Development and Validation of a Effective and Reliable HPLC Method for the Quantification of Levodopa and Carbidopa in Pharmaceutical Formulations

Year 2021, Issue: 4, 413 - 422, 25.05.2021

İbrahim Bulduk , Süleyman Gökce

https://doi.org/10.15671/hjbc.795609

Cited By: 1

Abstract

A convenient and simple high performance liquid chromatography method for simultaneous separation, determination and identification of levodopa and carbidopa in pharmaceutical formulation has been developed. The analysis was carried out using Ace C18 (4.6 x 250 mm, 5 µm) column, and the separation was performed using a mobile phase consisting of 50 mM KH2PO4 (pH 2.3) pumped at a flow rate of 1.2 mL min-1 with UV detection at 280 nm. The method has been successfully validated on the basis of the International Conference on Harmonization (ICH) acceptance criteria. The method is selective, since no interferences corresponding to these analytes were found at retention times. Retention times for both drugs were 2.939 min and 5.018 for levodopa and carbidopa, respectively. The method was validated and shown to be linear in the concentration range of 25-125 µg mL-1 and 6.25-31.25 µg mL-1 for levodopa and carbidopa, respectively. The method exhibited good linearity (R2 > 0.999) over the assayed concentration range and showed good intra-day and inter-day precision. The limit of detection (LOD) and limit of quantification (LOQ) were determined based on standard deviation of the intercept and the slope of the calibration curve. LOD and LOQ values were 0.70 µg mL-1 and 2.13 µg mL-1 for levodopa and 1.31 µg mL-1 and 3.96 µg mL-1 for carbidopa, respectively. The method's accuracy was tested by adding known volume of standard solution (75 percent, 100 percent, and 125 percent concentration of the sample) to the 100 percent concentration pre-analyzed sample solution. The percentage mean recovery by standard addition experiments of levodopa and carbidopa is 99.75% and 99.55%, respectively. This method has also been successfully used for the determination of levodopa and carbidopa in pharmaceutical formulations.

