [1] and chemically known as (S)-2-(2-oxopyrrolidin-1-yl) butanamide (Figure 1) and it has a molecular formula (C8H14N2O2). Levetiracetam is very soluble in water and freely soluble in methanol and chloroform. In n-hexane Levetiracetam is insoluble practically. Levetiracetam acts by modulating the synaptic neurotransmitter release and by binding itself to synaptic vesicle protein SV2A in the brain. Levetiracetam was approved by the US FDA. Levetiracetam is absorbed completely after the oral administration and about 100% bioavailability was reported. Levetiracetam undergoes metabolism through enzymatic hydrolysis of acetamide group. Levetiracetam maintains a large margin of safety and no interactions were reported with other anticonvulsants [2] and due to this Levetiracetam is used as an adjunctive therapy for the treatment of epileptic seizures.

Figure 1: Structure of Levetiracetam.

  The authors have summarised the analytical methods developed for the quantification of Levetiracetam in the present review article. These analytical methods include spectrophotometry [3-6], HPLC [7-12], UPLC [13], UHPLC [14], HPTLC [15], LC-MS/MS [16,17] and UPLC-MS/MS [18] for the estimation of Levetiracetam in pharmaceutical formulations and biological fluids such as human plasma and human serum (Table 1).

Reagent/Mobile phase (v/v)

λ (nm)





Spectrophotometric methods






2-Chloro phenyl hydrazine and

2 ml (0.25 %) Anthranilic acid






2,4- Dinitrophenylhydrazine





Glacial acetic acid





Liquid chromatographic methods

Potassium dihydrogen phosphate and sodium 1-heptane sulphonate buffer (pH adjusted to 2.8 with ortho phosphoric acid): Acetonitrile (90:10)





Acetonitrile: water

(Gradient mode) Internal standard




Human serum


Acetonitrile: 0.03 M potassium dihydrogen phosphate (pH adjusted to 3.0 with ortho phosphoric acid) (15: 85)





0.05M KH2PO4 buffer (pH adjusted to 3.0 with ortho phosphoric acid): Methanol (70:30)





Water: acetonitrile (90:10)





Methanol: Ammonium acetate buffer (pH 4.0) (80:20)

Ritonavir (Internal standard)





Buffer (KH2PO4 + Heptan sulphonic acid salt) (pH 2.4): Acetonitrile (90: 10)






Acetonitrile: water (80: 20) Lamivudine (Internal standard)




Human serum


Toluene: Ethyl acetate: Methanol (2:1:1)





Liquid chromatography-Mass spectrometric methods

5 mM Ammonium acetate (adjusted to pH 3.2 with Glacial acetic acid): Acetonitrile (20: 80) Clonazepam (Internal standard)



LC-MS/MS Human plasma


0.1% Formic acid in Ammonium acetate: Methanol (10: 90) Lamivudine (Internal standard)



LC-MS/MS Human plasma


[0.1%formic acid -10 mM Ammonium formate (pH 3.5)]:

[0.1%formic acid in methanol] (Gradient mode)




Human plasma


Table 1: Review of analytical methods for the determination of Levetiracetam.


  Different analytical methods such as UV, HPLC, HPTLC, UPLC, UHPLC and hyphenated techniques such as GC-MS, LC-MS methods were reported for the estimation of Levetiracetam in bulk, pharmaceutical formulations and biological samples. This review article is very much useful for the readers for understanding the analytical techniques so far reported for the quantification of Levetiracetam.


  1. The complete drug reference. 36th edition. Pharmaceutical press, London. (2009): 488-489.
  2. Yi ZM., et al. “Levetiracetam for epilepsy: an evidence map of efficacy, safety and economic profiles”. Neuropsychiatric Disease and Treatment 15 (2018): 1-19. 
  3. Ravisankar P., et al. “A simple validated UV spectrophotometric method for quantitative analysis of Levetiracetam in pharmaceutical dosage form”. Indian Journal of Research in Pharmacy and Biotechnology5 (2015): 380-385.
  4. Muralikrishna Ch., et al. “Spectrophotometric determination of levetiracetam by developing coloured complexes with 2-chlorophenylhydrazine and anthranilic acid”. Asian Journal of Chemistry 4 (2012): 1855-1857.
  5. Thanuja S., et al. “Spectrophotometric determination of Levetiracetam using 2, 4-DNP in pharmaceutical dosage form”. Indo American Journal of Pharmaceutical Research 4 (2014): 561-565.
  6. Madhu M., et al. “Development and validation of spectroscopic method for estimation of Levetiracetam in tablet dosage form”. Journal of Global Trends in Pharmaceutical Sciences4 (2015): 2956-2962.
  7. Valarmathy J., et al. “RP-HPLC method development and validation for assay of Levetiracetam in tablet dosage form”. Research Journal of Pharmacy and Technology3 (2008): 395-397.
  8. Vermeij TA., et al. “High-performance liquid chromatographic and megabore gas-liquid chromatographic determination of Levetiracetam (ucb L059) in human serum after solid-phase extraction”. Journal of Chromatography B1 (1994): 134-139.
  9. Raju NA., et al. “Estimation of Levetiracetam in tablet dosage form by RP-HPLC”. E-journal of Chemistry 5.S2 (2008): 1098-1102.
  10. Rao AL and Jahnavi V. “A validated RP-HPLC method for the estimation of Levetiracetam in bulk and pharmaceutical formulations”. E-Journal of Chemistry2 (2010): 600-604.
  11. Can NO and Goksel Arli. “Reversed-phase HPLC analysis of Levetiracetam in tablets using monolithic and conventional C18 silica columns”. Journal of AOAC International 4 (2010): 1077-1085.
  12. Prafulla Kumar Sahu., et al. “Development and validation of an RP-HPLC method for determination of Levetiracetamin bulk and pharmaceutical dosage forms”. Analytical Chemistry, An Indian Journal1 (2009): 34-38.
  13. Parimal Patel., et al. “Development and validation of stability-indicating UPLC method for estimation of Levetiracetam in bulk and pharmaceutical dosage form”. Journal of Pharmacy Research 12 (2011): 4495-4497.
  14. Mohammadi B., et al. “Simple and rapid ultra-high performance liquid chromatographic (UHPLC) method for the determination of Levetiracetam in human serum: Application to a bioequivalence study”. African Journal of Pharmacy and Pharmacology27 (2012): 2017-2022.
  15. Bhattacharya., et al. “Development of a validated stability-indicating high-performance thin-layer chromatographic method for the quantification of Levetiracetam”. Journal of Planar Chromatography2 (2014): 132-139.
  16. Jain DS., et al. “Determination of Levetiracetam in human plasma by liquid chromatography/electrospray tandem mass spectrometry and its application to bioequivalence studies”. Rapid Communications in Mass Spectrometry17 (2006): 2539-2547.
  17. Jenjirattithigarn N., et al. “Determination of plasma Levetiracetam level by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS-MS) and its application in pharmacokinetics studies in neonates”. Journal of Chromatography B 1085 (2018): 13-20.
  18. Blonk MI., et al. “Quantification of Levetiracetam in plasma of neonates by ultra-performance liquid chromatography-tandem mass spectrometry”. Journal of Chromatography B7-8 (2010): 675-681.

Copyright: © 2021 Malla Krishna Prasad and Mukthinuthalapati Mathrusri Annapurna . This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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