Keywords

levodopa, carbidopa, validation, HPLC-UV

References

• 1. S. Fahn, D. Oakes, I. Shoulson, K.Kieburtz, A. Rudolph, A. Lang, CW. Olanow, C. Tanner, K. Marek, Levodopa and the progression of Parkinson's disease, The New England Journal of Medicine, 351 (2004) 2498–2508.
• 2. CE. Clarke, M. Guttman, Dopamine Agonist Monotherapy in Parkinson's Disease, Lancet, 360 (9347 (2002) 1767-9.
• 3. M. Rezak, Current pharmacotherapeutic treatment options in Parkinson's disease, Disease- a-Month: DM., 53 (2007) 214–222.
• 4. https://www.drugbank.ca/drugs/DB01235 Drug created on June 13, 2005 07:24 / Updated on June 12, 2020 11:41
• 5. D. Goncalves, G. Alves, P. Soares-da-Silva, A. Falcão, Bioanalytical chromatographic methods for the determination of catechol-O-methyltransferase inhibitors in rodents and human samples: a review, Analytica Chimica Acta, 710 (2012) 17–32.
• 6. https://www.drugbank.ca/drugs/DB00190 Drug created on June 13, 2005 07:24 / Updated on September 03, 2020 19:01
• 7. FJ. David, MR. Rafferty, JA. Robichaud, J. Prodoehl, WM. Kohrt, DE. Vaillan-court, DM. Corcos, Progressive resistance exercise and Parkinson's disease: a review of potential mechanisms. Parkinson’s Disease, (2011; 2012): 124527.
• 8. J.M. Cedarbaum, R.Williamson, H. Kutt, Simultaneous determination of levodopa, its metabolites and carbidopa in clinical samples, Journal of Chromatography B: Biomedical Sciences and Applications, 415 (1987) 393–399.
• 9. Y. Michotte, M. Moors, D.Deleu, P. Herregodts, G. Ebinger, 3-O-methyldopa and dopamine in plasma using high-performance liquid chromatography with electrochemical detection, Journal of Pharmaceutical and Biomedical Analysis, 5 (1987) 659–664.
• 10. DC. Titus, TF. August, KC. Yeh, R. Eisenhandler, WF. Bayne, DG. Musson, Simultaneous high-performance liquid chromatographic analysis of carbidopa, levodopa and 3-O-methyldopa in plasma and carbidopa, levodopa and dopamine in urine using electrochemical detection, Journal of Chromatography B: Biomedical Sciences and Applications, 534 (1990) 87–100.
• 11. KA. Sagar, MR. Smyth, Simultaneous determination of levodopa, carbidopa and their metabolites in human plasma and urine samples using LC-EC, J. Pharm. Biomed. Anal., 22 (2000) 613–624.
• 12. V. Rizzo, M. Memmi, R. Moratti, G. Melzi d’Eril, E. Perucca, Concentrations of L-dopa in plasma and plasma ultrafiltrates, Journal of Pharmaceutical and Biomedical Analysis, 14 (1996) 1043–1046.
• 13. T. Wikberg, Simultaneous determination of levodopa, its main metabolites and carbidopa in plasma by liquid chromatography, Journal of Pharmaceutical and Biomedical Analysis, 9 (1991); 167–176.
• 14. M. Karimi, JL. Carl, S.Loftin, JS. Perlmutter, Modified high-performance liquid chromatography with electrochemical detection method for plasma measurement of levodopa, 3-O-methyldopa, dopamine, carbidopa and 3,4-dihydroxyphenyl acetic acid, Journal of chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 836(1-2) (2006); 120-123.
• 15. F. Bugamelli, C. Marcheselli, E. Barba, MA. Raggi, Determination of L‐dopa, carbidopa, 3‐O‐methyldopa and entacapone in human plasma by HPLC‐ED, Journal of Pharmaceutical and Biomedical Analysis, 54(3) (2011) 562–567.
• 16. M. Kuoppamaki, K. Korpela, R. Marttila, V. Kaasinen, P. Hartikainen, J. Lyytinen, S. Kaakkola, J. Hanninen , E. Loyttyniemi, M. Kailajarvi, P. Ruokoniemi, J. Ellmen, Comparison of pharmacokinetic profile of levodopa throughout the day between levodopa/carbidopa/entacapone and levodopa/carbidopa when administered four or five times daily, European Journal of Clinical Pharmacology, 65 (2009) 443–455.
• 17. C. César Ida, RM. Byrro, FF. Cardoso, IM. Mundim, S. Teixeira Lde, EP. da Silva, SA. Gomes, RR. Bonfim, GA. Pianetti, Simultaneous quantitation of levodopa and 3-Omethyldopa in human plasma by HPLC–ESI-MS/MS: Application for a pharmacokinetic study with a levodopa/benserazide formulation, Journal of Pharmaceutical and Biomedical Analysis, 56 (2011) 1094–1100.
• 18. HF. Martins, DP. Pinto, AV. Nascimento, MAS. Marques, FC. Amendoeira, Development of an HPLC/MS/MS methodology for determining 3-Omethyldopa in human plasma and its application in a bioequivalence study, Química Nova, 36 (2013) 171–176.
• 19. K. Igarashi, K. Hotta, F. Kasuya, K. Abe, S. Sakoda, Determination of cabergoline and L-dopa in human plasma using liquid chromatography-tandem mass spectrometry, Journal of Chromatography B, 792 (2003) 55–61.
• 20. RR. Gonzalez, RF. Fernandez, JL. Vidal, AG. Frenich, J. Perez, Development and validation of an ultra-high performance liquid chromatography-tandem mass-spectrometry (UHPLC-MS/MS) method for the simultaneous determination of neurotransmitters in rat brain samples, Journal of Neuroscience Methods, 198 (2011) 187–194.
• 21. PO. Cinto, ALRS. Souza, AC. Lima, MV. Chaud, MPD. Gremião, LC Evaluation of Intestinal Transport of Praziquantel. Chromatographia, 69 (2009) 213–217.
• 22. F. Gosetti, E. Mazzucco, M. Gennaro, E. Marengo, Simultaneous determination of sixteen underivatized biogenic amines in human urine by HPLC-MS/MS, Analytical and Bioanalytical Chemistry (ABC), 405 (2013) 907–916.
• 23. C. Muzzi, E. Bertocci, L. Terzuoli, B. Porcelli, I. Ciari, R. Pagani, R. Guerranti, Simultaneous Determination of Serum Concentrations of Levodopa, Dopamine, 3-O-Methyldopa and α-Methyldopa by HPLC, Biomedicine & Pharmacotherapy, 62 (2008) 253–258.
• 24. HX. Zhao, H. Mu, YH. Bai, H. Yu, YM. Hu, A rapid method for the determination of dopamine in porcine muscle by pre-column derivatization and HPLC with fluorescence detection, Journal of Pharmaceutical Analysis, 1 (2011) 208–212.